2024-03-28T22:35:39Zhttp://uvadoc.uva.es/oai/requestoai:uvadoc.uva.es:10324/60772021-06-23T09:51:28Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ganfornina Álvarez, María Dolores
Carmo, Sonia do
Lora, Jose M.
Torres Schumann, Sonia
Vogel, Marci
Allhorn, Maria
González, Constancio
Bastiani, Michael J.
Rassart, Eric
Sánchez Romero, Diego
2014-09-19T10:04:53Z
2014-09-19T10:04:53Z
2008
Aging Cell, 2008, vol. 7, p. 506-515
1474-9718
http://uvadoc.uva.es/handle/10324/6077
10.1111/j.1474-9726.2008.00395.x
506
515
Aging Cell
7
Many nervous system pathologies are associated with
increased levels of apolipoprotein D (ApoD), a lipocalin
also expressed during normal development and aging.
An ApoD homologous gene in
Drosophila
, Glial Lazarillo,
regulates resistance to stress, and neurodegeneration in the
aging brain. Here we study for the first time the protective
potential of ApoD in a vertebrate model organism. Loss
of mouse ApoD function increases the sensitivity to
oxidative stress and the levels of brain lipid peroxidation,
and impairs locomotor and learning abilities. Human
ApoD overexpression in the mouse brain produces opposite
effects, increasing survival and preventing the raise of
brain lipid peroxides after oxidant treatment. These
observations, together with its transcriptional up-regulation
in the brain upon oxidative insult, identify ApoD as an acute
response protein with a protective and therefore beneficial
function mediated by the control of peroxidated lipids.
Key words: learning, lipid peroxidation, lipocalin, locomotor
behavior, paraquat, oxidative stress.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Nervioso, sistema - Enfermedades
Lípidos
Apolipoprotein D is involved in the mechanisms regulating protection from oxidative stress
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60782021-06-23T09:51:29Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ganfornina Álvarez, María Dolores
Sánchez Romero, Diego
Pagano, Aldo
Tonachini, Laura
Descalzi-Cancedda, Fiorella
Martínez, Salvador
2014-09-19T10:30:48Z
2014-09-19T10:30:48Z
2005
Developmental Dynamics, 2005, vol. 232, p. 191-199
1058-8388
http://uvadoc.uva.es/handle/10324/6078
10.1002/dvdy.20193
191
199
Developmental Dynamics
232
The insect Lazarillo and the mammalian apolipoprotein D (ApoD) are orthologous members of the lipocalin
protein family. We report the cloning and embryonic expression of chicken ApoD, the first molecularly
characterized nonmammalian ApoD. We also report the ApoD expression in mouse during postnatal
development and some novel aspects of the expression of the paralogous lipocalin prostaglandin D-synthase
(PGDS) and discuss these results in view of the lipocalin family evolution in vertebrates. ApoD is expressed
in subsets of central nervous system (CNS) neurons and glia during late chicken embryogenesis. Contrary
to mouse ApoD, no expression appears in neural crest-derived cephalic mesenchyme and blood vessel
pericytes. Also, ApoD is expressed in developing chicken feathers. These expressions are corroborated by
quantitative reverse transcriptase-polymerase chain reaction profiles. ApoD is expressed during mouse
postnatal development in a subset of CNS neurons, astrocytes and oligodendrocytes, but also in meninges
and pericytes. Chicken PGDS is expressed in brain meninges and perivascular cells. Our results suggest
that the amniote last common ancestor expressed ApoD and PGDS in the brain during embryogenesis. ApoD
appears restricted to ectodermal derivatives, whereas PGDS is expressed by derivatives of the three germ
layers.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Nervioso, Sistema - Evolución
Embriología
Células clonales
Molecular Characterization and Developmental Expression Pattern of the Chicken Apolipoprotein D Gene: Implications for the Evolution of Vertebrate Lipocalins
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60792021-06-23T09:51:30Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ruiz, Mario
Sánchez Romero, Diego
Canal, Inmaculada
Acebes, Angel
Ganfornina Álvarez, María Dolores
2014-09-19T10:39:18Z
2014-09-19T10:39:18Z
2011
Experimental Gerontology, 2011, vol. 46, p. 579-589
0531-5565
http://uvadoc.uva.es/handle/10324/6079
10.1016/j.exger.2011.02.014
579
589
Experimental Gerontology
46
Apolipoprotein D (ApoD), a member of the Lipocalin family, is the gene most up-regulated with age in the
mammalian brain. Its expression strongly correlates with aging-associated neurodegenerative and metabolic
diseases. Two homologues of ApoD expressed in the Drosophila brain, Glial Lazarillo (GLaz) and Neural
Lazarillo (NLaz), are known to alter longevity in male flies. However, sex differences in the aging process have
not been explored so far for these genes. Here we demonstrate that NLaz alters lifespan in both sexes, but
unexpectedly the lack of GLaz influences longevity in a sex-specific way, reducing longevity in males but not
in females. While NLaz has metabolic functions similar to ApoD, the regulation of GLaz expression upon aging
is the closest to ApoD in the aging brain. A multivariate analysis of physiological parameters relevant to
lifespan modulation uncovers both common and specialized functions for the two Lipocalins, and reveals that
changes in protein homeostasis account for the observed sex-specific patterns of longevity. The response to
oxidative stress and accumulation of lipid peroxides are among their common functions, while the
transcriptional and behavioral response to starvation, the pattern of daily locomotor activity, storage of fat
along aging, fertility, and courtship behavior differentiate NLaz from GLaz mutants. We also demonstrate that
food composition is an important environmental parameter influencing stress resistance and reproductive
phenotypes of both Lipocalin mutants. Since ApoD shares many properties with the common ancestor of
invertebrate Lipocalins, we must benefit from this global comparison with both GLaz and NLaz to understand
the complex functions of ApoD in mammalian aging and neurodegeneration.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Lipocainas
Células - Envejecimiento
Sex-dependent modulation of longevity by two Drosophila homologues of human Apolipoprotein D, GLaz and NLaz
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60812021-06-23T09:51:32Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Pagano, Aldo
Giannoni, Paolo
Zambotti, Adriana
Sánchez Romero, Diego
Ganfornina Álvarez, María Dolores
Gutiérrez, Gabriel
Randazzo, Nadia
Cancedda, Ranieri
Dozin, Beatrice
2014-09-19T10:48:54Z
2014-09-19T10:48:54Z
2004
Gene, 2004, vol. 331, p. 95-106
0378-1119
http://uvadoc.uva.es/handle/10324/6081
10.1016/j.gene.2004.02.001
95
106
Gene
331
A novel lipocalin gene is here reported that represents the fourth member of a cluster we have identified in the chicken genome. This
cluster also includes Chondrogenesis-Associated Lipocalins h and g (CALh, CALg) and Extracellular Fatty Acid Binding Protein (Ex-
FABP). The new gene codes for a 22-kDa secreted protein with three cysteine residues and a series of sequence features well conserved in the
lipocalin family. All the genes in the cluster are structurally similar presenting comparable exon/intron boundary positions and exon sizes. A
phylogenetic analysis indicates the monophyletic grouping of these genes, and their relationship with the lipocalins a-1-microglobulin
(A1mg), complement factor 8g chain (C8GC), prostaglandin D synthase (PGDS), and neutrophil-gelatinase-associated lipocalin (NGAL).
The new cluster gene appears to be the ortholog of the mammalian C8GC and was thus named Ggal-C8GC. This orthology also suggests that
this lipocalin was present in the ancestor common to reptiles and mammals.
In addition to other expressing tissues, Ex-FABP, CALh and CALg genes are highly transcribed in chondrocytes at late stages of
chondrogenesis during endochondral bone formation and/or upon inflammatory stimulation. Here, we show that they are also
transcriptionally induced when chondrocytes are subjected to various biological events as cell quiescence, cell shape transition, and hormonal
stimulation. By contrast, Ggal-C8GC transcripts are only barely detectable in chondrocytes, but are more abundant in liver, kidney, brain,
heart, skeletal muscle and particularly in skin. Moreover, no expression induction was observed neither during chondrocyte differentiation,
nor upon any of the stimulations mentioned above. This indicates that the Ggal-C8GC gene was co-opted for a novel function after the
duplication events that gave rise to the cluster.
The peculiar coordinated regulation of Ex-FABP, CALh and CALg, and the apparent divergent role of Ggal-C8GC suggest that these
gene duplications may have been maintained during evolution by a sub-functionalization mechanism where some common function(s) are
shared by several members of the cluster and some other specialized function(s) are unique to other members.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Lipocainas
Genética
Proteinas
Phylogeny and regulation of four lipocalin genes clustered in the chicken genome: evidence of a functional diversification after gene duplication
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60822021-06-23T09:51:34Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Suzuki, Kichiya
Lareyre, Jean-Jacques
Sánchez Romero, Diego
Gutiérrez, Gabriel
Araki, Yoshihiko
Matusik, Robert J.
Orgebin Crist, Marie Claire
2014-09-19T10:58:37Z
2014-09-19T10:58:37Z
2004
Gene, 2004, vol. 339, p. 49-59
0378-1119
http://uvadoc.uva.es/handle/10324/6082
10.1016/j.gene.2004.06.027
49
59
Gene
339
We previously identified two murine secretory proteins, mE-RABP(Lcn5) and mEP17(Lcn8), belonging to the lipocalin family and
specifically expressed in the epididymis. The genes are contiguous and localized on mouse chromosome 2. We now show that five other
related lipocalin genes, Lcn9, Lcn10, Lcn11, Lcn12, and Lcn13, that evolved by in situ tandem duplication are present on the same locus.
Lcn9, Lcn10, Lcn12, and Lcn13 genes, like Lcn5 and Lcn8 genes, are specifically expressed in the mouse epididymis. However, each gene
has a distinct spatial expression within the epididymis and different regulation. Analysis of the human genome sequence shows the presence
of genes encoding lipocalins with genomic organization, chromosomal arrangement, and orientation similar to that of the corresponding
murine genes, indicating that the epididymal cluster is evolutionary conserved. A phylogenetic analysis of the new epididymal proteins
reveals their spread position in the lipocalin protein family tree. This suggests the preservation of the regulatory sequences, while protein
sequences have greatly diverged, reflecting functional diversity and possibly multifunctionality. In terms of the cluster ancestry, epididymal
expression possibly appeared in a PGDS-like lipocalin in amniotes, and the duplications generating the cluster occurred at least in the
common ancestor of rodents and primates. The presence and conservation of a cluster of five genes encoding epididymal lipocalins,
differently regulated and regionalized in the epididymis, strongly suggests that these proteins may play an important role for male fertility.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Lipocainas
Genética
Fecundidad femenina
Molecular evolution of epididymal lipocalin genes localized on mouse chromosome 2
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60842021-06-23T09:51:36Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ganfornina Álvarez, María Dolores
Carmo, Sonia do
Martínez, Eva
Tolivia Fernández, Jorge Luis
Navarro, Ana
Rassart, Eric
Sánchez Romero, Diego
2014-09-19T11:09:58Z
2015-09-19T23:40:08Z
2010
Glia, 2010, vol. 58, p. 1320-1334
0894-1491
http://uvadoc.uva.es/handle/10324/6084
10.1002/glia.21010
1320
1334
Glia
58
Glial cells are a key element to the process of axonal regeneration,
either promoting or inhibiting axonal growth. The study
of glial derived factors induced by injury is important to
understand the processes that allow or preclude regeneration,
and can explain why the PNS has a remarkable ability to
regenerate, while the CNS does not. In this work we focus on
Apolipoprotein D (ApoD), a Lipocalin expressed by glial cells
in the PNS and CNS. ApoD expression is strongly induced
upon PNS injury, but its role has not been elucidated. Here we
show that ApoD is required for: (1) the maintenance of peripheral
nerve function and tissue homeostasis with age, and (2)
an adequate and timely response to injury. We study crushed
sciatic nerves at two ages using ApoD knock-out and transgenic
mice over-expressing human ApoD. The lack of ApoD
decreases motor nerve conduction velocity and the thickness
of myelin sheath in intact nerves. Following injury, we analyze
the functional recovery, the cellular processes, and the protein
and mRNA expression profiles of a group of injury-induced
genes. ApoD helps to recover locomotor function after injury,
promoting myelin clearance, and regulating the extent of
angiogenesis and the number of macrophages recruited to the
injury site. Axon regeneration and remyelination are delayed
without ApoD and stimulated by excess ApoD. The mRNA and
protein expression profiles reveal that ApoD is functionally
connected in an age-dependent manner to specific molecular
programs triggered by injury.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Células reproductoras
Lipocainas
ApoD, a Glia-Derived Apolipoprotein, Is Required for Peripheral Nerve Functional Integrity and a Timely Response to Injury
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60852021-06-23T09:51:38Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Bajo Grañeras, Raquel
Ganfornina Álvarez, María Dolores
Martín Tejedor, Esperanza
Sánchez Romero, Diego
2014-09-19T11:17:47Z
2015-09-19T23:40:08Z
2011
Glia, 2011, vol. 59, p. 1551-1566
0894-1491
http://uvadoc.uva.es/handle/10324/6085
10.1002/glia.21200
1551
1566
Glia
59
The study of glial derived factors induced by injury and
degeneration is important to understand the nervous system
response to deteriorating conditions. We focus on Apolipoprotein
D (ApoD), a Lipocalin expressed by glia and
strongly induced upon aging, injury or neurodegeneration.
Here we study ApoD function in the brain of wild type and
ApoD-KO mice by combining in vivo experiments with
astrocyte cultures. Locomotor performance, dopamine concentration,
and gene expression levels in the substantia
nigra were assayed in mice treated with paraquat (PQ). The
regulation of ApoD transcription, a molecular screening of
oxidative stress (OS)-related genes, cell viability and oxidation
status, and the effects of adding human ApoD were
tested in astrocyte cultures. We demonstrate that (1) ApoD
is required for an adequate locomotor performance, modifies
the gene expression profile of PQ-challenged nigrostriatal
system, and contributes to its functional maintenance; (2)
ApoD expression in astrocytes is controlled by the OSresponsive
JNK pathway; (3) ApoD contributes to an
autocrine protecting mechanism in astrocytes, avoiding peroxidated
lipids accumulation and altering the PQ transcriptional
response of genes involved in ROS managing and the
inflammatory response to OS; (4) Addition of human ApoD
to ApoD-KO astrocytes promotes survival through a mechanism
accompanied by protein internalization and modulation
of astroglial reactivity. Our data support that ApoD
contributes to the endurance of astrocytes and decreases
their reactivity level in vitro and in vivo. ApoD function as
a maintenance factor for astrocytes would suffice to explain
the observed protection by ApoD of OS-vulnerable dopaminergic
circuits in vivo.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Sistema nervioso central - Enfermedades
Apolipoprotein D mediates autocrine protection of astrocytes and controls their reactivity level, contributing to the functional maintenance of paraquat-challenged dopaminergic systems
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60882021-06-23T09:51:39Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Navarro, Juan A.
Ohmann, Elisabeth
Botella, Jose A.
Liebisch, Gerhard
Moltó, Maria D.
Ganfornina Álvarez, María Dolores
Schmitz, Gerd
Schneuwly, Stephan
Sánchez Romero, Diego
2014-09-19T11:34:39Z
2015-09-19T23:40:08Z
2010
Human Molecular Genetics, 2010, p. 1-13
0964-6906
http://uvadoc.uva.es/handle/10324/6088
10.1093/hmg/ddq183
1
13
Human Molecular Genetics
Friedreich’s ataxia (FRDA) is the most common form of autosomal recessive ataxia caused by a deficit in the
mitochondrial protein frataxin. Although demyelination is a common symptom in FRDA patients, no multicellular
model has yet been developed to study the involvement of glial cells in FRDA. Using the recently established
RNAi lines for targeted suppression of frataxin in Drosophila, we were able to study the effects of
general versus glial-specific frataxin downregulation. In particular, we wanted to study the interplay between
lowered frataxin content, lipid accumulation and peroxidation and the consequences of these effects on the
sensitivity to oxidative stress and fly fitness. Interestingly, ubiquitous frataxin reduction leads to an increase
in fatty acids catalyzing an enhancement of lipid peroxidation levels, elevating the intracellular toxic potential.
Specific loss of frataxin in glial cells triggers a similar phenotype which can be visualized by accumulating
lipid droplets in glial cells. This phenotype is associated with a reduced lifespan, an increased sensitivity
to oxidative insult, neurodegenerative effects and a serious impairment of locomotor activity. These symptoms
fit very well with our observation of an increase in intracellular toxicity by lipid peroxides.
Interestingly, co-expression of a Drosophila apolipoprotein D ortholog (glial lazarillo) has a strong protective
effect in our frataxin models, mainly by controlling the level of lipid peroxidation. Our results clearly support
a strong involvement of glial cells and lipid peroxidation in the generation of FRDA-like symptoms.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Células gliales
Altered lipid metabolism in a Drosophila model of Friedreich’s ataxia
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60922021-06-23T09:51:41Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ruiz, Mario
Wicker-Thomas, Claude
Sánchez Romero, Diego
Ganfornina Álvarez, María Dolores
2014-09-19T11:49:57Z
2014-09-19T11:49:57Z
2012
Insect Biochemistry and Molecular Biology, 2012, vol. 42, p. 776-789
0965-1748
http://uvadoc.uva.es/handle/10324/6092
10.1016/j.ibmb.2012.07.005
776
789
Insect Biochemistry and Molecular Biology
42
Lazarillo (Laz) is a glycosyl-phosphatidylinositol (GPI)-linked glycoprotein first characterized in the
developing nervous system of the grasshopper Schistocerca americana. It belongs to the Lipocalins,
a functionally diverse family of mostly secreted proteins. In this work we test whether the protective
capacity known for Laz homologs in flies and vertebrates (NLaz, GLaz and ApoD) is evolutionarily
conserved in grasshopper Laz, and can be exerted from the plasma membrane in a cell-autonomous
manner. First we demonstrate that extracellular forms of Laz have autocrine and paracrine protecting
effects for oxidative stress-challenged Drosophila S2 cells. Then we assay the effects of overexpressing
GPI-linked Laz in adult Drosophila and whether it rescues both known and novel phenotypes of NLaz null
mutants. Local effects of GPI-linked Laz inside and outside the nervous system promote survival upon
different stress forms, and extend lifespan and healthspan of the flies in a cell-type dependent manner.
Outside the nervous system, expression in fat body cells but not in hemocytes results in protection.
Within the nervous system, glial cell expression is more effective than neuronal expression. Laz actions
are sexually dimorphic in some expression domains. Fat storage promotion and not modifications in
hydrocarbon profiles or quantities explain the starvationedesiccation resistance caused by Laz overexpression.
This effect is exerted when Laz is expressed ubiquitously or in dopaminergic cells, but not in
hemocytes. Grasshopper Laz functionally restores the loss of NLaz, rescuing stress-sensitivity as well as
premature accumulation of aging-related damage, monitored by advanced glycation end products
(AGEs). However Laz does not rescue NLaz courtship behavioral defects. Finally, the presence of two new
Lipocalins with predicted GPI-anchors in mosquitoes shows that the functional advantages of
GPI-linkage have been commonly exploited by Lipocalins in the arthropodan lineage.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Lipocainas
Células gliales
Insectos
Grasshopper Lazarillo, a GPI-anchored Lipocalin, increases Drosophila longevity and stress resistance, and functionally replaces its secreted homolog NLaz
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60982021-06-23T09:51:43Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Bajo Grañeras, Raquel
Crespo Sanjuan, Jesús
García Centeno, Rosa María
Garrote Adrados, José Antonio
Gutiérrez, Gabriel
García Tejeiro, Manuel
Aguirre Cervás, Beatriz
Calvo Nieves, María Dolores
Bustamante, Rosa
Ganfornina Álvarez, María Dolores
Sánchez Romero, Diego
2014-09-19T16:17:35Z
2014-09-19T16:17:35Z
2013
International Journal of Colorectal Disease, 2013, p. 1-18
0179-1958
http://uvadoc.uva.es/handle/10324/6098
10.1007/s00384-012-1616-2
1
18
International Journal of Colorectal Disease
Purpose Inverse correlations of apolipoprotein D (ApoD)
expression with tumor growth have been shown, therefore
proposing ApoD as a good prognostic marker for diverse
cancer types, including colorectal cancer (CRC). Besides,
ApoD expression is boosted upon oxidative stress (OS) in
many pathological situations. This study aims at understanding
the role of ApoD in the progression of human CRC.
Methods Samples of CRC and distant normal tissue (n051)
were assayed for levels of lipid peroxidation, expression
profile of OS-dependent genes, and protein expression.
Three single-nucleotide polymorphisms in the ApoD gene
were analyzed (n0139), with no significant associations
found. Finally, we assayed the effect of ApoD in proliferation
and apoptosis in the CRC HT-29 cell line.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Colón - Cáncer - Tratamiento
Expression and potential role of apolipoprotein D on the death–survival balance of human colorectal cancer cells under oxidative stress conditions
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/60992021-06-23T09:51:45Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Bajo Grañeras, Raquel
Sánchez Romero, Diego
Gutiérrez, Gabriel
González, Constancio
Carmo, Sonia do
Rassart, Eric
Ganfornina Álvarez, María Dolores
2014-09-19T16:34:44Z
2015-09-19T23:40:08Z
2011
Journal of Neurochemistry, 2011, vol. 117, p. 949-960
0022-3042
http://uvadoc.uva.es/handle/10324/6099
10.1111/j.1471-4159.2011.07266.x
949
960
Journal of Neurochemistry
117
The lipocalin Apolipoprotein D (ApoD), known to protect the
nervous system against oxidative stress (OS) in model
organisms, is up-regulated early in the mouse brain in
response to the ROS generator paraquat. However, the processes
triggered by this up-regulation have not been explored.
We present here a study of the effect of ApoD on the early
transcriptional changes upon OS in the mouse cerebellum
using microarray profiling. ApoD-KO and transgenic mice
over-expressing ApoD in neurons are compared to wild-type
controls. In control conditions, ApoD affects the transcriptional
profile of neuron and oligodendrocyte-specific genes involved
in neuronal excitability, synaptic function, and myelin
homeostasis. When challenged with paraquat, the absence of
ApoD modifies the response of genes mainly related to OS
management and myelination. Interestingly, the over-expression
of ApoD in neurons almost completely abolishes the early
transcriptional response to OS. We independently evaluate
the expression of protein kinase Cd, a gene up-regulated by
OS only in the ApoD-KO cerebellum, and find it overexpressed
in cultured ApoD-KO primary astrocytes, which
points to a role for ApoD in astrocyte-microglia signaling. Our
results support the hypothesis that ApoD is necessary for a
proper response of the nervous system against physiological
and pathological OS.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Nervioso, sistema - Fisiología
Apolipoprotein D alters the early transcriptional response to oxidative stress in the adult cerebellum
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/61002021-06-18T11:04:58Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
Ganfornina Álvarez, María Dolores
Pérez García, María Teresa
Gutiérrez, Gabriel
Miguel Velado, Eduardo
López López, José Ramón
Marín, Antonio
Sánchez Romero, Diego
González, Constancio
2014-09-19T16:44:20Z
2015-09-19T23:40:08Z
2005
Journal of Physiology, 2005, vol. 566, n. 2, p. 491-503
0022-3751
http://uvadoc.uva.es/handle/10324/6100
10.1113/jphysiol.2005.088815
491
2
503
Journal of Physiology
566
The carotid body (CB) is an arterial chemoreceptor, bearing specialized type I cells that respond
to hypoxia by closing specific K+ channels and releasing neurotransmitters to activate sensory
axons. Despitehaving detailed informationonthe electricalandneurochemicalchangestriggered
by hypoxia in CB, the knowledge of the molecular components involved in the signalling cascade
of the hypoxic response is fragmentary. This study analyses the mouse CB transcriptional
changes in response to low PO2 by hybridization to oligonucleotide microarrays. The transcripts
were obtained from whole CBs after mice were exposed to either normoxia (21% O2),
or physiological hypoxia (10% O2) for 24 h. The CB transcriptional profiles obtained under
these environmental conditions were subtracted fromthe profile of control non-chemoreceptor
adrenal medulla extracted from the same animals. Given the common developmental origin of
these two organs, they share many properties but differ specifically in their response to O2. Our
analysis revealed 751 probe sets regulated specifically in CB under hypoxia (388 up-regulated
and 363 down-regulated). These results were corroborated by assessing the transcriptional
changesof selectedgenesunderphysiologicalhypoxiawithquantitativeRT-PCR.Ourmicroarray
experiments revealed a number of CB-expressed genes (e.g. TH, ferritin and triosephosphate
isomerase) that were known to change their expression under hypoxia. However, we also found
novel genes that consistently changed their expression under physiological hypoxia. Among
them, a group of ion channels show specific regulation in CB: the potassium channels Kir6.1 and
Kcnn4 are up-regulated, while the modulatory subunit Kcnab1 is down-regulated by low PO2
levels.
eng
info:eu-repo/semantics/embargoedAccess
Genética
Comparative gene expression profile of mouse carotid body and adrenal medulla under physiological hypoxia
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/61012021-06-23T09:51:46Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Jiménez Palomares, Margarita
Cózar Castellano, Irene
Ganfornina Álvarez, María Dolores
Sánchez Romero, Diego
Perdomo Hernández, Germán
2014-09-19T16:51:01Z
2014-09-19T16:51:01Z
2011
Metabolism, 2011, vol. 60, p. 1767-1774
0026-0495
http://uvadoc.uva.es/handle/10324/6101
10.1016/j.metabol.2011.04.013
1767
1774
Metabolism
60
Apolipoprotein D (ApoD) is an atypical apolipoprotein with an incompletely understood
function in the regulation of triglyceride and glucose metabolism. We have demonstrated
that elevated ApoD production in mice results in improved postprandial triglyceride
clearance. This work studies the role of ApoD deficiency in the regulation of triglyceride
and glucose metabolism and its dependence on aging. We used ApoD knockout (ApoD-KO)
mice of 3 and 21 months of age. Body weight and food intake were measured. Hepatic
histology, triglyceride content, lipoprotein lipase levels, and plasma metabolites were
studied. Phenotypic characterization of glucose metabolism was performed using glucose
tolerance test. β-Cell mass, islet volume, and islet number were analyzed by
histomorphometry. Apolipoprotein D deficiency results in nonfasting hypertriglyceridemia
in young (P = .01) and aged mice (P = .002). In young ApoD-KO mice, hypertriglyceridemiawas
associated with 30% to 50% increased food intake in nonfasting and fasting conditions,
respectively, without changes in body weight. In addition, lipoprotein lipase levels were
reduced by 35% in adipose tissue (P = .006). In aged ApoD-KO mice, hypertriglyceridemia was
not associated with changes in food intake or body weight, whereas hepatic triglyceride
levels were reduced by 35% (P = .02). Furthermore, nonfasting plasma insulin levels were
elevated by 2-fold in young (P = .016) and aged (P = .004) ApoD-KO mice, without changes in
blood glucose levels, glucose tolerance, β-cell mass, or islet number. These findings
underscore the importance of ApoD in the regulation of plasma insulin levels and
triglyceride metabolism, suggesting that ApoD plays an important role in the pathogenesis
of dyslipidemia.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Triglicéridos - Metabolismo
Glucosa - Metabolismo
Genetic deficiency of apolipoprotein D in the mouse is associated with nonfasting hypertriglyceridemia and hyperinsulinemia
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/61022021-06-23T09:51:47Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Sánchez Romero, Diego
Ganfornina Álvarez, María Dolores
Gutiérrez, Gabriel
Marín, Antonio
2014-09-19T17:00:48Z
2015-09-19T23:40:08Z
2003
Molecular Biology and Evolution, 2003, vol. 20, n. 5, p. 775-783
0737-4038
http://uvadoc.uva.es/handle/10324/6102
10.1093/molbev/msg079
775
5
783
Molecular Biology and Evolution
20
The Lipocalins are an ancient protein family whose expression is currently confirmed in bacteria, protoctists, plants,
arthropods, and chordates. The evolution of this protein family has been assessed previously using amino acid sequence
phylogenies. In this report we use an independent set of characters derived from the gene structure (exon-intron
arrangement) to infer a new lipocalin phylogeny. We also present the novel gene structure of three insect lipocalins. The
position and phase of introns are well preserved among lipocalin clades when mapped onto a protein sequence alignment,
suggesting the homologous nature of these introns. Because of this homology, we use the intron position and phase of 23
lipocalin genes to reconstruct a phylogeny by maximum parsimony and distance methods. These phylogenies are very
similar to the phylogenies derived from protein sequence. This result is confirmed by congruence analysis, and
a consensus tree shows the commonalities between the two source trees. Interestingly, the intron arrangement phylogeny
shows that metazoan lipocalins have more introns than other eukaryotic lipocalins, and that intron gains have occurred in
the C-termini of chordate lipocalins. We also analyze the relationship of intron arrangement and protein tertiary structure,
as well as the relationship of lipocalins with members of the proposed structural superfamily of calycins. Our congruence
analysis validates the gene structure data as a source of phylogenetic information and helps to further refine our
hypothesis on the evolutionary history of lipocalins.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Lipocainas
Calciproteinas
Exon-Intron Structure and Evolution of the Lipocalin Gene Family
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/61032021-06-23T09:51:48Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Boer, Simone
Sánchez Romero, Diego
Reinieren, Ivo
Boom, Tom van den
Udawela, Madhara
Scarr, Elizabeth
Ganfornina Álvarez, María Dolores
Dean, Brian
2014-09-19T17:07:39Z
2014-09-19T17:07:39Z
2010
Progress in Neuro-Psychopharmacology & Biological Psychiatry, 2010, vol. 34, p. 271-278
0278-5846
http://uvadoc.uva.es/handle/10324/6103
10.1016/j.pnpbp.2009.11.016
271
278
Progress in Neuro-Psychopharmacology & Biological Psychiatry
34
Apolipoprotein D (ApoD) has many actions critical to maintaining mammalian CNS function. It is therefore
significant that levels of ApoD have been shown to be altered in the CNS of subjects with schizophrenia,
suggesting a role for ApoD in the pathophysiology of the disorder. There is also a large body of evidence that
cortical and hippocampal glutamatergic, serotonergic and cholinergic systems are affected by the pathophysiology
of schizophrenia. Thus, we decided to use in vitro radioligand binding and autoradiography tomeasure levels
of ionotropic glutamate, somemuscarinic and serotonin 2Areceptors in theCNS ofApoD-/- and isogenic wild-type
mice. These studies revealed a 20% decrease(mean±SEM: 104±10.2 vs. 130±10.4 fmol/mg ETE) in the density
of kainate receptors in the CA 2–3 of the ApoD-/- mice. In addition therewas a global decrease inAMPA receptors
(F1,214=4.67, pb0.05) and a global increase in muscarinic M2/M4 receptors (F1,208=22.77, pb0.0001) in the
ApoD-/- mice that did not reach significance in any single cytoarchitectural region. We conclude that
glutamatergic pathways seem to be particularly affected in ApoD-/- mice and this may contribute to the changes
in learning and memory, motor tasks and orientation-based tasks observed in these animals, all of which involve
glutamatergic neurotransmission.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neuropsicofarmacología
Decreased kainate receptors in the hippocampus of apolipoprotein D knockout mice
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/61272021-06-23T09:51:50Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Hull-Thompson, Julie
Muffat, Julien
Sánchez Romero, Diego
Walker, David W.
Benzer, Seymour
Ganfornina Álvarez, María Dolores
Jasper, Heinrich
2014-09-22T15:46:39Z
2014-09-22T15:46:39Z
2009
PLoS Genetics, 2009, vol. 5, n. 4, p. 1-19
1553-7390
http://uvadoc.uva.es/handle/10324/6127
10.1371/journal.pgen.1000460
1
4
19
PLoS Genetics
5
Metabolic homeostasis in metazoans is regulated by endocrine control of insulin/IGF signaling (IIS) activity. Stress and
inflammatory signaling pathways—such as Jun-N-terminal Kinase (JNK) signaling—repress IIS, curtailing anabolic processes
to promote stress tolerance and extend lifespan. While this interaction constitutes an adaptive response that allows
managing energy resources under stress conditions, excessive JNK activity in adipose tissue of vertebrates has been found
to cause insulin resistance, promoting type II diabetes. Thus, the interaction between JNK and IIS has to be tightly regulated
to ensure proper metabolic adaptation to environmental challenges. Here, we identify a new regulatory mechanism by
which JNK influences metabolism systemically. We show that JNK signaling is required for metabolic homeostasis in flies
and that this function is mediated by the Drosophila Lipocalin family member Neural Lazarillo (NLaz), a homologue of
vertebrate Apolipoprotein D (ApoD) and Retinol Binding Protein 4 (RBP4). Lipocalins are emerging as central regulators of
peripheral insulin sensitivity and have been implicated in metabolic diseases. NLaz is transcriptionally regulated by JNK
signaling and is required for JNK-mediated stress and starvation tolerance. Loss of NLaz function reduces stress resistance
and lifespan, while its over-expression represses growth, promotes stress tolerance and extends lifespan—phenotypes that
are consistent with reduced IIS activity. Accordingly, we find that NLaz represses IIS activity in larvae and adult flies. Our
results show that JNK-NLaz signaling antagonizes IIS and is critical for metabolic adaptation of the organism to
environmental challenges. The JNK pathway and Lipocalins are structurally and functionally conserved, suggesting that
similar interactions represent an evolutionarily conserved system for the control of metabolic homeostasis.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Homeostasis
Lipocainas
Control of Metabolic Homeostasis by Stress Signaling Is Mediated by the Lipocalin NLaz
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/63392021-06-23T09:51:51Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
García García, Antonio
Padín, Fernando
Fernández Morales, Jose Carlos
Maroto, Marcos
García-Sancho Martín, Francisco Javier
2014-09-30T15:50:10Z
2014-09-30T15:50:10Z
2012
Cell Calcium, 2012, vol. 51, p. 309-320
0143-4160
http://uvadoc.uva.es/handle/10324/6339
10.1016/j.ceca.2011.12.004
309
320
Cell Calcium
51
The concept of stimulus–secretion coupling was born from experiments performed in chromaffin cells 50
years ago. Stimulation of these cells with acetylcholine enhances calcium (Ca2+) entry and this generates a
transient elevation of the cytosolic Ca2+ concentration ([Ca2+]c) that triggers the exocytotic release of catecholamines.
The control of the [Ca2+]c signal is complex and depends on various classes of plasmalemmal
calcium channels, cytosolic calcium buffers, the uptake and release of Ca2+ from cytoplasmic organelles,
such as the endoplasmic reticulum, mitochondria, chromaffin vesicles and the nucleus, and Ca2+ extrusion
mechanisms, such as the plasma membrane Ca2+-stimulated ATPase, and the Na+/Ca2+ exchanger.
Computation of the rates of Ca2+ fluxes between the different cell compartments support the proposal
that the chromaffin cell has developed functional calcium tetrads formed by calcium channels, cytosolic
calcium buffers, the endoplasmic reticulum, and mitochondria nearby the exocytotic plasmalemmal sites.
These tetrads shape the Ca2+ transients occurring during cell activation to regulate early and late steps of
exocytosis, and the ensuing endocytotic responses. The different patterns of catecholamine secretion in
response to stress may thus depend on such local [Ca2+]c transients occurring at different cell compartments,
and generated by redistribution and release of Ca2+ by cytoplasmic organelles. In this manner,
the calcium tetrads serve to couple the variable energy demands due to exo–endocytotic activities with
energy production and protein synthesis.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Calcio
Bioquímica
Cytosolic organelles shape calcium signals and exo–endocytotic responses of chromaffin cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/63412021-06-23T09:51:53Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
García-Sancho Martín, Francisco Javier
Diego, Antonio M. G. de
García, Antonio G.
2014-09-30T16:12:08Z
2014-09-30T16:12:08Z
2012
Pflügers Archiv European Journal of Physiology, 2012, vol. 464, p. 33-41
0031-6768
http://uvadoc.uva.es/handle/10324/6341
10.1007/s00424-012-1074-2
33
41
Pflügers Archiv European Journal of Physiology
464
Chromaffin cells are an excellent model for stimulus–
secretion coupling. Ca2+ entry through plasma membrane
voltage-operated Ca2+ channels (VOCC) is the trigger
for secretion, but the intracellular organelles contribute subtle
nuances to the Ca2+ signal. The endoplasmic reticulum
amplifies the cytosolic Ca2+ ([Ca2+]C) signal by Ca2+-
induced Ca2+ release (CICR) and helps generation of microdomains
with high [Ca2+]C (HCMD) at the subplasmalemmal
region. These HCMD induce exocytosis of the docked
secretory vesicles. Mitochondria close to VOCC take up
large amounts of Ca2+ from HCMD and stop progression
of the Ca2+ wave towards the cell core. On the other hand,
the increase of [Ca2+] at the mitochondrial matrix stimulates
respiration and tunes energy production to the increased
needs of the exocytic activity. At the end of stimulation,
[Ca2+]C decreases rapidly and mitochondria release the Ca2+
accumulated in the matrix through the Na+/Ca2+ exchanger.
VOCC, CICR sites and nearby mitochondria form functional
triads that co-localize at the subplasmalemmal area, where
secretory vesicles wait ready for exocytosis. These triads
optimize stimulus–secretion coupling while avoiding
propagation of the Ca2+ signal to the cell core. Perturbation
of their functioning in neurons may contribute to the genesis
of excitotoxicity, ageing mental retardation and/or neurodegenerative
disorders.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Nervioso, Sistema - Enfermedades
Fisiología
Mitochondria and chromaffin cell function
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/63442021-06-23T09:51:54Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
López Doménech, Guillermo
Serrat, Román
Mirra, Serena
D'Aniello, Salvatore
Somorjai, Ildiko
Abad, Alba
Vitureira, Nathalia
García Arumí, Elena
Alonso Alonso, María Teresa
Rodríguez Prados, Macarena
Burgaya, Ferrán
Andreu, Antoni L.
García-Sancho Martín, Francisco Javier
Trullas, Ramón
García Fernández, Jordi
Soriano, Eduardo
2014-09-30T16:39:25Z
2015-03-30T23:40:09Z
2012
Nature Communications, 2012, p. 1-12
2041-1723
http://uvadoc.uva.es/handle/10324/6344
10.1038/ncomms1829
1
12
Nature Communications
Brain function requires neuronal activity-dependent energy consumption. Neuronal energy supply is controlled by molecular mechanisms that regulate mitochondrial dynamics, including Kinesin motors and Mitofusins, Miro1-2 and Trak2 proteins. Here we show a new protein family that localizes to the mitochondria and controls mitochondrial dynamics. This family of proteins is encoded by an array of armadillo (Arm) repeat-containing genes located on the X chromosome. The Armcx cluster is unique to Eutherian mammals and evolved from a single ancestor gene (Armc10). We show that these genes are highly expressed in the developing and adult nervous system. Furthermore, we demonstrate that Armcx3 expression levels regulate mitochondrial dynamics and trafficking in neurons, and that Alex3 interacts with the Kinesin/Miro/Trak2 complex in a Ca2 + -dependent manner. Our data provide evidence of a new Eutherian-specific family of mitochondrial proteins that controls mitochondrial dynamics and indicate that this key process is differentially regulated in the brain of higher vertebrates.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Proteinas mitocondriales
Cerebro - Investigación
The Eutherian Armcx genes regulate mitochondrial trafficking in neurons and interact with Miro and Trak2
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/63462021-06-23T09:51:55Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Orozco, Lluis
Munar, Anna
Soler, Robert
Alberca Zaballos, Mercedes
Soler, Francesc
Huguet, Marina
Sentís, Joan
Sánchez García, Ana María de los Ángeles
García-Sancho Martín, Francisco Javier
2014-09-30T17:01:23Z
2015-09-30T23:40:08Z
2013
Transplantation, Junio 2013, vol. 95, n. 12, p. 1535-1541
0041-1337
http://uvadoc.uva.es/handle/10324/6346
10.1097/TP.0b013e318291a2da
1535
12
1541
Transplantation
95
Osteoarthritis is the most prevalent joint disease and a frequent cause of joint pain, functional loss, and
disability. Osteoarthritis often becomes chronic, and conventional treatments have demonstrated only modest clinical
benefits without lesion reversal. Cell-based therapies have shown encouraging results in both animal studies and a few
human case reports. We designed a pilot study to assess the feasibility and safety of osteoarthritis treatment with
mesenchymal stromal cells (MSCs) in humans and to obtain early efficacy information for this treatment.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Osteoartritis - Tratamiento
Treatment of Knee Osteoarthritis with Autologous Mesenchymal Stem Cells: A Pilot Study
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/63482021-06-23T09:51:57Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
García-Sancho Martín, Francisco Javier
2014-09-30T17:17:39Z
2015-09-30T23:40:08Z
2014
Journal of Physiology, 2014, p. 261-268
0022-3751
http://uvadoc.uva.es/handle/10324/6348
10.1113/jphysiol.2013.255661
261
268
Journal of Physiology
Cross-talk between organelles and plasma membrane Ca2+ channels is essential for
modulation of the cytosolic Ca2+ ([Ca2+]C) signals, but such modulation may differ among
cells. In chromaffin cells Ca2+ entry through voltage-operated channels induces calcium release
from the endoplasmic reticulum (ER) that amplifies the signal. [Ca2+]C microdomains as high
as 20–50 μM are sensed by subplasmalemmal mitochondria, which accumulate large amounts of
Ca2+ through the mitochondrial Ca2+ uniporter (MCU). Mitochondria confine the high-Ca2+
microdomains (HCMDs) to beneath the plasma membrane,where exocytosis of secretory vesicles
happens. Cell core [Ca2+]C is much smaller (1–2 μM). By acting as a Ca2+ sink, mitochondria
stabilise theHCMDin space and time. In non-excitableHEK293 cells, activation of store-operated
Ca2+ entry, triggered by ERCa2+ emptying, also generated subplasmalemmal HCMDs, but, in this
case, most of the Ca2+ was taken up by the ER rather than bymitochondria. The smaller size of the
[Ca2+]C peak in this case (about 2 μM)may contribute to this outcome, as the sarco-endoplasmic
reticulum Ca2+ ATPase has much higher Ca2+ affinity than MCU. It is also possible that the relative positioning of organelles, channels and effectors, as well as cytoskeleton and accessory
proteins plays an important role.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Calcio - Absorción
The coupling of plasma membrane calcium entry to calcium uptake by endoplasmic reticulum and mitochondria
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/63492021-06-23T09:51:59Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Orozco, Lluis
Munar, Anna
Soler, Robert
Alberca Zaballos, Mercedes
Soler, Francesc
Huguet, Marina
Sentís, Joan
Sánchez García, Ana María de los Ángeles
García-Sancho Martín, Francisco Javier
2014-09-30T17:38:47Z
2015-09-30T23:40:08Z
2014
Transplantation, Junio 2014, vol. 97, n. 11, p. 66-68
0041-1337
http://uvadoc.uva.es/handle/10324/6349
10.1097/TP.0000000000000167
66
11
68
Transplantation
97
Osteoarthritis is the most prevalent
joint disease and a frequent cause
of joint pain, functional loss, and disability
(1). Osteoarthritis often becomes
chronic, and conventional treatments
have demonstrated only modest clinical
benefits, without lesion reversal (2). Cellbased
therapies have shown encouraging
results in both animal studies and a
few human case reports.We have recently
published the results of a pilot clinical
trial designed to assess the feasibility
and safety of osteoarthritis treatment
with bone marrowYderived mesenchymal
stromal cells (MSCs) in 12 patients
with chronic knee pain unresponsive
to conservative treatments and radiologic
evidence of osteoarthritis (3). The
patients were treated with autologous
expanded bone marrow MSCs by intraarticular
injection (40 106 cells), and
clinical outcomes, including evaluations
of pain, disability, and quality of life, were
followed up for 1 year. Articular cartilage
quality was assessed by quantitative
magnetic resonance imaging (MRI) T2
mapping (3).
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Osteoartritis - Tratamiento
Treatment of Knee Osteoarthritis With Autologous Mesenchymal Stem Cells: Two-Year Follow-up Results
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/63502021-06-23T09:52:01Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Rodríguez García, Arancha
Rojo Ruiz, Jonathan
Navas Navarro, Paloma
Aulestia Araya, Francisco Javier
Gallego Sandin, Sonia
García-Sancho Martín, Francisco Javier
Alonso Alonso, María Teresa
2014-09-30T18:04:21Z
2014-09-30T18:04:21Z
2014
Proceedings of the National Academy of Sciences, Febrero 2014, vol. 111, n. 7, p. 2584-2589
1091-6490
http://uvadoc.uva.es/handle/10324/6350
10.1073/pnas.1316539111
2584
7
2589
Proceedings of the National Academy of Sciences
111
Genetically encoded calcium indicators allow monitoring subcellular
Ca2+ signals inside organelles. Most genetically encoded calcium
indicators are fusions of endogenous calcium-binding proteins
whose functionality in vivo may be perturbed by competition with
cellular partners.We describe here a novel family of fluorescent Ca2+
sensors based on the fusion of two Aequorea victoria proteins, GFP
and apo-aequorin (GAP). GAP exhibited a unique combination
of features: dual-excitation ratiometric imaging, high dynamic
range, good signal-to-noise ratio, insensitivity to pH and Mg2+,
tunable Ca2+ affinity, uncomplicated calibration, and targetability
to five distinct organelles. Moreover, transgenic mice for endoplasmic
reticulum-targeted GAP exhibited a robust long-term
expression that correlated well with its reproducible performance in
various neural tissues. This biosensor fills a gap in the actual repertoire
of Ca2+ indicators for organelles and becomes a valuable tool
for in vivo Ca2+ imaging applications.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Calcio
Radiología
GAP, an aequorin-based fluorescent indicator for imaging Ca2+ in organelles
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/66192021-06-23T09:50:56Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Obeso Cáceres, Ana María de la Luz
Rocher Martín, María Asunción
Sidone, S.
González, Constancio
2014-10-20T09:17:57Z
2014-10-20T09:17:57Z
1992
Neuroscience, vol.47, n.2. p.463-472
0306-4522
http://uvadoc.uva.es/handle/10324/6619
10.1016/0306-4522(92)90260-9
463
2
472
Neuroscience
47
Ahatraet-The present study utilized an in vitro preparation of the rabbit carotid body, with tissue
catecholamine stores labeled by incubation with ‘H-tyrosine. The goal was to characterize pharmacologically
the vol~g~~n&nt Ca*+ channels present in the type I (glomus) cells of this arterial chemoreceptor
organ, and to elucidate their role as pathways for Ca2+ entry. We found that release of ‘H-dopamine
induced by high external potassium was over 95% dependent on external cakium concentration and that
this release was 9&100% inhibited by the dihydropy~~ne antagonists, nisoldipine and nitrendipine, and
was potentiated by the dihydropyridine agonist, BayK 8444. Therefore, any stimulus-induced, cakiumdependent
release of 3H-dopamine that was inhibited by nisoldipine and potentiated by BayK 8644, was
considered to be supported by Ca2+ entry into the cells via voltage-dependent Ca2+ channels. Significant
differences were observed in the release of ‘H-dopamine induced by 75 vs 25mM K+. On prolonged
stimulation, release induced by 75 mM K+ was large and transient, whilst that induced by 25 mM K+,
although more moderate, was sustained. The release elicited by 75 mM K+ was inhibited approximately
90% by 1.5 mM Co2+ or 625 nM nisoldipine, while release by 25 mM K+ was completely blocked by
0.6 mM Co*+ or 125 nM nisoldipine. Low PO,-induced release of 3H-dopamine was 95% dependent on
Ca*+, and was inhibited by nisoldipine (625 nM) in a manner inversely proportional to the intensity of
hypoxic stimulation, i.e. 79% inhibition at a PO, of 49 Torr, and 20% inhibition at PO2 of 0 Torr. BayK
8644 potentiatcd the release induced by moderate hypoxic stimuli. Release elicited by high PCOJlow pH,
or by Na+-propionate or dinitrophenol~n~ining solutions, was approximately 80% Ca’+-dependent,
and the ~hyd~y~din~ failed to modify this release.
It is concluded that type I mlls possess vol~~de~nd~t Ca ‘+ channels sensitive to the dihydropy~dines,
which in agreement with previous el~trophysiolo~~l data should be defined as L-type Ca*+
channels. Calcium entry which supports the release of 3H-dopamine elicited by moderate hypoxia should
occur mainly through these channels while the release induced by strong hypoxic stimuli will be SetNed
by Ca2+ entry which occurs in part via voltage-dependent Ca2+ channels, and in part through an
additional pathway, probably a Na+/Ca2+ exchanger. The insensitivity to dihydropyridines of the release
of )H-dopamine induced by high 1DC02/low pH, Na+-propionate and dinitrophenol may indicate a
complete loss of efficacy of the drugs to modulate Ca 2+ channels under these conditions or more likely,
that other mechanisms are activated, probably the Na+-Ca’+ exchanger.
Carotid body (CB) chemoreceptors are thought to be
composite receptors in which the type I (glomus) cells
detect changes in blood PO,, PCO, and pH and
respond with the release of neurotransmitt~ to
activate the closely apposed chemosensory nerve
terminals.~** One such neurotransmitter that has
received considerable attention in recent years
and is known to be released by the type I cells is
dopamine (DA). This biogenic amine has been
shown to be released in proportion to both the
intensity of stimulation and the resultant sensory
discharge recorded from the carotid sinus nerve
$To whom correspondence should be addressed.
Abbr~~~~~~ CB, carotid body; CSN, carotid sinus nerve;
DA, dopamine; DHMA, dihydrox~~delic acid,
DOPAC, dihydroxyphenyl acetic acid; NE, norepinephrine.
(CSN). This relationship between stimulus
eng
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Cardiovascular, Aparato - Tratamiento
The role of dihydropydine-sensitive Ca2 + channels in stimulus-evoked catecholamine release from chemoreceptor cells of the carotid body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/67162021-06-23T09:50:58Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Gómez Niño, María Ángeles
Almaraz Gómez, Laura
González, Constancio
2014-10-24T07:07:01Z
2014-10-24T07:07:01Z
1992
Neuroscience Letters, 1992, vol. 140 p.1-4
0304-3940
http://uvadoc.uva.es/handle/10324/6716
10.1016/0304-3940(92)90667-v
1
4
Neuroscience Letters
140
Salicylates, at the high therapeutic doses used in the treatment of rheumatoid arthritis, produce an increase in ventilation and augment the carotid
body reactivity to hypoxic stimulus, leading to an exaggerated hyperventilation during hypoxia. These effects had been related to the action of
salicylates as uncouplers of oxidative phosphorylation. In the present study, carried out in an in vitro preparation of the rabbit carotid body, we show
that acetylsalicylic acid and indomethacin, two anti-inflammatory drugs that are also powerful inhibitors of cyclooxygenase, the prostaglandinsynthetizing
enzyme, produce an increase in the [3H]catecholamine release evoked by low oxygen stimulation. The drugs did not affect basal normoxic
release, a finding that suggests that at the concentration used these anti-inflammatory agents do not have uncoupling actions, and that their effects
on hypoxic-induced release of [3H]catecholamines is mediated by their specific action as cyclooxygenase inhibitors. In agreement with this suggestion
we found that prostaglandin E~ completely prevented the effects of both anti-inflammatory agents. In addition, our data indicate that endogenously
synt hetized prostaglandins are powerful modulators of chemoreceptor cell function.
eng
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Artritis reumatoide - Tratamiento
Potentiation by cyclooxygenase inhibitors of the release of catecholamines from the rabbit carotid body and its reversal by prostaglandin E2
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/67192021-06-23T09:51:14Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Dinger, Bruce
González, Constancio
Yoshizaki, Katsuaki
Fidone, Salvatore
2014-10-24T07:41:53Z
2014-10-24T07:41:53Z
1981
Neuroscience Letters, 1981, 21, p. 51-55
0304-3940
http://uvadoc.uva.es/handle/10324/6719
51
55
Neuroscience Letters
21
Specific dopamine receptors were studied in freshly dissected, unhomogenized rabbit carotid bodies
incubated in [3H]spiroperidol. Total binding and non-specific binding were determined in the absence
and presence of 0.2 #M (+)-butaclamol, respectively. Specific binding in normal carotid bodies
incubated at near saturating concentrations (0.38 nM) was 1.63 _+ 0.58 pmol/g of tissue. Chronic section
of the carotid sinus nerve (14 days) resulted in a 64070 reduction (P < 0.05) in specific binding. We
conclude that the majority of specific dopaminergic receptors are located on carotid sinus nerve afferent
terminals.
eng
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Cuerpo carótido
[3H]Spiroperidol binding in normal and denervated carotid bodies
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/67492021-06-23T09:51:15Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, Constancio
Fidone, Salvatore
2014-10-28T09:37:08Z
2014-10-28T09:37:08Z
1977
Neuroscience Letters, 6(1977) 95-99
0304-3940
http://uvadoc.uva.es/handle/10324/6749
95
99
Neuroscience Letters
6
Rabbit carotid bodies synthesized [3H]dopamine (DA) during a 3-h incubation
period in modified Tyrode's solution containing 40 uM [3H]tyrosine.
Following tbis loading period, the carotid bodies were exposed for one additional
hour to unlabelled Tyrode's solution equilibrated with either 10%
oxygen in nitrogen or with 100% oxygen. The carotid bodies exposed to low
O2 released 81% more [3H]dopamine during this one-hour period than the
carotid bodies exposed only to pure oxygen. These data suggest that hypoxia
induces release of DA from the carotid body.
eng
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Cuerpo carótido
Tiróides - Enfermedades
Increased release of [3H]dopamine during low O2 stimulation of rabbit carotid body in vitro
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68212021-12-01T10:19:24Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
López López, José Ramón
González, Constancio
2014-10-30T08:50:46Z
2014-10-30T08:50:46Z
1992
FEBS Letters, 1992, vol. 299, n. 3. p. 251-254
0014-5793
http://uvadoc.uva.es/handle/10324/6821
10.1016/0014-5793(92)80126-2
251
3
254
FEBS Letters
299
Carotid body (CB) chemoreceptors respond to a decrease
in arterial blood pOz with an increase in action
potential frequency in the sensory fibers innervating the
organ, the response developing in a few seconds [l].
Chemoreceptor cells of the CB are the Oz sensing structures,
releasing neurotransmitters in proportion to the
pOz decrease and producing an electrical discharge of
parallel intensity in the sensory nerve fibers of the carotid
sinus nerve [2,3]. Recently, it has been shown that
chemoreceptor cells possess an O,-sensitive K’ current
that is reversibly inhibited by decreasing pOz in the
bathing solution. The suggestion was made that this
inhibition can lead to cell depolarization and activation
of voltage-dependent Ca” channels [4,5]; the entry of
Ca”+ through these channels, that are dihydropyridinesensitive,
would trigger the release of neurotransmitters
[6,7]. To accept this sequence, the inhibition of the K’
current must precede the activation of the sensory nerve
discharges [8], that is, the inhibition mus: develop
within the initial 3 s after lowering pOz, the time elapsing
between a decrease in arterial pO1! and the increase
in the carotid sinus nerve action potential frequency [l].
On the other hand, the suggestion has been made that
the O?-sensor in chemoreceptor cells should be a hemoglobin-
like hemoprotein, but no direct evidence to
support this suggestion is available [l]. The presentstudy compares the time course of the inhibition of the
K’ current by low pO1 and that of Na” current inhibition
by TTX, which is known to occur in a few hundred
ms [SJ. I: is shown that the former inhibition is faster.
It is also shown that carbon monoxide, a very inert gas
that in biological systems only reacts with hemoproteins,
prevents the low PO,-induced inhibition of
chemorcceptor cells E;” current.
eng
info:eu-repo/semantics/openAccess
© 1992 Elsevier
Células - Receptores
Cuerpo carotídeo
Cells - Receptors
Carotic body
Time course of K+ current inhibition by low oxygen in chemoreceptor cells of adult rabbit carotid body. Effects of carbon monoxide
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68252021-06-23T09:51:16Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Obeso Cáceres, Ana María de la Luz
Almaraz Gómez, Laura
González, Constancio
2014-10-30T09:13:59Z
2014-10-30T09:13:59Z
1986
Brain Research 371(1986) 25-36
00006-8993
http://uvadoc.uva.es/handle/10324/6825
10.1016/0006-8993(86)90806-1
25
36
Brain Research
371
The process of chemosensory transduction in the arterial chemoreceptors is not well understood. According to the metabolic hypothesis
of chemoreception, a decrease in arterial pO 2 will produce a decrease in the adenosine triphosphate (ATP) content in the chcmosensory
type I cells, leading to release of a neurotransmitter and increased sensory neural activity. There is a paucity of direct experimental
support for this hypothesis, and in the present work, we have tested the postulates of the metabolic hypothesis in an in vitro
preparation of cat carotid body using 2-deoxy-D-glucose as an ATP-depleting agent. This preparation, when superfused with Tyrode
containing 5 mM Na+-pyruvate instead of glucose, responds normally to hypoxia, low pH and acetylcholinc, and maintains normal
ATP levels. Under these conditions, 2-deoxy-D-glucose is a chemostimulant, i.e. electrical activity in the carotid sinus nerve is increased,
with a threshold concentration of 0.25 mM and a maximum response at about 2-4 raM. These concentrations of 2-deoxyglucose
evoke a dose-dependent release of [3H]dopamine (synthesized from pH]tyrosine) from the carotid bodies which parallels the
electrical activity. The 2-deoxy-D-glucose-evoked release and electrical activity is dependent on the presence of extracellular Ca 2+ .
These same concentrations of 2-deoxy-D-glucose significantly reduce the ATP content of the carotid bodies. The events postulated bv
the metabolic hypothesis, i.e. decrease in ATP content, release of a putative neurotransminer and activation of the sensory nerve endings
are found to occur simultaneously. A possible cause-effect relationship between these three events is discussed.
eng
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Neurofisiología
Effects of 2-Deoxy-D-Glucose on In Vitro Cat Carotid Body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68422021-06-23T09:51:10Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Obeso Cáceres, Ana María de la Luz
Almaraz Gómez, Laura
González, Constancio
2014-10-30T12:17:57Z
2014-10-30T12:17:57Z
1985
Brain Research, 1985, vol. 348, p.64-68
0006-8993
http://uvadoc.uva.es/handle/10324/6842
10.1016/0006-8993(85)90360-9
64
68
Brain Research
348
An unsolved issue for the arterial chemoreceptors is the mechanism by which hypoxia and other natural stimuli lead to an increase of
activity in the carotid sinus nerve. According to the 'metabolic hypothesis', the hypoxic activation of the carotid body (CB) is mediated
by a decrease of the ATP levels in the type I cells, which then release a neurotransmitter capable of exciting the sensory nerve endings.
Using an in vitro preparation of cat CB, we report that ATP levels in the CB do in fact decrease when the organs are exposed to moderate,
short lasting hypoxia (5 min 20% 02). Additionally, we found that decreases in ATP levels induced by 2-deoxyglucose (2 mM) or
sodium cyanide (0.1 raM) are closely correlated with dopamine release from type I cells and electrical activity in the carotid sinus
nerve elicited by these agents. The possible cause-effect relationship of these events is discussed
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Correlation Between Adenosine Triphosphate Levels, Dopamine Release and Electrical Activity in the Carotid Body: Support for the Metabolic Hypothesis of Chemoreception
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68432021-06-23T09:51:18Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Dinger, Bruce
Almaraz Gómez, Laura
Hirano, T.
Yoshizaki, Katsuaki
González, Constancio
Gómez Niño, María Ángeles
Fidone, Salvatore
2014-10-30T12:34:34Z
2014-10-30T12:34:34Z
1991
Brain Research, 1991, vol.562, p.190-198
0006-8993
http://uvadoc.uva.es/handle/10324/6843
10.1016/0006-8993(91)90621-2
190
198
Brain Research
562
Acetylcholine and muscarinic agonists inhibit chemosensory activity in the rabbit carotid sinus nerve (CSN). Because the mechanism of
this inhibition is poorly understood, we have investigated the kinetics and distribution of muscarinic receptors in the rabbit carotid body with
the specific muscarinic antagonist [SH]quinuclidinylbenzitate ([3H]QNB). Equilibrium binding experiments identified displaceable binding sites
(1/~M atropine) with a K d = 71.46 pM and a Bm~ x = 9.23 pmol/g tissue. These binding parameters and the pharmacology of the displaceable
[SH]QNB binding sites are similar to specific muscannic receptors identified in numerous other nervous, muscular and glandular tissues.
Comparisons of specific binding in normal and chronic CSN-denervated carotid bodies suggest that musearinic receptors are absent on afferent
terminals in the carotid body; however, nearly 50% of the specific [3H]QNB binding is lost following chronic sympathectomy, suggesting
the presence of presynaptic muscarinic receptors on the sympathetic innervation supplying the carotid body vasculature. Autoradiographic
studies have localized the remainder of [3H]QNB binding sites to Iobules of type I and type II parenchymal cells. In separate
experiments, the muscarinic agonists, oxotremorine (100/~M) and bethanechol (100 ~tM) were shown to inhibit both the release of catecholamines
and the increased CSN activity evoked by nicotine (50/~M) stimulation of the in vitro carotid body, Our data suggest that muscarinic
inhibition in the rabbit carotid body is mediated by receptors located on type I cells which are able to modulate the excitatory actions
of acetylcholine at nicotinic sites.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Muscarinic receptor localization and function in rabbit carotid body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68442021-06-23T09:51:19Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Rigual Bonastre, Ricardo Jaime
González, E.
González, Constancio
Fidone, Salvatore
2014-10-30T13:01:23Z
2014-10-30T13:01:23Z
1986
Brain Research, 1986, vol. 374, p.101-109
0006-8993
http://uvadoc.uva.es/handle/10324/6844
10.1016/0006-8993(86)90398-7
101
109
Brain Research
374
The role of catecholamines (CAs) in cat carotid body chemoreception has been controversial. On the basis of pharmacological experiments,
it would appear that endogenous dopamine (DA) may act either as an inhibitory or excitatory transmitter. Neurochemical
studies on the effects of natural stimulation on the release of carotid body CAs in the cat have also been inconclusive. In the present
study, we have characterized the synthesis and release of CAs in the in vitro cat carotid body preparation in response to different levels
of hypoxic stimulation and have correlated these measures with the chemosensory activity of the carotid sinus nerve. The synthesis of
[3H]DA and [3H]norepinephrine was linear for at least 4 h in carotid bodies incubated with their natural precursor [~H]tyrosine. Synthesis
of both [3H]CAs plateaued when the [3H]tyrosine concentration in the media reached 40 uM, which is a concentration similar to
that found in cat plasma. Exposure of the animals to an atmosphere of 10% 02 in N~ for 3 h prior to removal and incubation of the carotid
bodies with [3H]tyrosine resulted in an approximately 100% increase in the rate of [3H]DA synthesis but no change in [3H]norepinephrine
synthesis. This selective increase in [3H]DA synthesis was not detected when [3H]dihydroxyphenylalanine was used as precursor.
Carotid bodies first incubated with [3H]tyrosine and later superfused with solutions equilibrated with different gas mixtures (0
100% 0 2 in N2) exhibited an increase in [3H]DA release and carotid sinus nerve discharge which were inversely related to the oxygen
concentration. This relationship was strongest for the weaker stimuli (between 50% and 20% O 2 in N2), where both nerve activity and
[3H]DA release increased almost in parallel. With lower oxygen concentrations (10% O 2 and 0% 02 equilibrated solutions), the increase
in the release of [3H]DA was proportionally greater than the increase in carotid sinus nerve discharge. Our results demonstrate
that hypoxic stimulation increases both the rate of synthesis and release of DA in the cat carotid body. Although the precise role of DA
in this chemoreceptor organ is presently unknown, our findings suggest that this biogenic amine plays a direct role in generating or
controlling the electrical activity in the carotid sinus nerve.
INTRODUCTION
The mammalian carotid body is an arterial chemoreceptor
organ activated by low paO2, low pH and
high p~CO2 (ref. 21). Structurally, the receptor complex
is formed by clusters of two types of cells, the
type I and type II cells; the clusters lie within a supporting
connective tissue matrix containing a dense
capillary net24, 41. Sensory fibers of the carotid sinus
nerve (CSN) penetrate these cell clusters to end in
synaptic apposition with type I cells, which are considered
to be preneural (receptor) elements. Two
perennial issues pertaining to arterial chemoreception,
concern first, whether the type 1 cells are in fact
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Synthesis and Release of Catecholamines by the Cat Carotid Body in Vitro: Effects of Hypoxic Stimulation
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68452021-06-23T09:52:02Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Dinger, Bruce
González, Constancio
Yoshizaki, Katsuaki
Fidone, Salvatore
2014-10-30T13:15:55Z
2014-10-30T13:15:55Z
1985
Brain Research, 1985, vol. 339, p. 295-304
0006-8993
http://uvadoc.uva.es/handle/10324/6845
295
304
Brain Research
339
Acetylcholine and nicotinic agents excite cat carotid body chemoreceptors and modify their response to natural stimuli. The present
experiments utilized [125I]a-bungarotoxin ([125I]a-BGT) to localize within the chemosensory tissue the possible sites of action of exogenous
and endogenous nicotinic cholinergic substances. In vitro equilibrium binding studies of intact carotid bodies determined a K d of
5.57 nM and a Bma x of 9.21 pmol/g of tissue. Chronic section (12-15 days) of the carotid sinus nerve (CSN) did not change the amount
of displaceable toxin binding. In contrast, the specific binding was reduced by 46% following removal of the superior cervical ganglion.
Light microscope autoradiography of normal, CSN-denervated and sympathectomized carotid bodies revealed displaceable binding
sites concentrated in lobules of type I and type II cells. Treatment of carotid bodies with 50 nM a-BGT in vitro reduced by 50% the
release of [3H]dopamine (synthesized from [3H]tyrosine) caused by hypoxia or nicotine, and also significantly reduced the stimulus-.
evoked discharges recorded from the CSN. The data suggest (1) an absence of ct-BGT binding sites on the afferent terminals of the
CSN and (2) that nicotinic receptors located within parenchymal cell lobules may modulate the release of catecholamines from these cells.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Localization and Function of Cat Carotid Body Nicotinic Receptors
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68472021-06-23T09:50:59Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Dinger, Bruce
González, Constancio
Yoshizaki, Katsuaki
Fidone, Salvatore
2014-10-30T16:12:19Z
2014-10-30T16:12:19Z
1981
Brain Research, 1981, n. 205. p. 187-193
0006-8993
http://uvadoc.uva.es/handle/10324/6847
187
205
193
Brain Research
The carotid body is an arterial chemosensory organ which detects changes in
blood gas tensions and pH, and reflexly contributes to the cardiorespiratory adjustments
which occur during hypoxia, hypercapnia and acidosis. However, the sensory
mechanisms involved in carotid chemoreception remain to be elucidated.
Morphologically, the carotid body consists of an association of elemental units,
or glomeruli, within a connective tissue stroma penetrated by a dense capillary net 5.
The glomeruli are comprised of catecholamine-rich type I, or chief cells, which are enveloped
by glial-like processes of type II, or sustentacular, cellsa,4,19. Sensory fibers
from the carotid sinus nerve penetrate the glomeruli to terminate in synaptic-like
apposition on type I cellst,18, 21.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Alpha-bungarotoxin binding in cat carotid body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68492021-06-23T09:50:57Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Gómez Niño, María Ángeles
Dinger, Bruce
González, Constancio
Fidone, Salvatore
2014-11-03T09:28:55Z
2014-11-03T09:28:55Z
1990
Brain Research 525(1990) 160-164
0006-8993
http://uvadoc.uva.es/handle/10324/6849
160
164
Brain Research
525
Recent studies suggest that preneural type I (glomus) cells in the arterial chemoreceptor tissue of the carotid body act as primary transducer
elements which respond to natural stimuli (low 02, pH or increased CO2) by releasing chemical transmitter agents capable of exciting the closely
apposed afferent nerve terminals. These type I cells contain multiple putative transmitters, but the identity of the natural excitatory agents
remains an unresolved problem in carotid body physiology. Characterization of putative transmitter involvement in the response to natural
and pharmacological stimuli has therefore become fundamental to further understanding of chemotransmission in this organ. The present study
demonstrates that a natural stimulus (hypoxia) evokes the release of dopamine (DA) and norepinephrine (NE) in approximate proportion to
their unequal stores in rabbit carotid body (DA release/NE release = 8.2). In contrast, nicotine (100/~M), a cholinornimetic agent thought
to act on the nicotinic receptors present on the type I cells, evokes the preferential release of NE (DA release/NE release = 0.17). These
findings suggest that distinct mechanisms are involved in a differential mobilization of these two cateeholamines from the rabbit carotid body.
eng
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http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Differential stimulus coupling to dopamine and norepinephrine stores in rabbit carotid body type I cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68512021-06-23T09:51:04Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Rigual Bonastre, Ricardo Jaime
González, E.
Fidone, Salvatore
González, Constancio
2014-11-03T11:59:43Z
2014-11-03T11:59:43Z
1984
Brain Resarch, 309(1984) 178-181
0006-8993
http://uvadoc.uva.es/handle/10324/6851
10.1016/0006-8993(84)91026-6
178
181
Brain Research
309
The rates of dopamine and noradrenaline synthesis in the cat carotid body (c.b.) are 5.9 _+ 0.58 pmol/c.b./2 h and 0.3 + 0.02
pmol/c.b./2 h, respectively. The synthesis is doubled when the organs are incubated at pH 7. Similarly, low pH induces a release of dopamine
from the c.b. which is proportional to increased activity in the carotid sinus nerve.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Effects of low pH on synthesis and release of catecholamines in the cat carotid body in vitro
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68522021-06-23T09:51:11Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Rocher Martín, María Asunción
Cáceres Pereira, Ana Isabel
Obeso Cáceres, Ana María de la Luz
González, Constancio
2014-11-03T12:50:15Z
2014-11-03T12:50:15Z
2011
Marine Drugs, 9(2011) 2683-2704
1660-3397
http://uvadoc.uva.es/handle/10324/6852
10.3390/md9122683
2683
2704
Marine Drugs
9
Abstract: Carotid bodies (CBs) are secondary sensory receptors in which the sensing
elements, chemoreceptor cells, are activated by decreases in arterial PO2 (hypoxic
hypoxia). Upon activation, chemoreceptor cells (also known as Type I and glomus cells)
increase their rate of release of neurotransmitters that drive the sensory activity in the
carotid sinus nerve (CSN) which ends in the brain stem where reflex responses are
coordinated. When challenged with hypoxic hypoxia, the physiopathologically most
relevant stimulus to the CBs, they are activated and initiate ventilatory and cardiocirculatory
reflexes. Reflex increase in minute volume ventilation promotes CO2 removal from alveoli
and a decrease in alveolar PCO2 ensues. Reduced alveolar PCO2 makes possible alveolar
and arterial PO2 to increase minimizing the intensity of hypoxia. The ventilatory effect, in
conjunction the cardiocirculatory components of the CB chemoreflex, tend to maintain an
adequate supply of oxygen to the tissues. The CB has been the focus of attention since the
discovery of its nature as a sensory organ by de Castro (1928) and the discovery of its
function as the origin of ventilatory reflexes by Heymans group (1930). A great deal of
effort has been focused on the study of the mechanisms involved in O2 detection. This
review is devoted to this topic, mechanisms of oxygen sensing. Starting from a summary of
the main theories evolving through the years, we will emphasize the nature and significance
of the findings obtained with veratridine and tetrodotoxin (TTX) in the genesis of current
models of O2-sensing.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Tetrodotoxin as a Tool to Elucidate Sensory Transduction Mechanisms: The Case for the Arterial Chemoreceptors of the Carotid Body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68902022-07-29T08:21:24Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Liu, X.
He, Le
Dinger, Bruce
González, Constancio
Stensaas, L.
Fidone, Salvatore
2014-11-04T09:39:49Z
2014-11-04T09:39:49Z
2011
Respiratry Physiology and Neurobiology : 2011, 178(3) 362-369
1569-9048
http://uvadoc.uva.es/handle/10324/6890
10.1016/j.resp.2011.03.006
362
178
369
Respiratory Physiology and Neurobiology
Experiments in recent years have revealed labile electrophysiological and neurochemical
phenotypes in primary afferent neurons exposed to specific stimulus conditions associated with
the development of chronic pain. These studies collectively demonstrate that the mechanisms
responsible for functional plasticity are primarily mediated by novel neuroimmune interactions
involving circulating and resident immune cells and their secretory products, which together
induce hyperexcitability in the primary sensory neurons. In another peripheral sensory modality,
namely the arterial chemoreceptors, sustained stimulation in the form of chronic hypoxia (CH)
elicits increased chemoafferent excitability from the mammalian carotid body. Previous studies
which focused on functional changes in oxygen-sensitive type I cells in this organ have only
partially elucidated the molecular and cellular mechanisms which initiate and control this adaptive
response. Recent studies in our laboratory indicate a unique role for the immune system in
regulating the chemo-adaptive response of the carotid body to physiologically relevant levels of
hypoxia.
eng
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http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Dolor crónico - Tratamiento
A chronic pain: inflammation-dependent chemoreceptor adaptation in rat carotid body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/68922021-06-23T09:51:21Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Davidsen, Peter K.
Herbert, Jhon M.
Antczak, Kim
Ferrer, Elisabet
Peinado, Víctor I.
González, Constancio
Roca, Josep
Egginton, Stuart
Barberá, Joan A.
Falciani, Francesco
2014-11-04T12:40:34Z
2014-11-04T12:40:34Z
2014
Genome Medicine 2014, 6:59
1756-994X
http://uvadoc.uva.es/handle/10324/6892
10.1186/s13073-014-0059-5
Genome Medicine
59
Background: A relatively large percentage of patients with chronic obstructive pulmonary disease (COPD) develop
systemic co-morbidities that affect prognosis, among which muscle wasting is particularly debilitating. Despite
significant research effort, the pathophysiology of this important extrapulmonary manifestation is still unclear. A key
question that remains unanswered is to what extent systemic inflammatory mediators might play a role in this
pathology.
Cigarette smoke (CS) is the main risk factor for developing COPD and therefore animal models chronically exposed
to CS have been proposed for mechanistic studies and biomarker discovery. Although mice have been successfully
used as a pre-clinical in vivo model to study the pulmonary effects of acute and chronic CS exposure, data suggest
that they may be inadequate models for studying the effects of CS on peripheral muscle function. In contrast,
recent findings indicate that the guinea pig model (Cavia porcellus) may better mimic muscle wasting.
Methods: We have used a systems biology approach to compare the transcriptional profile of hindlimb skeletal
muscles from a Guinea pig rodent model exposed to CS and/or chronic hypoxia to COPD patients with muscle
wasting.
Results: We show that guinea pigs exposed to long-term CS accurately reflect most of the transcriptional changes
observed in dysfunctional limb muscle of severe COPD patients when compared to matched controls. Using network
inference, we could then show that the expression profile in whole lung of genes encoding for soluble inflammatory
mediators is informative of the molecular state of skeletal muscles in the guinea pig smoking model. Finally, we show
that CXCL10 and CXCL9, two of the candidate systemic cytokines identified using this pre-clinical model, are indeed
detected at significantly higher levels in serum of COPD patients, and that their serum protein level is inversely
correlated with the expression of aerobic energy metabolism genes in skeletal muscle.
Conclusions: We conclude that CXCL10 and CXCL9 are promising candidate inflammatory signals linked to the
regulation of central metabolism genes in skeletal muscles. On a methodological level, our work also shows that a
system level analysis of animal models of diseases can be very effective to generate clinically relevant hypothesis.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
EPOC
A systems biology approach reveals a link between systemic cytokines and skeletal muscle energy metabolism in a rodent smoking model and human COPD
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/70312021-06-24T07:36:27Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
López López, José Ramón
González, Constancio
Ureña, J.
López Barneo, José
2014-11-07T12:13:03Z
2014-11-07T12:13:03Z
1989
Journal of General Phisiology, 1989, vol. 93, n. 5. p. 1001-1015
0022-1295
http://uvadoc.uva.es/handle/10324/7031
10.1085/jgp.93.5.1001
1001
1015
Journal of General Phisiology
93
The hypothesis that changes in environmental 02 tension (pOi) could
affect the ionic conductances of dissociated type I cells of the carotid body was
tested. Cells were subjected to whole-cell patch clamp and ionic currents were
recorded in a control solution with normal pO 2 (pO~ = 150 mmHg) and 3-5 min
after exposure to the same solution with a lower pO,. Na and Ca currents were
unaffected by lowering pO, to 10 mmHg, however, in all cells studied (n = 42)
exposure to hypoxia produced a reversible reduction of the K current. In 14 cells
exposed to a pO 2 of 10 mmHg peak K current amplitude decreased to 35 +_ 8% of
the control value. The effect of low pO2 was independent of the internal Ca 2+
concentration and was observed in the absence of internal exogenous nucleotides.
Inhibition of K channel activity by hypoxia is a graded phenomenon and in the
range between 70 and 120 mmHg, which includes normal pO, values in arterial
blood, it is directly correlated with pO 2 levels. Low pO2 appeared to slow down the
activation time course of the K current but deactivation kinetics seemed to be
unaltered. Type I cells subjected to current clamp generate large Na- and Cadependent
action potentials repetitively. Exposure to low pO~ produces a 4-10
mV increase in the action potential amplitude and a faster depolarization rate of
pacemaker potentials, which leads to an increase in the firing frequency. Repolarization
rate of individual action potentials is, however, unaffected, or slightly
increased. The selective inhibition of K channel activity by low pO, is a phenomenon
without precedents in the literature that explains the chemoreceptive properties
of type I cells. The nature of the interaction of molecular O, with the K channel
protein is unknown, however, it is argued that a hemoglobin-like O, sensor,
perhaps coupled to a G protein, could be involved.
eng
info:eu-repo/semantics/openAccess
© 1989 The Rockefeller University Press
Neurofisiología
Neurophysiology
Low pO2 selectively inhibits K channel activity in chemoreceptor cells of the mammalian carotid body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/70532021-06-23T09:51:01Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, Constancio
Conde, Silvia V.
Gallego Martín, Teresa
Olea Fraile, Elena
González Obeso, Elvira
Ramírez, María
Yubero Benito, Sara
Agapito Serrano, María Teresa
Gómez Niño, María Ángeles
Obeso Cáceres, Ana María de la Luz
Rigual Bonastre, Ricardo Jaime
Rocher Martín, María Asunción
2014-11-10T12:43:56Z
2014-11-10T12:43:56Z
2014
Frontiers in Neuroanatomy, Mayo 2014, vol 8
1662-5129
http://uvadoc.uva.es/handle/10324/7053
10.3389/fnana.2014.00025
Frontiers in Neuroanatomy
8
When de Castro entered the carotid body(CB)field,the organ was considered to be a small autonomic ganglion,a gland,a glomusorglomerulus,or a paraganglion. In his 1928 paper,de Castro concluded:“Insum,the Glomuscarotic umisinnervated by centripetal fibers,whose trophic center sare located in thesensory ganglia of the glossopharyngeal, and not by centrifugal[efferent] or secret o motor fibers a sisthe case for glands ; these are precisely the facts which lead to suppose that the Glomuscaroticumisa sensory organ.”A few pages down,de Castro wrote:“The Glomus represents an organ with multiplereceptors furnished with specialized receptor cells like those of the sensory organs [tastebuds?]...As aplausible hypothesis we propos et hattheGlomuscaroti cum represents a sensory organ, at present the only one in its kind, dedicated to capture certain qualitative variations in the composition of blood, a function that,possibly by are flex mechanism would have an effect on the functionalactivity of other organs... Therefore, thesensory fiber would not be directly stimulated by blood, but via the intermediation of the epithelial cell soft he organ, which, as their structures suggests, possess a secretory function which would participate in the stimulation of the centripetal fibers.”In our article we will recreat et he experiments that allowed Fernando de Castrotoreach this first conclusion. Also, we will scrutinize the natural endowment sand the scientific knowledge that drove de Castrotomaket the triple hypotheses : the CBaschemoreceptor (variationsinbloodcomposition),as a secondary sensory receptor which functioning involves a chemical synapse, and as a center, origin of systemicreflexes. After a brief account of the systemic reflex effects resulting from the CB stimulation, we wil lcomplete our article with a general view of the cellular-molecular mechanisms currently thought to be involved in the functionin go fthis arterial chemoreceptor.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Fernando de Castro and the discovery of the arterial chemoreceptors
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/70552021-06-23T09:51:12Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, Constancio
Kwok, Yan
Gibb, James
Fidone, Salvatore
2014-11-10T13:33:35Z
2014-11-10T13:33:35Z
1979
Brain Resarch, 172 (1972) 572-576
0006-8993
http://uvadoc.uva.es/handle/10324/7055
572
576
Brain Research
172
The carotid body is an arterial chemoreceptor organ responsive to blood levels
of pO2, pCOe and pH 13. The parenchymal tissue of the carotid body is composed
mainly of two cell types: the glomus or Type I cells, which are disposed together in
groups or glomeruli, and the sustentacular or Type II cells, which appear as glial-like
elements enclosing the glomeruli in capsular fashion 3,4. The Type I cells, which have
abundant dense-cored vesicles and are known to contain catecholaminesl, 2,11,15,
receive a sensory innervation from afferent fibers of the carotid sinus nerve 3. Recent
studies have also shown the presence of reciprocal synapses at these junctions between
afferent nerve terminals and Type I cells TM. In addition, these cells receive an efferent
innervation from both preganglionic and postganglionic sympathetic fibers which
reach the carotid body from the superior cervical ganglion 18
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Reciprocal modulation of tyrosine hydroxylasea activity in rat carotid body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/70912021-06-23T09:51:03Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Cayzac, A.
Rocher Martín, María Asunción
Obeso Cáceres, Ana María de la Luz
González, Constancio
Ricardi, D.
Kemp, P. K.
2014-11-12T12:46:00Z
2014-11-12T12:46:00Z
2011
Respiratory Physiology & Neurobiology 175 (2011) 80–89
1569-9048
http://uvadoc.uva.es/handle/10324/7091
10.1016/j.resp.2010.09.009
80
89
Respiratory Physiology & Neurobiology
175
An increase in intracellular Ca2+ is crucial to O2 sensing by the carotid body. Polyamines have been
reported to modulate both the extracellular Ca2+-sensing receptor (CaR) and voltage-gated Ca2+ channels
in a number of cell types. Using RT-PCR and immunohistochemistry, the predominant voltage-gated Ca2+
channels expressed in the adult rat carotid body were L (CaV1.2) and N (CaV2.2)-type. CaR mRNA could
not be amplified from carotid bodies, but the protein was expressed in the nerve endings. Spermine
inhibited the hypoxia-evoked catecholamine release from isolated carotid bodies and attenuated the
depolarization- and hypoxia-evoked Ca2+ influx into isolated glomus cells. In agreement with data from
carotid body, recombinant CaV1.2 was also inhibited by spermine. In contrast, the positive allosteric
modulator of CaR, R-568, was without effect on hypoxia-induced catecholamine release from carotid
bodies and depolarization-evoked Ca2+ influx into glomus cells. These data show that spermine exerts a
negative influence on carotid body O2 sensing by inhibiting L-type Ca2+ channels.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Respiración celular
Spermine attenuates carotid body glomus cell oxygen sensing by inhibiting
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/71212021-06-23T09:51:13Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, Constancio
Agapito Serrano, María Teresa
Rocher Martín, María Asunción
Martín González, María del Carmen
Vega Agapito, María Victoria
Gómez Niño, María Ángeles
Rigual Bonastre, Ricardo Jaime
Castañeda, Javier
Obeso Cáceres, Ana María de la Luz
2014-11-13T09:13:02Z
2014-11-13T09:13:02Z
2007
Respiratory Physiology & Neurobiology 157 (2007) 30–44
1569-9048
http://uvadoc.uva.es/handle/10324/7121
10.1016/j.resp.2007.01.016
30
44
Respiratory Physiology & Neurobiology
157
Superoxide anion is the most important reactive oxygen species (ROS) primarily generated in cells. The main cellular constituents with capabilities
to generate superoxide anion areNADPHoxidases and mitochondrial respiratory chain. The emphasis of our article is centered in critically examining
hypotheses proposing that ROS generated by NADPH oxidase and mitochondria are key elements in O2-sensing and hypoxic responses generation
in carotid body chemoreceptor cells. Available data indicate that chemoreceptor cells express a specific isoform of NADPH oxidase that is activated
by hypoxia; generated ROS acting as negative modulators of the carotid body (CB) hypoxic responses. Literature is also consistent in supporting
that poisoned respiratory chain can produce high amounts of ROS, making mitochondrial ROS potential triggers-modulators of the CB activation
elicited by mitochondrial venoms. However, most data favour the notion that levels of hypoxia, capable of strongly activating chemoreceptor cells,
would not increase the rate of ROS production in mitochondria, making mitochondrial ROS unlikely triggers of hypoxic responses in the CB.
Finally, we review recent literature on heme oxygenases from two perspectives, as potential O2-sensors in chemoreceptor cells and as generators
of bilirubin which is considered to be a ROS scavenger of major quantitative importance in mammalian cells.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Respiración celular
Chemoreception in the context of the general biology of ROS
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/71372021-06-23T09:51:23Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, Constancio
Agapito Serrano, María Teresa
Rocher Martín, María Asunción
Gómez Niño, María Ángeles
Rigual Bonastre, Ricardo Jaime
Castañeda, Javier
Conde, J. V.
Obeso Cáceres, Ana María de la Luz
2014-11-13T12:30:47Z
2014-11-13T12:30:47Z
2010
Respiratory Physiology & Neurobiology 174 (2010) 317–330
1569-9048
http://uvadoc.uva.es/handle/10324/7137
10.1016/j.resp.2010.09.002
317
330
Respiratory Physiology & Neurobiology
174
Oxygen-sensing and transduction in purposeful responses in cells and organisms is of great physiological
and medical interest. All animals, including humans, encounter in their lifespan many situations in which
oxygen availability might be insufficient, whether acutely or chronically, physiologically or pathologically.
Therefore to trace at the molecular level the sequence of events or steps connecting the oxygen
deficit with the cell responses is of interest in itself as an achievement of science. In addition, it is also of
great medical interest as such knowledge might facilitate the therapeutical approach to patients and to
design strategies to minimize hypoxic damage. In our article we define the concepts of sensors and transducers,
the steps of the hypoxic transduction cascade in the carotid body chemoreceptor cells and also
discuss current models of oxygen- sensing (bioenergetic, biosynthetic and conformational) with their
supportive and unsupportive data from updated literature. We envision oxygen-sensing in carotid body
chemoreceptor cells as a process initiated at the level of plasma membrane and performed by a hemoprotein,
which might be NOX4 or a hemoprotein not yet chemically identified. Upon oxygen-desaturation,
the sensor would experience conformational changes allosterically transmitted to oxygen regulated K+
channels, the initial effectors in the transduction cascade. A decrease in their opening probability would
produce cell depolarization, activation of voltage dependent calcium channels and release of neurotransmitters.
Neurotransmitters would activate the nerve endings of the carotid body sensory nerve to convey
the information of the hypoxic situation to the central nervous system that would command ventilation
to fight hypoxia.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
A revisit to O2 sensing and transduction in the carotid body chemoreceptors in the context of reactive oxygen species biology
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/71562021-06-23T09:51:08Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Dinger, Bruce
He, Le
Chen, J.
Liu, X.
González, Constancio
Sanders, K.
Hoidal, J.
Stensaas, L.
Fidone, Salvatore
Obeso Cáceres, Ana María de la Luz
2014-11-14T12:01:12Z
2014-11-14T12:01:12Z
2007
Respiratory Physiology & Neurobiology 157 (2007) 45–54
1569-9048
http://uvadoc.uva.es/handle/10324/7156
10.1016/j.resp.2006.12.003
45
54
Respiratory Physiology & Neurobiology
157
O2-sensing in the carotid body occurs in neuroectoderm-derived type I glomus cells where hypoxia elicits a complex chemotransduction cascade
involving membrane depolarization, Ca2+ entry and the release of excitatory neurotransmitters. Efforts to understand the exquisite O2-sensitivity of
these cells currently focus on the coupling between local PO2 and the open-closed state of K+-channels. Amongst multiple competing hypotheses
is the notion that K+-channel activity is mediated by a phagocytic-like multisubunit enzyme, NADPH oxidase, which produces reactive oxygen
species (ROS) in proportion to the prevailing PO2. In O2-sensitive cells of lung neuroepithelial bodies (NEB), multiple studies confirm that ROS
levels decrease in hypoxia, and that EM and K+-channel activity are indeed controlled by ROS produced by NADPH oxidase. However, recent
studies in our laboratories suggest that ROS generated by a non-phagocyte isoform of the oxidase are important contributors to chemotransduction,
but that their role in type I cells differs fundamentally from the mechanism utilized by NEB chemoreceptors. Data indicate that in response to
hypoxia, NADPH oxidase activity is increased in type I cells, and further, that increased ROS levels generated in response to low-O2 facilitate cell
repolarization via specific subsets of K+-channels.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiologia
The role of NADPH oxidase in carotid body arterial chemoreceptors
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/71572021-06-23T09:51:25Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Gómez Niño, María Ángeles
Agapito Serrano, María Teresa
Obeso Cáceres, Ana María de la Luz
González, Constancio
2014-11-14T12:13:32Z
2014-11-14T12:13:32Z
2009
Respiratory Physiology & Neurobiology 165 (2009) 104–111
1569-9048
http://uvadoc.uva.es/handle/10324/7157
10.1016/j.resp.2008.10.020
104
111
Respiratory Physiology & Neurobiology
165
Lowoxygen sensing in chemoreceptor cells involves the inhibition of specific plasma membrane K+ channels,
suggesting that mitochondria-derived reactive oxygen species (ROS) link hypoxia to K+ channel
inhibition, subsequent cell depolarization and activation of neurotransmitter release.We have used several
mitochondrial poisons, alone and in combination with the antioxidant N-acetylcysteine (NAC), and
quantify their capacity to alter GSH/GSSG levels and glutathione redox potential (EGSH) in rat diaphragm.
Selected concentrations of mitochondrial poisons with or without NAC were tested for their capacity to
activate neurotransmitter release in chemoreceptor cells and to alter ATP levels in intact rat carotid body
(CB).We found that rotenone (1 M), antimycin A (0.2 g/ml) and sodium azide (5mM) decreased EGSH;
NAC restored EGSH to control values. At those concentrations mitochondrial poisons activated neurotransmitter
release from CB chemoreceptor cells and decreased CB ATP levels, NAC being ineffective to modify
these responses. Additional experiments with 3-nitroprionate (5 mM), lower concentrations of rotenone
and dinitrophenol revealed variable relationships between EGSH and chemoreceptor cell neurotransmitter
release responses and ATP levels. These findings indicate a lack of correlation between mitochondrialgenerated
modifications of EGSH and chemoreceptor cells activity. This lack of correlation renders unlikely
that alteration of mitochondrial production of ROS is the physiological pathway chemoreceptor cells use
to signal hypoxia.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Effects of mitochondrial poisons on glutathione redox potential and carotid body chemoreceptor activity
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/71582021-06-23T09:51:26Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Obeso Cáceres, Ana María de la Luz
Gómez Niño, María Ángeles
Almaraz Gómez, Laura
Dinger, Bruce
Fidone, Salvatore
González, Constancio
2014-11-14T12:37:39Z
2014-11-14T12:37:39Z
1997
Brain Research 754 1997. 298–302
0006-8993
http://uvadoc.uva.es/handle/10324/7158
10.1016/s0006-8993(97)00185-6
298
302
Brain Research
754
Current concepts on the location and functional significance of nicotinic receptors in the carotid body rest on a-bungarotoxin binding
and autoradiographic studies. Using an in vitro preparation of the cat carotid body whose catecholamine deposits have been labeled by
prior incubation with the tritiated natural precursor w3Hxtyrosine, we have found that nicotine induces release of w3Hxcatecholamines in a
dose-dependent manner IC50s9.81 mM.. We also found that mecamylamine 50 mM. completely abolished the nicotine-induced
release, while a-bungarotoxin 100 nM; f20 times its binding Kd. only reduced the release by 56%. These findings indicate that
chemoreceptor cells, and perhaps other carotid body structures, contain nicotinic receptors that are not sensitive to a-bungarotoxin and
force a revision of the current concepts on cholinergic mechanisms in the carotid body chemoreception.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Receptores celulares
Evidence for two types of nicotinic receptors in the cat carotid body chemoreceptor cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/71682021-06-23T09:51:09Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, Constancio
Sanz Alfayate, Gloria
Agapito Serrano, María Teresa
Gómez Niño, María Ángeles
Rocher Martín, María Asunción
Obeso Cáceres, Ana María de la Luz
2014-11-17T08:45:18Z
2014-11-17T08:45:18Z
2002
Respiratory Physiology & Neurobiology 132 (2002) 17–41
1569-9048
http://uvadoc.uva.es/handle/10324/7168
10.1016/S1569-9048(02)00047-2
17
41
Respiratory Physiology & Neurobiology
132
Reactive oxygen species (ROS) are oxygen-containing molecular entities which are more potent and effective
oxidizing agents than is molecular oxygen itself. With the exception of phagocytic cells, where ROS play an important
physiological role in defense reactions, ROS have classically been considered undesirable byproducts of cell
metabolism, existing several cellular mechanisms aimed to dispose them. Recently, however, ROS have been
considered important intracellular signaling molecules, which may act as mediators or second messengers in many cell
functions. This is the proposed role for ROS in oxygen sensing in systems, such as carotid body chemoreceptor cells,
pulmonary artery smooth muscle cells, and erythropoietin-producing cells. These unique cells comprise essential parts
of homeostatic loops directed to maintain oxygen levels in multicellular organisms in situations of hypoxia. The
present article examines the possible significance of ROS in these three cell systems, and proposes a set of criteria that
ROS should satisfy for their consideration as mediators in hypoxic transduction cascades. In none of the three cell
types do ROS satisfy these criteria, and thus it appears that alternative mechanisms are responsible for the
transduction cascades linking hypoxia to the release of neurotransmitters in chemoreceptor cells, contraction in
pulmonary artery smooth muscle cells and erythropoietin secretion in erythropoietin producing cells.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Significance of ROS in oxygen sensing in cell systems with sensitivy to ohysiological hypoxia
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/71712021-06-23T09:51:27Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, Constancio
Sanz Alfayate, Gloria
Obeso Cáceres, Ana María de la Luz
Agapito Serrano, María Teresa
2014-11-17T13:07:37Z
2014-11-17T13:07:37Z
2004
Methods in enzymology, 2004, vol. 381, p.40-70
0076-6879/04
http://uvadoc.uva.es/handle/10324/7171
10.1016/S0076-6879(04)81003-6
40
70
Methods in enzymology
381
This article first presents some basic structural traits of the carotid body (CB) arterial chemoreceptors to understand the relationship between the arterial blood PO2 and the activation of chemoreceptor cells, which are the O2 sensing structures of the CB. Some considerations in relation to the intensity of CB blood flow and O2 consumption of the organ would allow us to define the threshold for the detection of the hypoxic stimulus, which would lead us to the cardinal theme of the article, namely whether at the PO2 levels detected by the CB there alterations in the genesis of re-active oxygen species (ROS). An alteration in the rate of ROS productionwould impinge on the glutathione system [reduced glutathione (GSH) and oxidized glutathione (GSSG)], causing modifications in the GSH/GSSG ratio that are detected by direct measurement; the GSH/GSSG system rep-resents the quantitatively most important mechanism to dispose ROS and to maintain the overall redox status or redox environment in mammalian cells.1 The relationship between GSH/GSSG and oxygen chemoreception is approached from two different points of view. We will measure GSH/GSSG levels and calculate the redox environment of the cells and correl-ation with the activity of chemoreceptor cells in normoxia and in hypoxia. We will also present data on pharmacological manipulation of the redox environment of the cells, as assessed by GSH/GSSG quotients, and pos-sible correlations with the level of activity of chemoreceptor cells. The possible mechanisms of coupling between ROS and the GSH/GSSG system to the cellular effector machineries have been reviewed.2,3
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
Role of Glutathione Redox State in Oxygen Sensing by Carotid Body Chemoreceptor Cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/167262021-06-23T09:52:05Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
López Acosta, José Francisco
Moreno Amador, José Luis
Jiménez Palomares, Margarita
Díaz Marrero, Ana R.
Cueto, Mercedes
Perdomo Hernández, Germán
Cózar Castellano, Irene
2016-04-11T08:15:07Z
2016-04-11T08:15:07Z
2013
PLoS ONE, (2013); 8(1): e52862
1932-6203
http://uvadoc.uva.es/handle/10324/16726
10.1371/journal.pone.0052862
e52862
1
PLoS One.
8
There is an urgency to find new treatments for the devastating epidemic of diabetes. Pancreatic β-cells viability and function are impaired in the two most common forms of diabetes, type 1 and type 2. Regeneration of pancreatic β-cells has been proposed as a potential therapy for diabetes. In a preliminary study, we screened a collection of marine products for β-cell proliferation. One unique compound (epoxypukalide) showed capability to induce β-cell replication in the cell line INS1 832/13 and in primary rat cell cultures. Epoxypukalide was used to study β-cell proliferation by [3H]thymidine incorporation and BrdU incorporation followed by BrdU/insulin staining in primary cultures of rat islets. AKT and ERK1/2 signalling pathways were analyzed. Cell cycle activators, cyclin D2 and cyclin E, were detected by western-blot. Apoptosis was studied by TUNEL and cleaved caspase 3. β-cell function was measured by glucose-stimulated insulin secretion. Epoxypukalide induced 2.5-fold increase in β-cell proliferation; this effect was mediated by activation of ERK1/2 signalling pathway and upregulation of the cell cycle activators, cyclin D2 and cyclin E. Interestingly, epoxypukalide showed protection from basal (40% lower versus control) and cytokine-induced apoptosis (80% lower versus control). Finally, epoxypukalide did not impair β-cell function when measured by glucose-stimulated insulin secretion. In conclusion, epoxypukalide induces β-cell proliferation and protects against basal and cytokine-mediated β-cell death in primary cultures of rat islets. These findings may be translated into new treatments for diabetes
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Diabetes
Células - Reproducción
Pancreas
Epoxypukalide induces proliferation and protects against cytokine-mediated apoptosis in primary cultures of pancreatic β-cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/167382021-06-23T09:52:07Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
López Acosta, José Francisco
Villa Pérez, Pablo
Fernández Díaz, Cristina María
Luis Román, Daniel Antonio de
Díaz Marrero, Ana R.
Cueto, Mercedes
Perdomo Hernández, Germán
Cózar Castellano, Irene
2016-04-11T11:45:57Z
2016-04-11T11:45:57Z
2015
Islets,(2015);7(2):e1078053
1938-2014
http://uvadoc.uva.es/handle/10324/16738
10.1080/19382014.2015.1078053
e1078053
2
Islets
7
Diabetes is a consequence of a decrease on functional β-cell mass. We have recently demonstrated that epoxypukalide (Epoxy) is a natural compound with beneficial effects on primary cultures of rat islets. In this study, we extend our previous investigations to test the hypothesis that Epoxy protects β-cells and improves glucose metabolism in STZ-induced diabetic mice. We used 3-months old male mice that were treated with Epoxy at 200 μg/kg body weight. Glucose intolerance was induced by multiple intraperitoneal low-doses of streptozotocin (STZ) on 5 consecutive days. Glucose homeostasis was evaluated measuring plasma insulin levels and glucose tolerance. Histomorphometry was used to quantify the number of pancreatic β-cells per islet. β-cell proliferation was assessed by BrdU incorporation, and apoptosis by TUNEL staining. Epoxy treatment significantly improved glucose tolerance and plasma insulin levels. These metabolic changes were associated with increased β-cell numbers, as a result of a two-fold increase in β-cell proliferation and a 50% decrease in β-cell death. Our results demonstrate that Epoxy improves whole-body glucose homeostasis by preventing pancreatic β-cell death due to STZ-induced toxicity in STZ-treated mice
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Diabetes - Tratamiento
Protective effects of epoxypukalide on pancreatic b-cells and glucose metabolism in STZ-induced diabetic mice
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/218092021-06-23T09:52:12Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Calvo Rodríguez, María
García Durillo, Mónica
Villalobos Jorge, Carlos
Núñez Llorente, Lucía
2016-12-16T12:17:52Z
2016-12-16T12:17:52Z
2016
Biochim Biophys Acta Mol Cel Res 1863(11): 2637-2649
1388-1981
http://uvadoc.uva.es/handle/10324/21809
10.1016/j.bbamcr.2016.08.001
Aging is associated to cognitive decline and susceptibility to neuron death, two processes related recently to subcellular
Ca2+ homeostasis. Memory storage relies on mushroom spines stability that depends on store-operated
Ca2+ entry (SOCE). In addition, Ca2+ transfer from endoplasmic reticulum(ER) to mitochondria sustains energy
production but mitochondrial Ca2+ overload promotes apoptosis. We have addressed whether SOCE and ERmitochondria
Ca2+ transfer are influenced by culture time in long-term cultures of rat hippocampal neurons, a
model of neuronal aging.We found that short-term cultured neurons show large SOCE, low Ca2+ store content
and no functional coupling between ER and mitochondria. In contrast, in long-term cultures reflecting aging neurons,
SOCE is essentially lost, Stim1 and Orai1 are downregulated, Ca2+ stores becomeoverloaded, Ca2+ release is
enhanced, expression of the mitochondrial Ca2+ uniporter (MCU) increases and most Ca2+ released from the ER
is transferred to mitochondria. These results suggest that neuronal aging is associated to increased ERmitochondrial
cross talking and loss of SOCE. This subcellular Ca2+ remodeling might contribute to cognitive decline
and susceptibility to neuron cell death in the elderly.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Hippocampal neurons
In vitro aging promotes endoplasmic reticulum (ER)-mitochondria Ca2+ cross talk and loss of store-operated Ca2+ entry (SOCE) in rat hippocampal neurons
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/218112021-06-23T09:52:15Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Calvo Rodríguez, María
García Durillo, Mónica
Villalobos Jorge, Carlos
Núñez Llorente, Lucía
2016-12-16T12:26:56Z
2016-12-16T12:26:56Z
2016
J Alzheimers Dis. 2016 Jul 22;54(1):207-21
1387-2877
http://uvadoc.uva.es/handle/10324/21811
10.3233/JAD-151189
The most important risk factor for Alzheimer’s disease (AD) is aging. Neurotoxicity in AD has been linked to
dyshomeostasis of intracellular Ca2+ induced by small aggregates of the amyloid- peptide 1-42 (A 42 oligomers). However,
how aging influences susceptibility to neurotoxicity induced by A 42 oligomers is unknown. In this study, we used longterm
cultures of rat hippocampal neurons, a model of neuronal in vitro aging, to investigate the contribution of aging to
Ca2+ dishomeostasis and neuron cell death induced by A 42 oligomers. In addition, we tested whether non-steroidal antiinflammatory
drugs (NSAIDs) and R-flurbiprofen prevent apoptosis acting on subcellular Ca2+ in aged neurons.We found that
A 42 oligomers have no effect on young hippocampal neurons cultured for 2 days in vitro (2 DIV). However, they promoted
apoptosis modestly in mature neurons (8 DIV) and these effects increased dramatically after 13 DIV, when neurons display
many hallmarks of in vivo aging. Consistently, cytosolic and mitochondrial Ca2+ responses induced by A 42 oligomers
increased dramatically with culture age. At low concentrations, NSAIDs and the enantiomer R-flurbiprofen lacking antiinflammatory
activity prevent Ca2+ overload and neuron cell death induced by A 42 oligomers in aged neurons. However, at
high concentrations R-flurbiprofen induces apoptosis. Thus, A 42 oligomers promote Ca2+ overload and neuron cell death
only in aged rat hippocampal neurons. These effects are prevented by low concentrations of NSAIDs and R-flurbiprofen
acting on mitochondrial Ca2+ overload.
eng
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http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Alzheimer, Enfermedad de
Aging Enables Ca2+ Overload and Apoptosis Induced by Amyloid- Oligomers in Rat Hippocampal Neurons: Neuroprotection by Non-Steroidal Anti-Inflammatory Drugs and R-Flurbiprofen in Aging Neurons
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/218122021-06-23T09:52:09Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Caballero, Erica
Calvo Rodríguez, María
Gonzalo Ruiz, Alicia
Villalobos Jorge, Carlos
Núñez Llorente, Lucía
2016-12-16T12:35:48Z
2016-12-16T12:35:48Z
2016
Neuroscience Letters Volume 612, 26 January 2016, Pages 66–73
0304-3940
http://uvadoc.uva.es/handle/10324/21812
10.1016/j.neulet.2015.11.041
Oligomers of the amyloid peptide (A o) are becoming the most likely neurotoxin in Alzheimer’s disease.
Controversy remains on the mechanisms involved in neurotoxicity induced by A o and the targets
involved. We have reported that A o promote Ca2+ entry, mitochondrial Ca2+ overload and apoptosis
in cultured cerebellar neurons. However, recent evidence suggests that some of these effects could be
induced by glutamate receptor agonists solved in F12, the media in which A o are prepared. Here we
have tested the effects of different media on A o formation and on cytosolic Ca2+ concentration ([Ca2+]cyt)
in rat cerebellar and hippocampal cell cultures. We found that A o prepared according to previous protocols
but solved in alternative media including saline, MEM and DMEM do not allow oligomer formation
and fail to increase [Ca2+]cyt. Changes in the oligomerization protocol and supplementation of media with
selected salts reported to favor oligomer formation enable A o formation. A o prepared by the new procedure
and containing small molecular weight oligomers increased [Ca2+]cyt, promoted mitochondrial
Ca2+ overload and cell death in cerebellar granule cells and hippocampal neurons. These results foster a
role for Ca2+ entry in neurotoxicity induced by A o and provide a reliable procedure for investigating the
Ca2+ entry pathway promoted by A o.
eng
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http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Alzheimer, Enfermedad de
A new procedure for amyloid oligomers preparation enables the unambiguous testing of their effects on cytosolic and mitochondrial Ca2+ entry and cell death in primary neurons
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/218192021-06-23T09:52:10Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Calvo Rodríguez, María
Villalobos Jorge, Carlos
Núñez Llorente, Lucía
2016-12-19T06:56:28Z
2018-01-01T00:40:22Z
2015
J Vis Exp. 2015 Dec 1;(106)
1940-087X
http://uvadoc.uva.es/handle/10324/21819
10.3791/53330
Susceptibility to neuron cell death associated to neurodegeneration and ischemia are exceedingly increased in the aged brain but mechanisms
responsible are badly known. Excitotoxicity, a process believed to contribute to neuron damage induced by both insults, is mediated by activation
of glutamate receptors that promotes Ca2+ influx and mitochondrial Ca2+ overload. A substantial change in intracellular Ca2+ homeostasis or
remodeling of intracellular Ca2+ homeostasis may favor neuron damage in old neurons. For investigating Ca2+ remodeling in aging we have
used live cell imaging in long-term cultures of rat hippocampal neurons that resemble in some aspects aged neurons in vivo. For this end,
hippocampal cells are, in first place, freshly dispersed from new born rat hippocampi and plated on poli-D-lysine coated, glass coverslips. Then
cultures are kept in controlled media for several days or several weeks for investigating young and old neurons, respectively. Second, cultured
neurons are loaded with fura2 and subjected to measurements of cytosolic Ca2+ concentration using digital fluorescence ratio imaging. Third,
cultured neurons are transfected with plasmids expressing a tandem of low-affinity aequorin and GFP targeted to mitochondria. After 24 h,
aequorin inside cells is reconstituted with coelenterazine and neurons are subjected to bioluminescence imaging for monitoring of mitochondrial
Ca2+ concentration. This three-step procedure allows the monitoring of cytosolic and mitochondrial Ca2+ responses to relevant stimuli as for
example the glutamate receptor agonist NMDA and compare whether these and other responses are influenced by aging. This procedure may
yield new insights as to how aging influence cytosolic and mitochondrial Ca2+ responses to selected stimuli as well as the testing of selected
drugs aimed at preventing neuron cell death in age-related diseases.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Hippocampal neurons
Fluorescence and Bioluminescence Imaging of Subcellular Ca2+ in Aged Hippocampal Neurons
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/218202021-06-23T09:52:14Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Calvo Rodríguez, María
Villalobos Jorge, Carlos
Núñez Llorente, Lucía
2016-12-19T07:05:28Z
2016-12-19T07:05:28Z
2015
Neural Regen Res 10 (9), 1371-1372.
1673-5374
http://uvadoc.uva.es/handle/10324/21820
10.4103/1673-5374.165219
The most important risk factor for stroke and neurodegeneration is
aging. In fact, survival after stroke diminishes largely with aging. In
fact, recovery after brain artery occlusion is dramatically worsened
by aging, even normal aging is associated with neuron damage and
cognitive decline. Mechanisms involved in aging-related, cognitive
decline and susceptibility to neuron damage in stroke and neurodegeneration
are largely unknown. One of the most important mechanisms
contributing to neural dysfunction and death is excitotoxicity.
This process is based on the fact that the excessive glutamate
receptor stimulation may lead to neuronal damage. This overstimulation
may be due to increased concentration of glutamate, or the
prolonged activation of receptors.
eng
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Attribution-NonCommercial-NoDerivatives 4.0 International
Mitochondria
Non-steroidal anti-inflammatory drugs (NSAIDs) and neuroprotection in the elderly: a view from the mitochondria
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/241092021-06-23T09:52:03Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González, J.L.
2017-07-04T09:16:26Z
2017-07-04T09:16:26Z
1987
Medicina Clínica, 1987, vol. 89, n. 3 p.130-131
http://uvadoc.uva.es/handle/10324/24109
130
3
131
Medicina Clínica
89
En 1970 la Organización Mundial de la Salud
(QMSi puso en marcha un programa internacional de notificación voluntaria de RAM en el que participa nuestro país, oficialmente desde 1983, con un Centro Nacional en Barcelona.
El sistema, cuya realización ventajas han sido detalladamente descntos,utiliza el mé
todo de " tarjeta amarilla., el cual, aunque con claras !imitaciones en comparación con los métodos de vigilancia intensiva". ha sido recornendao por la OMS también para las no tificaciones procedentes del hospital.
spa
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Universidad de Valladolid
Attribution-NonCommercial-NoDerivatives 4.0 International
Reacciones adversas a medicamentos como urgencias externas en el Hospital Clínico Universitario de Valladolid
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/247172021-12-01T10:05:57Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
López Barneo, José
López López, José Ramón
Ureña, J.
González, Constancio
2017-07-26T08:00:57Z
2017-07-26T08:00:57Z
1988
Science, vol. 241. p.580-582.
0036-8075
http://uvadoc.uva.es/handle/10324/24717
580
582
Science
241
The ionic currents of carotid body type 1cells and their possible involvement in the detection of oxygen tension (Po2) in arterial blood are unknown. The electrical properties of these cells were studied with the whole-cell patch clamp technique, and the hypothesis that ionic conductances can bealtered by changes in Po2 was tested. The results show that type 1cells have voltage-dependent sodium, calcium, and potassium channels. Sodium and calcium currents were unaffected by a decrease in Po2 from 150 to 10 millimeters of mercury, whereas, with the same experimental protocol, potassi um currents were reversibly reduced by 25 to 50 percent. The effect of hypoxia was independent of internal adenosine triphosphate and calcium. Thus, ionic conductances, and particularly the 02-sensitive potassium current, play a key role in the transduction mechanism of arterial chemoreceptors.
spa
info:eu-repo/semantics/openAccess
Chemotransduction in the carotid body: K+ current modulated by PO2 in type I chemoreceptor cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/247242021-06-24T07:36:25Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
Ureña, J.
López López, José Ramón
González, Constancio
López Barneo, José
2017-07-26T08:42:06Z
2017-07-26T08:42:06Z
1989
Journal of General Physiology, 1989, vol. 93. p. 979-999
0022-1295
http://uvadoc.uva.es/handle/10324/24724
10.1085/jgp.93.5.979
979
999
Journal of General Physiology
93
Ionic currents of enzymatically dispersed type 1 and type 11 cells of the carotid body have been studied using the whole cell variant of the patch-clamp technique. Type 11 cells only have a tiny, slowly activating outward potassium cur rent. By contrast, in every type 1 chemoreceptor cell studied we found (a) sodium,
(b) calcium, and (e) potassium currents. (a) The sodium current has a fast activation time course and an activation threshold at --40 mV. At ali voltages inactivation follows a single exponential time course. The time constant of inactivation is 0.67 ms at O mV. Half steady state inactivation occurs at a membrane potential of
--50 mV. (b) The calcium current is almost totally abolished when most of the extemal calcium is replaced by magnesium. The activation threshold of this cur rent is at --40 mV and at O mV it reaches a peak amplitude in 6-8 ms. The calcium current inactivates very slowly and only decreases to 27% of the maximal value at the end of 300-ms pulses to 40 mV. The calcium current was about two times larger when barium ions were used as charge carriers instead of calcium ions. Barium ions also shifted 15-20 mV toward negative voltages the conductance vs. voltage curve. Deactivation kinetics of the calcium current follows a biphasic time course well fitted by the sum of two exponentials. At -80 mV the slow com ponent has a time constant of 1.3 ± 0.4 ms whereas the fast component, with an amplitude about 20 times larger than the slow component, has a time constant of
0.16 ± 0.03 ms. These results suggest that type 1 cells have predominantly fast deactivating calcium channels. The slow component of the tails may represent the activity of a small population of slowly deactivating calcium channels, although other possibilities are considered. (e) Potassium current seems to be mainly due to the activity of voltage-dependent potassium channels, but a small percentage of calcium-activated channels may also exist. This current activates slowly, reaches a peak amplitude in 5-1O ms, and thereafter slowly inactivates. Inactivation is almost complete in 250-300 ms. The potassium current is reversibly blocked by tetraeth ylammonium. Under current-clamp conditions type I cells can spontaneously fire large action potentials.
eng
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http://creativecommons.org/licenses/by-nc-nd/4.0/
© 1989 The Rockefeller University Press
Attribution-NonCommercial-NoDerivatives 4.0 International
Ionic currents in dispersed chemoreceptor cells of the mammalian carotid body
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/250162021-05-21T21:03:53Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
López López, José Ramón
Shacklock, Philip S.
Balke, C. William
Gil, Withrowl
2017-08-23T11:37:27Z
2017-08-23T11:37:27Z
1995
Science, vol.268, n.5213 p.1042-1045
0036-8075
http://uvadoc.uva.es/handle/10324/25016
1042
1045
Science
268
Excitation-contraction coupling was studied in mammalian cardiac cells in which the opening probability of L-type calcium (Ca2 +) channels was reduced. Confocal microscopy during voltage-clamp depolarization revealed distinct local transients inthe concentration of intracellular calcium ions ([Ca2 +]¡). When voltage was varied, the latency to occurrence and the relative probability of occurrence of local [Ca2 +]; transients varied as predicted if ca2 + release from the sarcoplasmic reticulum (SR) was linked tightly to Ca2 + flux through L-type Ca2 + channels but not to that through the Na-Ca exchanger or to average [Ca2 +];. Voltage had no effect on the amplitude of local [Ca2 +]; transients. Thus, the most efficacious "Ca2 + signal" for activating Ca2 + release from the SR may be a transient microdomain of high [Ca2 +]; beneath an individual, open L-type Ca2 + channel
eng
info:eu-repo/semantics/openAccess
Local Calcium Transients Triggered by Single L-Type Calcium Channel Currents in Cardiac Cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/253272021-10-18T12:37:59Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
Tamayo Lomas, Luis Mariano
López López, José Ramón
Castañeda, Javier
2017-08-31T10:05:19Z
2017-08-31T10:05:19Z
1997
Pflügers Arch - Eur J. Physiol (1997)
0031-6768
http://uvadoc.uva.es/handle/10324/25327
698
704
Pflügers Arch - Eur J. Physiol
434
Hypoxia activates erythropoietin-producing cells, chemoreceptor cells of the carotid body and pul- monary artery smooth muscle cells (PSMC) with a com-
Key words Hypoxia · Pulmonary artery · Carbon monoxide&bdy:
parable arterial PO2 threshold of some 70 mmHg. The
inhibition by CO of the hypoxic responses in the two for- mer cell types has led to the proposal that a haemopro- tein is involved in the detection of the PO2 levels. Here, we report the effect of CO on the hypoxic pulmonary va- soconstriction (HPV). Pulmonary arterial pressure (PAP)
was measured in an in situ, blood-perfused lung prepara- tion. PAP in normoxia (20% O2, 5% CO2) .
eng
info:eu-repo/semantics/openAccess
Monóxido de carbono
Hypoxia
Carbon Monoxide inhibits hypoxic pulmonary vasoconstriction in rats by a cGMP-independent mechanism
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/259192021-06-24T07:36:26Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
Pérez García, María Teresa
López López, José Ramón
González, Constancio
2017-09-26T08:05:55Z
2017-09-26T08:05:55Z
1999
Journal of Genetic Physiology,1999, vol. 113. p. 897-907
0022-1295
http://uvadoc.uva.es/handle/10324/25919
10.1085/jgp.113.6.897
897
907
Journal of General Physiology
113
Voltage-gated K+ (KV) channels are protein complexes composed of ion-conducting integral membrane α subunits and cytoplasmic modulatory β subunits. The differential expression and association of α and β subunits seems to contribute significantly to the complexity and heterogeneity of KV channels in excitable cells, and their functional expression in heterologous systems provides a tool to study their regulation at a molecular level. Here, we have studied the effects of Kvβ1.2 coexpression on the properties of Shaker and Kv4.2 KV channel α subunits, which encode rapidly inactivating A-type K+ currents, in transfected HEK293 cells. We found that Kvβ1.2 functionally associates with these two α subunits, as well as with the endogenous KV channels of HEK293 cells, to modulate different properties of the heteromultimers. Kvβ1.2 accelerates the rate of inactivation of the Shaker currents, as previously described, increases significantly the amplitude of the endogenous currents, and confers sensitivity to redox modulation and hypoxia to Kv4.2 channels. Upon association with Kvβ1.2, Kv4.2 can be modified by DTT (1,4 dithiothreitol) and DTDP (2,2′-dithiodipyridine), which also modulate the low pO2 response of the Kv4.2+β channels. However, the physiological reducing agent GSH (reduced glutathione) did not mimic the effects of DTT. Finally, hypoxic inhibition of Kv4.2+β currents can be reverted by 70% in the presence of carbon monoxide and remains in cell-free patches, suggesting the presence of a hemoproteic O2 sensor in HEK293 cells and a membrane-delimited mechanism at the origin of hypoxic responses. We conclude that β subunits can modulate different properties upon association with different KV channel subfamilies; of potential relevance to understanding the molecular basis of low pO2 sensitivity in native tissues is the here described acquisition of the ability of Kv4.2+β channels to respond to hypoxia.
eng
info:eu-repo/semantics/openAccess
© 1999 Rockefeller University Press
Kvβ1.2 subunit coexpression in HEK293 cells confers O2 sensitivity to Kv4.2 but not to shaker channels
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/259502021-06-24T07:36:39Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
Pérez García, María Teresa
López López, José Ramón
Riesco Fagundo, Armenia María
Hoppe, Uta C.
Marbán, Eduardo
González, Constancio
Johns, David C.
2017-09-27T10:01:28Z
2018-03-27T23:40:31Z
2000
Journal of Neuroscience, 2000, vol. 20, n. 15. p. 5689-5695
5689-5695
http://uvadoc.uva.es/handle/10324/25950
10.1523/JNEUROSCI.20-15-05689.2000
5689
15
5695
2o
Hypoxia initiates the neurosecretory response of the carotid body (CB) by inhibiting one or more potassium channels in the chemoreceptor cells. Oxygen-sensitive K+ channels were first described in rabbit CB chemoreceptor cells, in which a transient outward K+ current was reported to be reversibly inhibited by hypoxia. Although progress has been made to characterize this current with electrophysiological and pharmacological tools, no attempts have been made to identify which Kv channel proteins are expressed in rabbit CB chemoreceptor cells and to determine
+
with adenoviruses that enabled ecdysone-inducible expression of the dominant-negative constructs and reporter genes in poly- cistronic vectors. In voltage-clamp experiments, we found that, whereas adenoviral infections of chemoreceptor cells with Kv1.xDN did not modify the O -sensitive K+ current, infections with Kv4.xDN suppressed the transient outward current in a time-dependent manner, significantly depolarized the cells, and abolished the depolarization induced by hypoxia. Our work dem- onstrate that genes of the Shal K+ channels underlie the tran-
+
their contribution to the native O2-sensitive K
current. To probe
sient outward, O2-sensitive, K
current of rabbit CB chemore-
the molecular identity of this current, we have used dominant-
negative constructs to block the expression of functional Kv channels of the Shaker (Kv1.xDN) or the Shal (Kv4.xDN) subfam-
ceptor cells and that this current contributes to the cell
depolarization in response to low pO2.
+
ilies, because members of these two subfamilies contribute to
Key words: O2-sensitive K
current; viral gene transfer;
the transient outward K+ currents in other preparations. Delivery of the constructs into chemoreceptor cells has been achieved
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2000 The Society for Neuroscience
Attribution-NonCommercial-NoDerivatives 4.0 International
Viral gene transfer of dominant-negative Kv4 construct suppresses an O2-sensitive K+ current in chemoreceptor cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/291732021-06-24T07:36:24Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
López López, José Ramón
Colinas, Olaia
Pérez Carretero, Francisco D.
Pérez García, María Teresa
2018-03-22T12:07:06Z
2018-03-22T12:07:06Z
2008
The Journal of General Physiology, April 2008, vol. 131, n. 5. p. 455-471
0022-1295
http://uvadoc.uva.es/handle/10324/29173
https://doi.org/10.1085/jgp.200709912
455
5
471
The Journal of General Physiology
131
Shal-type (Kv4) channels are expressed in a large variety of tissues, where they contribute to transient voltage- dependent K+ currents. Kv4 are the molecular correlate of the A-type current of neurons (ISA), the fast component of ITO current in the heart, and also of the oxygen-sensitive K+ current (KO2) in rabbit carotid body (CB) chemore- ceptor cells. The enormous degree of variability in the physiological properties of Kv4-mediated currents can be attributable to the complexity of their regulation together with the large number of ancillary subunits and scaffold- ing proteins that associate with Kv4 proteins to modify their trafficking and their kinetic properties. Among those, KChIPs and DPPX proteins have been demonstrated to be integral components of ISA and ITO currents, as their co- expression with Kv4 subunits recapitulates the kinetics of native currents. Here, we explore the presence and func- tional contribution of DPPX to KO2 currents in rabbit CB chemoreceptor cells by using DPPX functional knockdown with siRNA. Additionally, we investigate if the presence of DPPX endows Kv4 channels with new pharmacological properties, as we have observed anomalous tetraethylammonium (TEA) sensitivity in the native KO2 currents. DPPX association with Kv4 channels induced an increased TEA sensitivity both in heterologous expression systems and in CB chemoreceptor cells. Moreover, TEA application to Kv4-DPPX heteromultimers leads to marked kinetic ef- fects that could be explained by an augmented closed-state inactivation. Our data suggest that DPPX proteins are integral components of KO2 currents, and that their association with Kv4 subunits modulate the pharmacological profile of the heteromultimers.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Neurofisiología
A Role for DPPX Modulating External TEA Sensitivity of Kv4 Channels
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/291872021-06-24T07:36:36Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
López López, José Ramón
Alpizar, Yeranddy A.
Meseguer, Víctor
Enoch, Luis
Tajada Esteban, Sendoa
Denlinger, Bristol
Fajardo, Otto
Manenschijn, Jan-Albert
Fernández Peña, Carlos
Talavera, Arturo
Kichko, Tatiana
Navia, Belén
Sánchez, Alicia
Señarís, Rosa
Reeh, Peter
Pérez García, María Teresa
Voets, Thomas
Belmonte, Carlos
Talavera, Karel
Viana, Felix
2018-03-23T10:21:30Z
2018-03-23T10:21:30Z
2014
Nature Communications, 2014, vol. 5. p. 1-16
2041-1723
http://uvadoc.uva.es/handle/10324/29187
10.1038/ncomms4125
1
16
Nature Communications
5
Gram-negative bacterial infections are accompanied by inflammation and somatic or visceral pain. These symptoms are generally attributed to sensitization of nociceptors by inflammatory mediators released by immune cells. Nociceptor sensitization during inflammation occurs through activation of the Toll-like receptor 4 (TLR4) signalling pathway by lipopolysaccharide (LPS), a toxic by-product of bacterial lysis. Here we show that LPS exerts fast, membrane delimited, excitatory actions via TRPA1, a transient receptor potential cation channel that is critical for transducing environmental irritant stimuli into nociceptor activity. Moreover, we find that pain and acute vascular reactions, including neurogenic inflammation (CGRP release) caused by LPS are primarily dependent on TRPA1 channel activation in nociceptive sensory neurons, and develop independently of TLR4 activation. The identification of TRPA1 as a molecular determinant of direct LPS effects on nociceptors offers new insights into the pathogenesis of pain and neurovascular responses during bacterial infections and opens novel avenues for their treatment.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
TRPA1 channels mediate acute neurogenic inflammation and pain produced by bacterial endotoxins
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/291912021-06-24T07:36:38Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
López López, José Ramón
Fernández Mariño, Ana Isabel
Cidad Velasco, María del Pilar
Zafra, Delia
Nocito, Laura
Domínguez, Jorge
Oliván Viguera, Aida
Köhler, Ralf
Pérez García, María Teresa
Valverde, Miguel Ángel
Guinovart, Joan J.
Fernández Fernández, José Manuel
2018-03-23T12:45:02Z
2018-03-23T12:45:02Z
2015
Plos One, 2015, p. 1-21
1932-6203
http://uvadoc.uva.es/handle/10324/29191
https://doi.org/10.1371/journal.pone.0118148
1
21
Despite the substantial knowledge on the antidiabetic, antiobesity and antihypertensive actions of tungstate, information on its primary target/s is scarce. Tungstate activates both the ERK1/2 pathway and the vascular voltage- and Ca2+-dependent large-conductance BKαβ1 potassium channel, which modulates vascular smooth muscle cell (VSMC) proliferation and function, respectively. Here, we have assessed the possible involvement of BKαβ1 channels in the tungstate-induced ERK phosphorylation and its relevance for VSMC proliferation. Western blot analysis in HEK cell lines showed that expression of vascular BKαβ1 channels potentiates the tungstate-induced ERK1/2 phosphorylation in a Gi/o protein-dependent manner. Tungstate activated BKαβ1 channels upstream of G proteins as channel activation was not altered by the inhibition of G proteins with GDPβS or pertussis toxin. Moreover, analysis of Gi/o protein activation measuring the FRET among heterologously expressed Gi protein subunits suggested that tungstate-targeting of BKαβ1 channels promotes G protein activation. Single channel recordings on VSMCs from wild-type and β1-knockout mice indicated that the presence of the regulatory β1 subunit was essential for the tungstate-mediated activation of BK channels in VSMCs. Moreover, the specific BK channel blocker iberiotoxin lowered tungstate-induced ERK phosphorylation by 55% and partially reverted (by 51%) the tungstate-produced reduction of platelet-derived growth factor (PDGF)-induced proliferation in human VSMCs. Our observations indicate that tungstate-targeting of BKαβ1 channels promotes activation of PTX-sensitive Gi proteins to enhance the tungstate-induced phosphorylation of ERK, and inhibits PDGF-stimulated cell proliferation in human vascular smooth muscle.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Tungstate-Targeting of BKαβ1 Channels Tunes ERK Phosphorylation and Cell Proliferation in Human Vascular Smooth Muscle
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/292092022-06-28T11:42:44Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
Jiménez Pérez, Laura
Cidad Velasco, María del Pilar
Álvarez Miguel, Inés
Santos Hipólito, Alba
Torres Merino, Rebeca
Alonso Alonso, Esperanza
Fuente García, Miguel Ángel de la
Pérez García, María Teresa
López López, José Ramón
2018-03-26T10:31:02Z
2018-03-26T10:31:02Z
2016
The journal of biological chemistry, Febrero 2016, vol. 291, n. 7, p. 3569-3580
0021-9258
http://uvadoc.uva.es/handle/10324/29209
10.1074/jbc.M115.678995
3569
7
3580
The journal of biological chemistry
291
Changes in voltage-dependent potassium channels (Kv channels) associate to proliferation in many cell types, including transfected HEK293 cells. In this system Kv1.5 overexpression decreases proliferation, whereas Kv1.3 expression increases it independently of K+ fluxes. To identify Kv1.3 domains involved in a proliferation-associated signaling mechanism(s), we constructed chimeric Kv1.3-Kv1.5 channels and point-mutant Kv1.3 channels, which were expressed as GFP- or cherry-fusion proteins. We studied their trafficking and functional expression, combining immunocytochemical and electrophysiological methods, and their impact on cell proliferation. We found that the C terminus is necessary for Kv1.3-induced proliferation. We distinguished two residues (Tyr-447 and Ser-459) whose mutation to alanine abolished proliferation. The insertion into Kv1.5 of a sequence comprising these two residues increased proliferation rate. Moreover, Kv1.3 voltage-dependent transitions from closed to open conformation induced MEK-ERK1/2-dependent Tyr-447 phosphorylation. We conclude that the mechanisms for Kv1.3-induced proliferation involve the accessibility of key docking sites at the C terminus. For one of these sites (Tyr-447) we demonstrated the contribution of MEK/ERK-dependent phosphorylation, which is regulated by voltage-induced conformational changes.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Molecular Determinants of Kv1.3 Potassium Channels-induced Proliferation
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/390052021-06-24T07:36:34Zcom_10324_32522com_10324_952com_10324_894com_10324_43677com_10324_954com_10324_1134com_10324_931col_10324_32523col_10324_43678col_10324_1213
Arévalo Martínez, Marycarmen
Cidad Velasco, María del Pilar
García Mateo, Nadia
Moreno Estar, Sara
Serna Pérez, Julia
Fernández, Mirella
Swärd, Karl
Simarro Grande, María
Fuente García, Miguel Ángel de la
López López, José Ramón
Pérez García, María Teresa
2019-11-04T09:36:19Z
2019-11-04T09:36:19Z
2019
Arterioscler Thromb Vasc Biol. 2019 Oct 10:ATVBAHA119313492. doi: 10.1161/ATVBAHA.119.313492
1079-5642
http://uvadoc.uva.es/handle/10324/39005
10.1161/ATVBAHA.119.313492
Arteriosclerosis, Thrombosis, and Vascular Biology
1524-4636
Objective: We have previously described that changes in the expression of Kv channels associate to phenotypic modulation (PM), so that Kv1.3 /Kv1.5 ratio is a landmark of vascular smooth muscle cells (VSMCs) phenotype. Moreover, we demonstrated that the Kv1.3 functional expression is relevant for PM in several types of vascular lesions. Here, we explore the efficacy of Kv1.3 inhibition for the prevention of remodeling in human vessels, and the mechanisms linking the switch in Kv1.3 /Kv1.5 ratio to PM.
Approach and Results: Vascular remodeling was explored using organ culture and primary cultures of VSMCs obtained from human vessels. We studied the effects of Kv1.3 inhibition on serum-induced remodeling, as well as the impact of viral vector-mediated overexpression of Kv channels or myocardin knock-down. Kv1.3 blockade prevented remodeling by inhibiting proliferation, migration and extracellular matrix (ECM) secretion. PM activated Kv1.3 via downregulation of Kv1.5. Hence, both Kv1.3 blockers and Kv1.5 overexpression inhibited remodeling in a non-additive fashion. Finally, myocardin knock-down induced vessel remodeling and Kv1.5 downregulation and myocardin overexpression increased Kv1.5, while Kv1.5 overexpression inhibited PM without changing myocardin expression.
Conclusions: We demonstrate that Kv1.5 channel gene is a myocardin-regulated, VSMCs contractile marker. Kv1.5 downregulation upon PM leaves Kv1.3 as the dominant Kv1 channel expressed in dedifferentiated cells. We demonstrated that the inhibition of Kv1.3 channel function with selective blockers or by preventing Kv1.5 downregulation can represent an effective, novel strategy for the prevention of intimal hyperplasia and restenosis of the human vessels used for coronary angioplasty procedures.
spa
info:eu-repo/semantics/openAccess
Myocardin-Dependent Kv1.5 Channel Expression Prevents Phenotypic Modulation of Human Vessels in Organ Culture
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/409992021-06-23T09:52:36Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Muñoz Martínez, Raquel
Hileeto, Denise
Cruz Muñoz, William
Wood, Geoffrey A.
Xu, Ping
Man, Shan
Viloria Petit, Alicia
Kerbel, Robert S.
2020-06-05T12:27:36Z
2020-06-05T12:27:36Z
2019
PLoS ONE, 2019, vol. 14, n. 9. 28 p.
1932-6203
http://uvadoc.uva.es/handle/10324/40999
10.1371/journal.pone.0222580
Metronomic chemotherapy using the 5-FU prodrug uracil-tegafur (UFT) and cyclophosphamide (CTX) was previously shown to only modestly delay primary tumor growth, but nevertheless markedly suppressed the development of micro-metastasis in an orthotopic breast cancer xenograft model, using the metastatic variant of the MDA-MB-231 cell line, 231/LM2-4. Furthermore, a remarkable prolongation of survival, with no toxicity, was observed in a model of postsurgical advanced metastatic disease. A question that has remained unanswered is the seemingly selective anti-metastatic mechanisms of action responsible for this treatment. We assessed the in vivo effect of metronomic UFT, CTX or their combination, on vascular density, collagen deposition and c-Met (cell mediators or modulators of tumor cell invasion or dissemination) via histochemistry/immunohistochemistry of primary tumor sections. We also assessed the effect of continuous exposure to low and non-toxic doses of active drug metabolites 5-fluorouracil (5-FU), 4-hydroperoxycyclophosphamide (4-HC) or their combination, on 231/LM2-4 cell invasiveness in vitro. In the in vivo studies, a significant reduction in vascular density and p-Met[Y1003] levels was associated with UFT+CTX treatment. All treatments reduced intratumoral collagen deposition. In the in vitro studies, a significant reduction of collagen IV invasion by all treatments was observed. The 3D structures formed by 231/LM2-4 on Matrigel showed a predominantly Mass phenotype under treated conditions and Stellate phenotype in untreated cultures. Taken together, the results suggest the low-dose metronomic chemotherapy regimens tested can suppress several mediators of tumor invasiveness highlighting a new perspective for the anti-metastatic efficacy of metronomic chemotherapy.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2019 PLOS
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Suppressive impact of metronomic chemotherapy using UFT and/or cyclophosphamide on mediators of breast cancer dissemination and invasion
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/410002021-06-23T09:52:38Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Muñoz Martínez, Raquel
Man, Shan
Shaked, Yuval
Lee, Christina R.
Wong, John
Francia, Giulio
Kerbel, Robert S.
2020-06-05T12:50:37Z
2020-06-05T12:50:37Z
2006
Cancer Research, 2006, vol. 66, n. 7. p. 3386-3391
1538-7445
http://uvadoc.uva.es/handle/10324/41000
10.1158/0008-5472.CAN-05-4411
Metronomic antiangiogenic chemotherapy, the prolonged
administration of relatively low drug doses, at close regular
intervals with no significant breaks, has been mainly studied
at the preclinical level using single chemotherapeutic drugs,
frequently in combination with a targeted antiangiogenic
drug, and almost always evaluated on primary localized
tumors. We tested a ‘‘doublet’’ combination metronomic
chemotherapy treatment using two oral drugs, UFT, a
5-fluorouracil (5-FU) prodrug administered by gavage, and
cyclophosphamide, for efficacy and toxicity in a new mouse
model of advanced, terminal, metastatic human breast cancer.
The optimal biological dose of each drug was first determined
by effects on levels of circulating endothelial progenitor cells
as a surrogate marker for angiogenesis, which was assessed to
be 15 mg/kg for UFT and 20 mg/kg for cyclophosphamide. A
combination treatment was then evaluated in mice with
advanced metastatic disease using a serially selected metastatic variant of the MDA-MB-231 breast cancer-cell line, 231/
LM2-4. UFT or cyclophosphamide treatment showed only very
modest survival advantages whereas a combination of the two
resulted in a remarkable prolongation of survival, with no
evidence of overt toxicity despite 140 days of continuous
therapy, such that a significant proportion of mice survived
for over a year. In contrast, this striking therapeutic effect of
the combination treatment was not observed when tested on
primary orthotopic tumors. We conclude that combination
oral low-dose daily metronomic chemotherapy, using cyclophosphamide and UFT, is superior to monotherapy and seems
to be a safe and highly effective experimental antimetastatic
therapy, in this case, for advanced metastatic breast cancer
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2006 American Association for Cancer Research
Attribution-NonCommercial-NoDerivs 3.0 Unported
Highly efficacious nontoxic preclinical treatment for advanced metastatic breast cancer using combination oral UFT-cyclophosphamide metronomic chemotherapy
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/410042021-06-23T09:52:40Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Klingenberg, Olav
Wiedlocha, Antoni
Rapak, Andrzej
Muñoz Martínez, Raquel
Falnes, Pål
Olsnes, Sjur
2020-06-08T12:13:19Z
2020-06-08T12:13:19Z
1998
Journal of Biological Chemistry, 1998, vol. 273, n. 18. p. 11164-11172
0021-9258
http://uvadoc.uva.es/handle/10324/41004
10.1074/jbc.273.18.11164
Acidic fibroblast growth factor (aFGF) is a potent mitogen. It acts through activation of specific cell surface receptors leading to intracellular tyrosine phosphorylation cascades, but several reports also indicate that aFGF enters cells and that it has an intracellular function as well. The aFGF(K132E) mutant binds to and activates fibroblast growth factor receptors equally strongly as the wild-type, but it is a poor mitogen. We demonstrate that aFGF(K132E) enters NIH 3T3 cells and is transported to the nuclear fraction like wild-type aFGF. A fusion protein of aFGF(K132E) and diphtheria toxin A-fragment (aFGF(K132E)-DT-A) and a similar fusion protein containing wild-type aFGF (aFGF-DT-A) were reconstituted with diphtheria toxin B-fragment. Both fusion proteins were translocated to the cytosol by the diphtheria toxin pathway and subsequently recovered from the nuclear fraction. Whereas translocation of aFGF-DT-A stimulated DNA synthesis in U2OSDR1 cells lacking functional fibroblast growth factor receptors, aFGF(K132E)-DT-A did not. The mutation disrupts a protein kinase C phosphorylation site in the growth factor making it unable to be phosphorylated. The data indicate that a defect in the intracellular action of aFGF(K132E) is the reason for its strongly reduced mitogenicity, possibly due to inability to be phosphorylated.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 1998 American Society for Biochemistry and Molecular Biology
Attribution-NonCommercial-NoDerivs 3.0 Unported
Inability of the acidic fibroblast growth factor mutant K132E to stimulate DNA synthesis after translocation into cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/410342021-06-23T09:52:42Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Muñoz Martínez, Raquel
Klingenberg, Olav
Wiedlocha, Antoni
Rapak, Andrzej
Falnes, Pål
Olsnes, Sjur
2020-06-12T11:20:51Z
2020-06-12T11:20:51Z
1997
Oncogene, 1997, vol. 15, n. 5. p. 525-536
1476-5594
http://uvadoc.uva.es/handle/10324/41034
10.1038/sj.onc.1201226
Acidic fibroblast growth factor (aFGF) binds to specific transmembrane receptors and is partly transported to a nuclear location. To study this transport we made a kinase-negative mutant of FGF receptor 4 as well as one where the major part of the cytoplasmic receptor domain was deleted, and expressed them in U2OSDr1 cells that lack functional FGF receptors. All receptors mediated endocytic uptake of aFGF. Translocation of the growth factor across cellular membranes was assayed using aFGF with a C-terminal CAAX-motif, which signals addition of a farnesyl group onto the protein once in the cytosol. CAAX-tagged aFGF was farnesylated when incubated with cells containing wild-type or kinase-negative receptors. It was not farnesylated in cells expressing the deleted receptor, or when the incubation was in the presence of genistein. aFGF incubated with cells transfected with wild-type or kinase-negative receptors, but not with the deleted receptor, was partly recovered from the nuclear fraction in the absence, but not in the presence of genistein. The data indicate that the cytoplasmic receptor domain, but not the active kinase, is required for transport of the growth factor into cells, and that genistein inhibits the process.
eng
info:eu-repo/semantics/openAccess
© 1997 Springer Nature
Effect of mutation of cytoplasmic receptor domain and of genistein on transport of acidic fibroblast growth factor into cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/410052021-06-23T09:52:41Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Wiedlocha, Antoni
Falnes, Pål
Rapak, Andrzej
Muñoz Martínez, Raquel
Klingenberg, Olav
Olsnes, Sjur
2020-06-08T13:15:28Z
2020-06-08T13:15:28Z
1996
Molecular and Cellular Biology, 1996, vol. 16, n. 1. p. 270-280
0270-7306
http://uvadoc.uva.es/handle/10324/41005
10.1128/MCB.16.1.270
U2OS Dr1 cells, originating from a human osteosarcoma, are resistant to the intracellular action of diphtheria toxin but contain toxin receptors on their surfaces. These cells do not have detectable amounts of fibroblast growth factor receptors. When these cells were transfected with fibroblast growth factor receptor 4, the addition of acidic fibroblast growth factor to the medium induced tyrosine phosphorylation, DNA synthesis, and cell proliferation. A considerable fraction of the cell-associated growth factor was found in the nuclear fraction. When the growth factor was fused to the diphtheria toxin A fragment, it was still bound to the growth factor receptor and induced tyrosine phosphorylation but did not induce DNA synthesis or cell proliferation, nor was any fusion protein recovered in the nuclear fraction. On the other hand, when the fusion protein was associated with the diphtheria toxin B fragment to allow translocation to the cytosol by the toxin pathway, the fusion protein was targeted to the nucleus and stimulated both DNA synthesis and cell proliferation. In untransfected cells containing toxin receptors but not fibroblast growth factor receptors, the fusion protein was translocated to the cytosol and targeted to the nucleus, but in this case, it stimulated only DNA synthesis. These data indicate that the following two signals are required to stimulate cell proliferation in transfected U2OS Dr1 cells: the tyrosine kinase signal from the activated fibroblast growth factor receptor and translocation of the growth factor into the cell.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 1996 American Society for Microbiology
Attribution-NonCommercial-NoDerivs 3.0 Unported
Stimulation of proliferation of a human osteosarcoma cell line by exogenous acidic fibroblast growth factor requires both activation of receptor tyrosine kinase and growth factor internalization
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425832021-06-23T09:52:44Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Yin, Yi
Sedlaczek, Oliver
Muller, Benedikt
Warth, Arne
González-Vallinas Garrachón, Margarita
Lahrmann, Bernd
Grabe, Niels
Kauczor, Hans-Ulrich
Breuhahn, Kai
Vignon Clementel, Irene E.
Drasdo, Dirk
2020-09-28T05:41:43Z
2020-09-28T05:41:43Z
2018
IEEE Transactions on Medical Imaging, 2018, vol. 37, n. 1 p. 35-46
0278-0062
http://uvadoc.uva.es/handle/10324/42583
10.1109/TMI.2017.2698525
35
1
46
IEEE Transactions on Medical Imaging
37
1558-254X
Diffusion-weighted magnetic resonance imaging (DWI) is a key non-invasive imaging technique for cancer diagnosis and tumor treatment assessment, reflecting Brownian movement of water molecules in tissues. Since densely packed cells restrict molecule mobility, tumor tissues produce usually higher signal (a.k.a. less attenuated signal) on isotropic maps compared with normal tissues. However, no general quantitative relation between DWI data and the cell density has been established. In order to link low-resolution clinical cross-sectional data with high-resolution histological information, we developed an image processing and analysis chain, which was used to study the correlation between the diffusion coefficient (D value) estimated from DWI and tumor cellularity from serial histological slides of a resected non-small cell lung cancer tumor. Color deconvolution followed by cell nuclei segmentation was performed on digitized histological images to determine local and cell-type specific 2d (two-dimensional) densities. From these, the 3d cell density was inferred by a model-based sampling technique, which is necessary for the calculation of local and global 3d tumor cell count. Next, DWI sequence information was overlaid with high-resolution CT data and the resected histology using prominent anatomical hallmarks for co-registration of histology tissue blocks and non-invasive imaging modalities' data. The integration of cell numbers information and DWI data derived from different tumor areas revealed a clear negative correlation between cell density and D value. Importantly, spatial tumor cell density can be calculated based on DWI data. In summary, our results demonstrate that tumor cell count and heterogeneity can be predicted from DWI data, which may open new opportunities for personalized diagnosis and therapy optimization.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© IEEE
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Tumor cell load and heterogeneity estimation from diffusion-weighted MRI calibrated with histological data: an example from lung cancer
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425842021-06-23T09:52:46Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Lotz, J. M.
Olesch, J.
Muller, Benedikt
Polzin, T.
Galuschka, P.
Lotz, J. M.
Heldmann, S
Laue, H
González-Vallinas Garrachón, Margarita
Warth, Arne
Lahrmann, Bernd
Grabe, N
Sedlaczek, Oliver
Breuhahn, Kai
Modersitzki, J
2020-09-28T05:46:07Z
2020-09-28T05:46:07Z
2016
IEEE Transactions on biomedical engineering, 2016, Vol. 63, n. 9, p.1812-1819
0018-9294
http://uvadoc.uva.es/handle/10324/42584
10.1109/TBME.2015.2503122
1812
9
1819
IEEE Transactions on Biomedical Engineering
63
1558-2531
Image registration of whole slide histology images allows the fusion of fine-grained information-like different immunohistochemical stains-from neighboring tissue slides. Traditionally, pathologists fuse this information by looking subsequently at one slide at a time. If the slides are digitized and accurately aligned at cell level, automatic analysis can be used to ease the pathologist's work. However, the size of those images exceeds the memory capacity of regular computers. Methods: We address the challenge to combine a global motion model that takes the physical cutting process of the tissue into account with image data that is not simultaneously globally available. Typical approaches either reduce the amount of data to be processed or partition the data into smaller chunks to be processed separately. Our novel method first registers the complete images on a low resolution with a nonlinear deformation model and later refines this result on patches by using a second nonlinear registration on each patch. Finally, the deformations computed on all patches are combined by interpolation to form one globally smooth nonlinear deformation. The NGF distance measure is used to handle multistain images. Results: The method is applied to ten whole slide image pairs of human lung cancer data. The alignment of 85 corresponding structures is measured by comparing manual segmentations from neighboring slides. Their offset improves significantly, by at least 15%, compared to the low-resolution nonlinear registration. Conclusion/Significance: The proposed method significantly improves the accuracy of multistain registration which allows us to compare different antibodies at cell level.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© IEEE
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Patch-based nonlinear image registration for gigapixel whole slide images
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425852021-06-23T09:52:47Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González-Vallinas Garrachón, Margarita
Breuhahn, Kai
2020-09-28T06:49:40Z
2020-09-28T06:49:40Z
2016
HepatoBiliary Surgery and Nutrition, 2016, Vol. 5, n. 4, p. 372-376
2304-3881
http://uvadoc.uva.es/handle/10324/42585
10.21037/hbsn.2016.05.07
372
4
376
HepatoBiliary Surgery and Nutrition
5
2304-389X
Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death worldwide, and it is well accepted that the poor outcome of HCC patients among others is caused by metastasis and tumor cell dissemination.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© AME Publishing Company
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
MicroRNAs are key regulators of hepatocellular carcinoma (HCC) cell dissemination—what we learned from microRNA-494
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425862021-06-23T09:52:49Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González-Vallinas Garrachón, Margarita
Rodríguez Paredes, Manuel
Albrecht, Marco
Sticht, Carsten
Stichel, Damian
Gutekunst, Julian
Pitea, Adriana
Sass, Steffen
Sánchez Rivera, Francisco J.
Lorenzo Bermejo, Justo
Schmitt, Jennifer
Torre, Carolina de la
Warth, Arne
Theis, Fabian J.
Müller, Nikola S.
Gretz, Norbert
Muley, Thomas
Meister, Michael
Tschaharganeh, Darjus F.
Schirmacher, Peter
Matthäus, Franziska
Breuhahn, Kai
2020-09-28T06:52:45Z
2020-09-28T06:52:45Z
2018
Molecular Cancer Research, 2018, vol. 16, n. 3, p. 390-402
1541-7786
http://uvadoc.uva.es/handle/10324/42586
10.1158/1541-7786.MCR-17-0334
390
3
402
Molecular Cancer Research
16
1557-3125
Most lung cancer deaths are related to metastases, which indicates the necessity of detecting and inhibiting tumor cell dissemination. Here, we aimed to identify miRNAs involved in metastasis of lung adenocarcinoma as prognostic biomarkers and therapeutic targets. To that end, lymph node metastasis–
associated miRNAs were identified in The Cancer Genome Atlas lung adenocarcinoma patient cohort (sequencing data; n ¼ 449) and subsequently validated by qRT-PCR in an independent
clinical cohort (n ¼ 108). Overexpression of miRNAs located
on chromosome 14q32 was associated with metastasis in lung
adenocarcinoma patients. Importantly, Kaplan–Meier analysis
and log-rank test revealed that higher expression levels of
individual 14q32 miRNAs (mir-539, mir-323b, and mir487a) associated with worse disease-free survival of never-smoker patients. Epigenetic analysis including DNA methylation
microarray data and bisulfite sequencing validation demonstrated that the induction of 14q32 cluster correlated with genomic
hypomethylation of the 14q32 locus. CRISPR activation technology, applied for the first time to functionally study the
increase of clustered miRNA levels in a coordinated manner,
showed that simultaneous overexpression of 14q32 miRNAs
promoted tumor cell migratory and invasive properties. Analysis
of individual miRNAs by mimic transfection further illustrated
that miR-323b-3p, miR-487a-3p, and miR-539-5p significantly
contributed to the invasive phenotype through the indirect
regulation of different target genes. In conclusion, overexpression of 14q32 miRNAs, associated with the respective genomic
hypomethylation, promotes metastasis and correlates with poor
patient prognosis in lung adenocarcinoma
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© American Association for Cancer Research
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Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Epigenetically regulated chromosome 14q32 miRNA cluster induces metastasis and predicts poor prognosis in lung adenocarcinoma patients
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425872021-06-23T09:52:50Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ramírez de Molina, Ana
Vargas, Teodoro
Molina, Susana
Sánchez, Jenifer
Martínez Romero, Jorge
González-Vallinas Garrachón, Margarita
Martín Hernández, Roberto
Sánchez Martínez, Ruth
Gómez de Cedrón, Marta
Dávalos, Alberto
Calani, Luna
Rio, Daniele del
González Sarrías, Antonio
Espín, Juan Carlos
Tomás Barberán, Francisco A.
Reglero, Guillermo
2020-09-28T07:02:04Z
2020-09-28T07:02:04Z
2015
Journal of Pharmacology and Experimental Therapeutics, 2015, vol. 353, n. 2, p. 433-444
0022-3565
http://uvadoc.uva.es/handle/10324/42587
10.1124/jpet.114.221796
433
2
444
Journal of Pharmacology and Experimental Therapeutics
353
1521-0103
Ellagic acid (EA) and some derivatives have been reported to inhibit cancer cell proliferation, induce cell cycle arrest, and modulate some important cellular processes related to cancer. This study aimed to identify possible structure-activity relationships of EA and some in vivo derivatives in their antiproliferative effect on both human colon cancer and normal cells, and to compare this activity with that of other polyphenols. Our results showed that 4,4′-di-O-methylellagic acid (4,4′-DiOMEA) was the most effective compound in the inhibition of colon cancer cell proliferation. 4,4′-DiOMEA was 13-fold more effective than other compounds of the same family. In addition, 4,4′-DiOMEA was very active against colon cancer cells resistant to the chemotherapeutic agent 5-fluoracil, whereas no effect was observed in nonmalignant colon cells. Moreover, no correlation between antiproliferative and antioxidant activities was found, further supporting that structure differences might result in dissimilar molecular targets involved in their differential effects. Finally, microarray analysis revealed that 4,4′-DiOMEA modulated Wnt signaling, which might be involved in the potential antitumor action of this compound. Our results suggest that structural-activity differences between EA and 4,4′-DiOMEA might constitute the basis for a new strategy in anticancer drug discovery based on these chemical modifications.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© American Society for Pharmacology and Experimental Therapeutics
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Attribution-NonCommercial-NoDerivatives 4.0 Internacional
The ellagic acid derivative 4,4′-Di-O-methylellagic acid efficiently inhibits colon cancer cell growth through a mechanism involving WNT16
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425882021-06-23T09:52:53Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Vargas, Teodoro
Moreno Rubio, Juan
Herranz, Jesús
Cejas, Paloma
Molina, Susana
González-Vallinas Garrachón, Margarita
Mendiola, Marta
Burgos, Emilio
Aguayo, Cristina
Custodio, Ana B.
Machado, Isidro
Ramos, David
Gironella, Meritxell
Espinosa Salinas, Isabel
Ramos, Ricardo
Martín Hernández, Roberto
Risueño, Alberto
Rivas, Javier de las
Reglero, Guillermo
Yaya, Ricardo
Fernández Martos, Carlos
Aparicio, Jorge
Maurel, Joan
Feliu, Jaime
Ramírez de Molina, Ana
2020-09-28T07:10:42Z
2020-09-28T07:10:42Z
2015
Oncotarget, 2015, vol. 6, n. 9, p. 7348-7363
1949-2553
http://uvadoc.uva.es/handle/10324/42588
10.18632/oncotarget.3130
7348
9
7363
Oncotarget
6
1949-2553
Lipid metabolism plays an essential role in carcinogenesis due to the requirements
of tumoral cells to sustain increased structural, energetic and biosynthetic precursor
demands for cell proliferation. We investigated the association between expression
of lipid metabolism-related genes and clinical outcome in intermediate-stage colon
cancer patients with the aim of identifying a metabolic profile associated with greater
malignancy and increased risk of relapse. Expression profile of 70 lipid metabolismrelated genes was determined in 77 patients with stage II colon cancer. Cox regression
analyses using c-index methodology was applied to identify a metabolic-related
signature associated to prognosis. The metabolic signature was further confirmed in
two independent validation sets of 120 patients and additionally, in a group of 264
patients from a public database. The combined analysis of these 4 genes, ABCA1,
ACSL1, AGPAT1 and SCD, constitutes a metabolic-signature (ColoLipidGene) able to
accurately stratify stage II colon cancer patients with 5-fold higher risk of relapse with
strong statistical power in the four independent groups of patients. The identification
www.impactjournals.com/oncotarget 7349 Oncotarget
of a group of 4 genes that predict survival in intermediate-stage colon cancer patients
allows delineation of a high-risk group that may benefit from adjuvant therapy, and
avoids the toxic and unnecessary chemotherapy in patients classified as low-risk group.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© Impact Journals
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
ColoLipidGene: signature of lipid metabolism-related genes to predict prognosis in stage-II colon cancer patients
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425892021-11-03T13:27:14Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Vargas, Teodoro
Moreno Rubio, Juan
Herranz, Jesús
Cejas, Paloma
Molina, Susana
González-Vallinas Garrachón, Margarita
Ramos, Ricardo
Burgos, Emilio
Aguayo, Cristina
Custodio, Ana B.
Reglero, Guillermo
Feliu, Jaime
Ramírez de Molina, Ana
2020-09-28T07:15:24Z
2020-09-28T07:15:24Z
2014
Molecular Oncology, 2014, vol. 8, n. 8, p.1469-1481
1574-7891
http://uvadoc.uva.es/handle/10324/42589
10.1016/j.molonc.2014.05.015
1469
8
1481
Molecular Oncology
8
Studies have recently suggested that metabolic syndrome and its components increase
the risk of colorectal cancer. Both diseases are increasing in most countries, and the genetic association between them has not been fully elucidated. The objective of this study
was to assess the association between genetic risk factors of metabolic syndrome or
related conditions (obesity, hyperlipidaemia, diabetes mellitus type 2) and clinical
outcome in stage II colorectal cancer patients. Expression levels of several genes related
to metabolic syndrome and associated alterations were analysed by real-time qPCR in
two equivalent but independent sets of stage II colorectal cancer patients. Using logistic
regression models and cross-validation analysis with all tumour samples, we developed
a metabolic syndrome-related gene expression profile to predict clinical outcome in
stage II colorectal cancer patients. The results showed that a gene expression profile
constituted by genes previously related to metabolic syndrome was significantly associated with clinical outcome of stage II colorectal cancer patients. This metabolic profile
was able to identify patients with a low risk and high risk of relapse. Its predictive value
was validated using an independent set of stage II colorectal cancer patients. The identification of a set of genes related to metabolic syndrome that predict survival in
intermediate-stage colorectal cancer patients allows delineation of a high-risk group
that may benefit from adjuvant therapy and avoid the toxic and unnecessary chemotherapy in patients classified as low risk. Our results also confirm the linkage between.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© Wiley Open Access
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Genes associated with metabolic syndrome predict disease-free survival in stage II colorectal cancer patients. A novel link between metabolic dysregulation and colorectal cancer
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/425902021-11-03T13:27:55Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
González-Vallinas Garrachón, Margarita
Molina, Susana
Vicente, Gonzalo
Zarza, Virginia
Martín Hernández, Roberto
García Risco, Mónica R.
Fornari, Tiziana
Reglero, Guillermo
Ramírez de Molina, Ana
2020-09-28T07:22:38Z
2020-09-28T07:22:38Z
2014
PLoS ONE, 2014, vol 9, n. 6, e98556
1932-6203
http://uvadoc.uva.es/handle/10324/42590
10.1371/journal.pone.0098556
e98556
6
PLoS ONE
9
1932-6203
Colorectal and pancreatic cancers remain important contributors to cancer mortality burden and, therefore, new
therapeutic approaches are urgently needed. Rosemary (Rosmarinus officinalis L.) extracts and its components have been
reported as natural potent antiproliferative agents against cancer cells. However, to potentially apply rosemary as a
complementary approach for cancer therapy, additional information regarding the most effective composition, its
antitumor effect in vivo and its main molecular mediators is still needed. In this work, five carnosic acid-rich supercritical
rosemary extracts with different chemical compositions have been assayed for their antitumor activity both in vivo (in nude
mice) and in vitro against colon and pancreatic cancer cells. We found that the antitumor effect of carnosic acid together
with carnosol was higher than the sum of their effects separately, which supports the use of the rosemary extract as a
whole. In addition, gene and microRNA expression analyses have been performed to ascertain its antitumor mechanism,
revealing that up-regulation of the metabolic-related gene GCNT3 and down-regulation of its potential epigenetic
modulator miR-15b correlate with the antitumor effect of rosemary. Moreover, plasmatic miR-15b down-regulation was
detected after in vivo treatment with rosemary. Our results support the use of carnosic acid-rich rosemary extract as a
complementary approach in colon and pancreatic cancer and indicate that GCNT3 expression may be involved in its
antitumor mechanism and that miR-15b might be used as a non-invasive biomarker to monitor rosemary anticancer effect.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© Public Library of Science
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Expression of MicroRNA-15b and the glycosyltransferase GCNT3 correlates with antitumor efficacy of rosemary diterpenes in colon and pancreatic cancer
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/427052021-06-23T09:52:58Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Vicente, Gonzalo
Molina, Susana
González-Vallinas Garrachón, Margarita
García Risco, Mónica R.
Fornari, Tiziana
Reglero, Guillermo
Ramírez de Molina, Ana
2020-10-01T09:39:02Z
2020-10-01T09:39:02Z
2013
The Journal of Supercritical Fluids, 2013, vol. 79, p. 101-108
0896-8446
http://uvadoc.uva.es/handle/10324/42705
10.1016/j.supflu.2012.07.006
101
108
The Journal of Supercritical Fluids
79
The use of supercritical fluid technology as an innovative technology to extract bioactive compounds has grown considerably in recent decades. Particularly, the recovery of antioxidants from different herbs is a matter of continuous research and development. Antioxidants can protect cells against the effects of free radicals and thus, play an important role in heart illness, cancer and other diseases.
Rosemary (Rosmarinus officinalis L.) has been recognized as one of the Lamiaceae plant with many important biological activities. Particularly, large antioxidant power has been recognized in rosemary and main substances related with this activity were the phenolic diterpenes such as carnosol, rosmanol, carnosic acid, methyl carnosate, and phenolic acids such as the rosmarinic and caffeic acids. Moreover, carnosic acid and carnosol are recognized as the most abundant antioxidants present in rosemary.
In this work, supercritical fluid technology was applied to produce rosemary extracts with different compositions and thus, with different bioactivity properties. Selected extracts, from the variety of samples obtained, were used to study the capability of rosemary supercritical extracts to inhibit the proliferation of human liver carcinoma cells. These extracts showed a dose-dependent effect on inhibiting the proliferation of human hepatoma cells. Moreover, observed citostaticity appeared to be significantly influenced by their different composition, suggesting a relevant role of the technology to produce the extracts and the consequently obtained compositions on the potential antitumoral activity of rosemary.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© Elsevier
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Supercritical rosemary extracts, their antioxidant activity and effect on hepatic tumor progression
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/444792021-06-24T07:36:56Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Cazaña Pérez, Violeta
Cidad Velasco, María del Pilar
Donate Correa, Javier
Martín Núñez, Ernesto
López López, José Ramón
Pérez García, María Teresa
Giráldez Fernández, Teresa
Navarro González, Juan Francisco
Álvarez de la Rosa, Diego
2020-12-21T12:59:12Z
2020-12-21T12:59:12Z
2018
Frontiers in Physiology, 2018, vol 9, n. 89. 14 p.
1664-042X
http://uvadoc.uva.es/handle/10324/44479
10.3389/fphys.2018.00089
Patients with chronic kidney disease (CKD) have a markedly increased incidence of cardiovascular disease (CVD). The high concentration of circulating uremic toxins and alterations in mineral metabolism and hormone levels produce vascular wall remodeling and significant vascular damage. Medial calcification is an early vascular event in CKD patients and is associated to apoptosis or necrosis and trans-differentiation of vascular smooth muscle cells (VSMC) to an osteogenic phenotype. VSMC obtained from bovine or rat aorta and cultured in the presence of increased inorganic phosphate (Pi) have been extensively used to study these processes. In this study we used human aortic VSMC primary cultures to compare the effects of increased Pi to treatment with serum obtained from uremic patients. Uremic serum induced calcification, trans-differentiation and phenotypic remodeling even with normal Pi levels. In spite of similar calcification kinetics, there were fundamental differences in osteochondrogenic marker expression and alkaline phosphatase induction between Pi and uremic serum-treated cells. Moreover, high Pi induced a dramatic decrease in cell viability, while uremic serum preserved it. In summary, our data suggests that primary cultures of human VSMC treated with serum from uremic patients provides a more informative model for the study of vascular calcification secondary to CKD.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2018 Frontiers
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Phenotypic modulation of cultured primary human aortic vascular smooth muscle cells by uremic serum
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/444802021-06-24T07:36:58Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Barrese, Vincenzo
Cidad Velasco, María del Pilar
Yeung, Shuk Y.
López López, José Ramón
McNeish, Alister J.
Ohya, Susumu
Pérez García, María Teresa
Greenwood, Iain A.
2020-12-21T13:18:01Z
2020-12-21T13:18:01Z
2017
Frontiers in Physiology, 2017, vol 8. 10 p.
1664-042X
http://uvadoc.uva.es/handle/10324/44480
10.3389/fphys.2017.00500
K+ channels encoded by the ether-a-go-go related gene (ERG1 or KCNH2) are important determinants of the cardiac action potential. Expression of both cardiac isoforms (ERG1a and ERG1b) were identified in murine portal vein and distinctive voltage-gated K+ currents were recorded from single myocytes. The aim of the present study was to ascertain the expression and functional impact of ERG channels in murine arteries.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2017 Frontiers
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Proliferative role of Kv11 channels in murine arteries
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/444822021-06-24T07:36:47Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Álvarez Miguel, Inés
Cidad Velasco, María del Pilar
Pérez García, María Teresa
López López, José Ramón
2020-12-21T13:58:55Z
2020-12-21T13:58:55Z
2016
The Journal of Physiology, 2017, vol. 595, n. 5. p. 1497-1513
1469-7793
http://uvadoc.uva.es/handle/10324/44482
10.1113/JP273327
Increased vascular tone in essential hypertension involves a sustained rise in total peripheral resistance. A model has been proposed in which the combination of membrane depolarization and higher L‐type Ca2+ channel activity generates augmented Ca2+ influx into vascular smooth muscle cells (VSMCs), contraction and vasoconstriction. The search for culprit ion channels responsible for membrane depolarization has provided several candidates, including members of the canonical transient receptor potential (TRPC) family. TRPC3 and TRPC6 are diacylglycerol‐activated, non‐selective cationic channels contributing to stretch‐ or agonist‐induced depolarization. Conflicting information exists regarding changes in TRPC3/TRPC6 functional expression in hypertension. However, although TRPC3‐TRPC6 channels can heteromultimerize, the possibility that differences in their association pattern may change their functional contribution to vascular tone is largely unexplored. We probe this hypothesis using a model of essential hypertension (BPH mice; blood pressure high) and its normotensive control (BPN mice; blood pressure normal). First, non‐selective cationic currents through homo‐ and heterotetramers recorded from transfected Chinese hamster ovary cells indicated that TRPC currents were sensitive to the selective antagonist Pyr10 only when TRPC6 was present, whereas intracellular anti‐TRPC3 antibody selectively blocked TRPC3‐mediated currents. In mesenteric VSMCs, basal and agonist‐induced currents were more sensitive to Pyr3 and Pyr10 in BPN cells. Consistently, myography studies showed a larger Pyr3/10‐induced vasodilatation in BPN mesenteric arteries. mRNA and protein expression data supported changes in TRPC3 and TRPC6 proportions and assembly, with a higher TRPC3 channel contribution in BPH VSMCs that could favour cell depolarization. These differences in functional and pharmacological properties of TRPC3 and TRPC6 channels, depending on their assembly, could represent novel therapeutical opportunities.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2016 The Physiological Society
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Differences in TRPC3 and TRPC6 channels assembly in mesenteric vascular smooth muscle cells in essential hypertension
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/445912021-06-24T07:36:50Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Cidad Velasco, María del Pilar
Novensá, Laura
Garabito, M.
Batlle, M.
Dantas, A. P.
Heras i Fortuny, Maria Magdalena
López López, José Ramón
Pérez García, María Teresa
Roqué, Mercé
2020-12-22T07:59:27Z
2020-12-22T07:59:27Z
2014
Cardiovascular Drugs and Therapy, 2014, vol. 28. p. 501-511
1573-7241
http://uvadoc.uva.es/handle/10324/44591
10.1007/s10557-014-6554-5
K+ channels are central to vascular pathophysiology. Previous results demonstrated that phenotypic modulation associates with a change in Kv1.3 to Kv1.5 expression, and that Kv1.3 blockade inhibits proliferation of VSMCs cultures.
Purpose: To explore whether the Kv1.3 to Kv1.5 switch could be a marker of the increased risk of intimal hyperplasia in essential hypertension and whether systemic treatment with Kv1.3 blockers can prevent intimal hyperplasia after endoluminal lesion . Methods: Morphometric and immunohistochemical analysis were performed in arterial segments following arterial injury and constant infusion of the Kv1.3 blocker PAP-1 during 28 days. Differential expression of K+ channel genes was studied in VSMC from hypertensive (BPH) and normotensive (BPN) mice, both in control and after endoluminal lesion. Finally, the migration and proliferation rate of BPN and BPH VSMCs was explored in vitro.
Results: Changes in mRNA expression led to an increased Kv1.3/Kv1.5 ratio in BPH VSMC. Consistent with this, arterial injury in BPH mice induced a higher degree of luminal stenosis, (84±4 % vs. 70±5 % in BPN, p<0.01), although no differences in migration and proliferation rate were observed in cultured VSMCs. The in vivo proliferative lesions were significantly decreased upon PAP-1 systemic infusion (18± 6 % vs. 58±20 % with vehicle, p<0.05).
Conclusions: Hypertension leads to a higher degree of luminal stenosis in our arterial injury model, that correlates with a decreased expression of Kv1.5 channels. Kv1.3 blockers decreased in vitro VSMCs proliferation, migration, and in vivo intimal hyperplasia formation, pointing to Kv1.3 channels as promising therapeutical targets against restenosis.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2014 Springer
Attribution-NonCommercial-NoDerivs 3.0 Unported
K+ channels expression in hypertension after arterial injury, and effect of selective Kv1.3 blockade with PAP-1 on intimal hyperplasia formation
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/445952021-06-24T07:36:49Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Tajada Esteban, Sendoa
Cidad Velasco, María del Pilar
Moreno Domínguez, Alejandro
Pérez García, María Teresa
López López, José Ramón
2020-12-22T09:16:41Z
2020-12-22T09:16:41Z
2012
The Journal of Physiology, 2012, vol. 590, n. 23. p. 6075-6091
1469-7793
http://uvadoc.uva.es/handle/10324/44595
10.1113/jphysiol.2012.236190
The increased vascular tone that defines essential hypertension is associated with depolarization of vascular smooth muscle cells (VSMCs) and involves a change in the expression profile of ion channels promoting arterial contraction. As a major regulator of VSMC resting membrane potential (VM), K+channel activity is an important determinant of vascular tone and vessel diameter. However, hypertension-associated changes in the expression and/or modulation of K+channels are poorly defined, due to their large molecular diversity and their bed-specific pattern of expression. Moreover, the impact of these changes on the integrated vessel functionand their contribution to the development of altered vascular tone under physiological conditions need to be confirmed. Hypertensive (BPH) and normotensive (BPN) mice strains obtained by phenotypic selection were used to explore whether changes in the functional expression of VSMC inward rectifier K+channels contribute to the more depolarized resting VM and the increased vascular reactivity of hypertensive arteries. We determined the expression levels of inward rectifierK+channel mRNA in several vascular beds from BPN and BPH animals, and their functional contribution to VSMC excitability and vascular tone in mesenteric arteries. We found a decrease in the expression of Kir2.1, Kir4.1, Kir6.x and SUR2 mRNA in BPH VSMCs, and a decreased functional contribution of both KIRand KATP channels in isolated BPH VSMCs. However, only
the effect of KATP channel modulators was impaired when exploring vascular tone, suggesting that decreased functional expression of KATP channels may be an important element in the remodelling of VSMCs in essential hypertension.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2012 The Journal of Physiology
Attribution-NonCommercial-NoDerivs 3.0 Unported
High blood pressure associates with the remodelling of inward rectifier K+ channels in mice mesenteric vascular smooth muscle cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/445972021-06-24T07:36:43Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Miguel Velado, Eduardo
Pérez Carretero, Francisco D.
Colinas, Olaia
Cidad Velasco, María del Pilar
Heras i Fortuny, Maria Magdalena
López López, José Ramón
Pérez García, María Teresa
2020-12-22T09:48:51Z
2020-12-22T09:48:51Z
2010
Cardiovascular Research, 2010, vol. 86, n. 3. p. 383-391
1755-3245
http://uvadoc.uva.es/handle/10324/44597
10.1093/cvr/cvq011
Aims: Vascular smooth muscle cell (VSMC) proliferation is involved in cardiovascular pathologies associated with unwanted arterial wall remodelling. Coordinated changes in the expression of several K+ channels have been found to be important elements in the phenotypic switch of VSMCs towards proliferation. We have previously demonstrated the association of functional expression of Kv3.4 channels with proliferation of human uterine VSMCs. Here, we sought to gain deeper insight on the relationship between Kv3.4 channels and cell cycle progression in this preparation. Methods and results: Expression and function of Kv3.4 channels along the cell cycle was explored in uterine VSMCs synchronized at different checkpoints, combining real-time PCR, western blotting, and electrophysiological techniques. Flow cytometry, Ki67 expression and BrdU incorporation techniques allowed us to explore the effects of Kv3.4 channels blockade on cell cycle distribution. We found cyclic changes in Kv3.4 and MiRP2 mRNA and protein expression along the cell cycle. Functional studies showed that Kv3.4 current amplitude and Kv3.4 channels contribution to cell excitability increased in proliferating cells. Finally, both Kv3.4 blockers and Kv3.4 knockdown with siRNA reduced the proportion of proliferating VSMCs. Conclusion: Our data indicate that Kv3.4 channels exert a permissive role in the cell cycle progression of proliferating uterine VSMCs, as their blockade induces cell cycle arrest after G2/M phase completion. The modulation of resting membrane potential (VM) by Kv3.4 channels in proliferating VSMCs suggests that their role in cell cycle progression could be at least in part mediated by their contribution to the hyperpolarizing signal needed to progress through the G1 phase.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2010 Oxford University Press
Attribution-NonCommercial-NoDerivs 3.0 Unported
Cell cycle-dependent expression of Kv3.4 channels modulates proliferation of human uterine artery smooth muscle cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/445992021-06-24T07:36:44Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Cidad Velasco, María del Pilar
Moreno Domínguez, Alejandro
Novensá, Laura
Roqué, Mercé
Barquín, Leire
Heras i Fortuny, Maria Magdalena
Pérez García, María Teresa
López López, José Ramón
2020-12-22T10:25:28Z
2020-12-22T10:25:28Z
2010
Arteriosclerosis, Thrombosis, and Vascular Biology, 2010, vol. 30, n. 6. p. 1203-1211
1524-4636
http://uvadoc.uva.es/handle/10324/44599
10.1161/ATVBAHA.110.205187
Objective: Vascular smooth muscle cells (VSMCs) contribute significantly to occlusive vascular diseases by virtue of their ability to switch to a noncontractile, migratory, and proliferating phenotype. Although the participation of ion channels in this phenotypic modulation (PM) has been described previously, changes in their expression are poorly defined because of their large molecular diversity. We obtained a global portrait of ion channel expression in contractile versus proliferating mouse femoral artery VSMCs, and explored the functional contribution to the PM of the most relevant changes that we observed.
Methods and Results: High-throughput real-time polymerase chain reaction of 87 ion channel genes was performed in 2 experimental paradigms: an in vivo model of endoluminal lesion and an in vitro model of cultured VSMCs obtained from explants. mRNA expression changes showed a good correlation between the 2 proliferative models, with only 2 genes, Kv1.3 and Kvβ2, increasing their expression on proliferation. The functional characterization demonstrates that Kv1.3 currents increased in proliferating VSMC and that their selective blockade inhibits migration and proliferation.
Conclusion: These findings establish the involvement of Kv1.3 channels in the PM of VSMCs, providing a new therapeutical target for the treatment of intimal hyperplasia.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2010 American Heart Association
Attribution-NonCommercial-NoDerivs 3.0 Unported
Characterization of ion channels involved in the proliferative response of femoral artery smooth muscle cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/446002021-06-24T07:36:46Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Miguel Velado, Eduardo
Moreno Domínguez, Alejandro
Colinas, Olaia
Cidad Velasco, María del Pilar
Heras i Fortuny, Maria Magdalena
Pérez García, María Teresa
López López, José Ramón
2020-12-22T10:56:25Z
2020-12-22T10:56:25Z
2005
Circulation Research, 2005, vol. 97, n. 12. p. 1280-1287
1524-4571
http://uvadoc.uva.es/handle/10324/44600
10.1161/01.RES.0000194322.91255.13
Vascular smooth muscle cells (VSMCs) perform diverse functions that can be classified into contractile and synthetic (or proliferating). All of these functions can be fulfilled by the same cell because of its capacity of phenotypic modulation in response to environmental changes. The resting membrane potential is a key determinant for both contractile and proliferating functions. Here, we have explored the expression of voltage-dependent K+ (Kv) channels in contractile (freshly dissociated) and proliferating (cultured) VSMCs obtained from human uterine arteries to establish their contribution to the functional properties of the cells and their possible participation in the phenotypic switch. We have studied the expression pattern (both at the mRNA and at the protein level) of Kvα subunits in both preparations as well as their functional contribution to the K+ currents of VSMCs. Our results indicate that phenotypic remodeling associates with a change in the expression and distribution of Kv channels. Whereas Kv currents in contractile VSMCs are mainly performed by Kv1 channels, Kv3.4 is the principal contributor to K+ currents in cultured VSMCs. Furthermore, selective blockade of Kv3.4 channels resulted in a reduced proliferation rate, suggesting a link between Kv channels expression and phenotypic remodeling.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2005 American Heart Association
Attribution-NonCommercial-NoDerivs 3.0 Unported
Contribution of Kv channels to phenotypic remodeling of human uterine artery smooth muscle cells
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/446022021-06-24T07:36:40Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
López López, José Ramón
Pérez García, María Teresa
2020-12-22T12:24:21Z
2020-12-22T12:24:21Z
2007
Circulation Research, 2007, vol. 101, n. 10. p. 965-967
1524-4571
http://uvadoc.uva.es/handle/10324/44602
10.1161/CIRCRESAHA.107.164442
Spontaneous breathing requires feedback controls in which detection of blood gas and pH are critical. While O2 detection is performed by pheripheral chemoreceptors, CO2/pH-sensitive chemoreceptors are in the carotid bodies (CBs), but major sites are also within the brain (the central chemoreceptors [CCR]). CO2/pH signals are related to the acid-base status of the blood and reflect the adequacy of breathing to metabolism. Small changes in CO2/pH can affect breathing, so that a rise in Pco2 as small as 1 mm Hg produces an evident change in ventilation.1 Such a high CO2 sensitivity relies in the inherent properties of CO2/pH-sensing molecules present both in CB and CCR cells, as shown in several recent studies on CO2/pH sensing ion channels and receptors.2,3 Functional properties of most proteins can be regulated by changes in pH, as this would only require 1 or a few titratable residues of the molecule, whose protonation can lead to conformation changes that translate into changes in activity. However, to define whether those molecules have a relevant role in CO2/pH chemoreception, some more criteria should be met, including their range of pH sensitivity, their location in chemoreceptor cells, and their functional contribution to the integrated chemoreceptor response. In this regard, the molecular characterization of pH-sensitive channels and transporters has progressed considerably within the last years, but conclusive evidences of their contribution to acid chemotransduction are not so well established for many of them. The main reason for this delay is the fact that the identity of the primary sensory cells constituting CCRs and of their neuronal networks remains elusive. In vitro, neurons from many brain locations are excited or inhibited by CO2/pH changes, but it has been difficult to link this neuronal chemosensitivity to chemoreception in vivo. Moreover, in addition to chemosensitive neurons, CCRs sites may also contain neurons with other integrative functions that are not clearly distinct from chemosensitive ones in their morphological or functional properties, making their experimental study difficult.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2007 American Heart Association
Attribution-NonCommercial-NoDerivs 3.0 Unported
An ASIC channel for acid chemotransduction
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/446052021-10-18T06:33:48Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Riesco Fagundo, Armenia María
Pérez García, María Teresa
González, C.
López López, José Ramón
2020-12-22T14:02:53Z
2020-12-22T14:02:53Z
2001
Circulation Research, 2001, vol. 89, n. 5. p. 430-436
1524-4571
http://uvadoc.uva.es/handle/10324/44605
10.1161/hh1701.095632
Hypoxic inhibition of large-conductance Ca2+-dependent K+ channels (maxiK) of rat carotid body type I cells is a well-established fact. However, the molecular mechanisms of such inhibition and the role of these channels in the process of hypoxic transduction remain unclear. We have examined the mechanisms of interaction of O2 with maxiK channels exploring the effect of hypoxia on maxiK currents recorded with the whole-cell and the inside-out configuration of the patch-clamp technique. Hypoxia inhibits channel activity both in whole-cell and in excised membrane patches. This effect is strongly voltage- and Ca2+-dependent, being maximal at low [Ca2+] and low membrane potential. The analysis of single-channel kinetics reveals a gating scheme comprising three open and five closed states. Hypoxia inhibits channel activity increasing the time the channel spends in the longest closed states, an effect that could be explained by a decrease in the Ca2+ sensitivity of those closed states. Reducing maxiK channels with dithiothreitol (DTT) increases channel open probability, whereas oxidizing the channels with 2,2′-dithiopyridine (DTDP) has the opposite effect. These results suggest that hypoxic inhibition is not related with a reduction of channel thiol groups. However, CO, a competitive inhibitor of O2 binding to hemoproteins, fully reverts hypoxic inhibition, both at the whole-cell and the single-channel level. We conclude that O2 interaction with maxiK channels does not require cytoplasmic mediators. Such interaction could be mediated by a membrane hemoprotein that, as an O2 sensor, would modulate channel activity.
eng
info:eu-repo/semantics/openAccess
© 2001 American Heart Association
Hypoxia
Hipoxia
O2 modulates large-conductance Ca2+-dependent K+ channels of rat chemoreceptor cells by a membrane-restricted and CO-sensitive mechanism
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/446062021-06-24T07:36:42Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Pérez García, María Teresa
López López, José Ramón
2020-12-28T12:48:36Z
2020-12-28T12:48:36Z
2000
Circulation Research, 2000, vol. 86, n. 5. p. 490-491
1524-4571
http://uvadoc.uva.es/handle/10324/44606
10.1161/01.RES.86.5.490
This report represents a relevant contribution to the study of oxygen sensing for two main reasons. First, it shows an approach adequate for identification of a putative O2-sensitive K+ channel, by moving from the modulation by hypoxia of a recombinant channel back to its physiological context, the role of this channel in the response to low Po2 of the native cells. Second, it demonstrates that hypoxic inhibition of the recombinant Kv3.1 channels is retained in excised membrane patches, pointing to a membrane-delimited mechanism as the origin of hypoxic responses. The importance of this latter finding deserves additional comment.
eng
info:eu-repo/semantics/openAccess
© 2000 American Heart Association
Are Kv channels the essence of O2 sensing?
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/446072021-06-24T07:36:52Zcom_10324_1134com_10324_931com_10324_894com_10324_32522com_10324_952com_10324_43677com_10324_954col_10324_1213col_10324_32523col_10324_43678
Cidad Velasco, María del Pilar
Jiménez Pérez, Laura
García Arribas, Daniel
Miguel Velado, Eduardo
Tajada Esteban, Sendoa
Ruiz McDavitt, Christian
López López, José Ramón
Pérez García, María Teresa
2020-12-28T13:15:13Z
2020-12-28T13:15:13Z
2012
Arteriosclerosis, Thrombosis, and Vascular Biology, 2012, vol. 32, n. 5. p. 1299-1307
1524-4636
http://uvadoc.uva.es/handle/10324/44607
10.1161/ATVBAHA.111.242727
Objective: Phenotypic modulation of vascular smooth muscle cells has been associated with a decreased expression of all
voltage-dependent potassium channel (Kv)1 channel encoding genes but Kcna3 (which encodes Kv1.3 channels). In fact,
upregulation of Kv1.3 currents seems to be important to modulate proliferation of mice femoral vascular smooth muscle
cells in culture. This study was designed to explore if these changes in Kv1 expression pattern constituted a landmark of
phenotypic modulation across vascular beds and to investigate the mechanisms involved in the proproliferative function
of Kv1.3 channels.
Methods and Results: Changes in Kv1.3 and Kv1.5 channel expression were reproduced in mesenteric and aortic vascular
smooth muscle cells, and their correlate with protein expression was electrophysiologicaly confirmed using selective
blockers. Heterologous expression of Kv1.3 and Kv1.5 channels in HEK cells has opposite effects on the proliferation
rate. The proproliferative effect of Kv1.3 channels was reproduced by “poreless” mutants but disappeared when voltagedependence of gating was suppressed.
Conclusion: These findings suggest that the signaling cascade linking Kv1.3 functional expression to cell proliferation is
activated by the voltage-dependent conformational change of the channels without needing ion conduction. Additionally,
the conserved upregulation of Kv1.3 on phenotypic modulation in several vascular beds makes this channel a good target
to control unwanted vascular remodeling.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/
© 2012 American Heart Association
Attribution-NonCommercial-NoDerivs 3.0 Unported
Kv1.3 channels can modulate cell proliferation during phenotypic switch by an ion-flux independent mechanism
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/469622021-06-23T09:53:19Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ribeiro, Maria J.
Sacramento, Joana F.
Gallego Martín, Teresa
Olea Fraile, Elena
Melo, Bernardete F.
Guarino, Maria P.
Yubero Benito, Sara
Obeso Cáceres, Ana María de la Luz
Conde, Silvia V.
2021-06-21T09:51:56Z
2021-06-21T09:51:56Z
2018
The Journal of Physiology, 2018, vol. 596, n. 15. p. 3187-3199
1469-7793
https://uvadoc.uva.es/handle/10324/46962
10.1113/JP275362
Leptin plays a role in the control of breathing, acting mainly on central nervous system structures. Leptin receptors are expressed in the carotid body (CB) and this finding has been associated with a putative physiological role of leptin in the regulation of CB function. Since, the CBs are implicated in energy metabolism, here we tested the effects of different concentrations of leptin administration on ventilatory parameters and on carotid sinus nerve (CSN) activity in control and high-fat (HF) diet fed rats, in order to clarify the role of leptin in ventilation control in metabolic disease states. We also investigated the expression of leptin receptors and the neurotransmitters involved in leptin signalling in the CBs. We found that in non-disease conditions, leptin increases minute ventilation in both basal and hypoxic conditions. However, in the HF model, the effect of leptin in ventilatory control is blunted. We also observed that HF rats display an increased frequency of CSN discharge in basal conditions that is not altered by leptin, in contrast to what is observed in control animals. Leptin did not modify intracellular Ca2+ in CB chemoreceptor cells, but it produced an increase in the release of adenosine from the whole CB. We conclude that CBs represent an important target for leptin signalling, not only to coordinate peripheral ventilatory chemoreflexive drive, but probably also to modulate metabolic variables. We also concluded that leptin signalling is mediated by adenosine release and that HF diets blunt leptin responses in the CB, compromising ventilatory adaptation.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2018 The Physiological Society
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
High fat diet blunts the effects of leptin on ventilation and on carotid body activity
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/469632021-06-23T09:53:21Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Quintero Coca, Miguel
Olea Fraile, Elena
Conde, Silvia V.
Gallego Martín, Teresa
González, C.
González, Constancio
Montserrat, Josep M.
Gómez Niño, María Ángeles
Yubero Benito, Sara
Agapito Serrano, María Teresa
2021-06-21T10:12:40Z
2021-06-21T10:12:40Z
2016
The Journal of Physiology, 2016, vol. 594, n. 6. p. 1773–1790
1469-7793
https://uvadoc.uva.es/handle/10324/46963
10.1113/JP270878
Obstructive sleep apnoea (OSA) affects an estimated 3–7% of the adult population, the frequency doubling at ages >60–65 years. As it evolves, OSA becomes frequently associated with cardiovascular, metabolic and neuropsychiatric pathologies defining OSA syndrome (OSAS). Exposing experimental animals to chronic intermittent hypoxia (CIH) can be used as a model of the recurrent hypoxic and O2 desaturation patterns observed in OSA patients. CIH is an important OSA event triggering associated pathologies; CIH induces carotid body (CB)-driven exaggerated sympathetic tone and overproduction of reactive oxygen species, related to the pathogenic mechanisms of associated pathologies observed in OSAS. Aiming to discover why OSAS is clinically less conspicuous in aged patients, the present study compares CIH effects in young (3–4 months) and aged (22–24 months) rats. To define potential distinctive patterns of these pathogenic mechanisms, mean arterial blood pressure as the final CIH outcome was measured. In young rats, CIH augmented CB sensory responses to hypoxia, decreased hypoxic ventilation and augmented sympathetic activity (plasma catecholamine levels and renal artery content and synthesis rate). An increased brainstem integration of CB sensory input as a trigger of sympathetic activity is suggested. CIH also caused an oxidative status decreasing aconitase/fumarase ratio and superoxide dismutase activity. In aged animals, CIH minimally affected CB responses, ventilation and sympathetic-related parameters leaving redox status unaltered. In young animals, CIH caused hypertension and in aged animals, whose baseline blood pressure was augmented, CIH did not augment it further. Plausible mechanisms of the differences and potential significance of these findings for the diagnosis and therapy of OSAS are discussed.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2016 The Physiological Society
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Age protects from harmful effects produced by chronic intermittent hypoxia
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/469662021-06-23T09:53:22Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Prieto Lloret, Jesús
Ramírez Arroyo, María
Olea Fraile, Elena
Moral Sanz, Javier
Cogolludo Torralba, Ángel
Castañeda, Javier
Yubero Benito, Sara
Agapito Serrano, María Teresa
Gómez Niño, María Ángeles
Rocher Martín, María Asunción
Rigual Bonastre, Ricardo Jaime
Obeso Cáceres, Ana María de la Luz
Pérez Vizcaíno, Francisco
González, Constancio
2021-06-21T11:25:11Z
2021-06-21T11:25:11Z
2015
The Journal of Physiology, 2015, vol. 593, n. 11. p. 2459-2477
1469-7793
https://uvadoc.uva.es/handle/10324/46966
10.1113/JP270274
Adult mammalians possess three cell systems that are activated by acute bodily hypoxia: pulmonary artery smooth muscle cells (PASMC), carotid body chemoreceptor cells (CBCC) and erythropoietin (EPO)-producing cells. In rats, chronic perinatal hyperoxia causes permanent carotid body (CB) atrophy and functional alterations of surviving CBCC. There are no studies on PASMC or EPO-producing cells. Our aim is to define possible long-lasting functional changes in PASMC or EPO-producing cells (measured as EPO plasma levels) and, further, to analyse CBCC functional alterations. We used 3- to 4-month-old rats born and reared in a normal atmosphere or exposed to perinatal hyperoxia (55–60% O2 for the last 5–6 days of pregnancy and 4 weeks after birth). Perinatal hyperoxia causes an almost complete loss of hypoxic pulmonary vasoconstriction (HPV), which was correlated with lung oxidative status in early postnatal life and prevented by antioxidant supplementation in the diet. O2-sensitivity of K+ currents in the PASMC of hyperoxic animals is normal, indicating that their inhibition is not sufficient to trigger HPV. Perinatal hyperoxia also abrogated responses elicited by hypoxia on catecholamine and cAMP metabolism in the CB. An increase in EPO plasma levels elicited by hypoxia was identical in hyperoxic and control animals, implying a normal functioning of EPO-producing cells. The loss of HPV observed in adult rats and caused by perinatal hyperoxia, comparable to oxygen therapy in premature infants, might represent a previously unrecognized complication of such a medical intervention capable of aggravating medical conditions such as regional pneumonias, atelectases or general anaesthesia in adult life.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2015 The Journal of Physiology
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Hypoxic pulmonary vasoconstriction, carotid body function and erythropoietin production in adult rats perinatally exposed to hyperoxia
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/469682021-06-23T09:53:23Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Yubero Benito, Sara
Ramudo, Laura
Manso, Manuel Antonio
Dios, Isabel de
2021-06-21T11:54:57Z
2021-06-21T11:54:57Z
2009
Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2009, vol. 1792, n. 12. p. 1205-1211
0925-4439
https://uvadoc.uva.es/handle/10324/46968
10.1016/j.bbadis.2009.10.001
This study aimed to investigate the role of therapeutic dexamethasone (Dex) treatment on the mechanisms underlying chemokine expression during mild and severe acute pancreatitis (AP) experimentally induced in rats. Regardless of the AP severity, Dex (1 mg/kg), administered 1 h after AP, reduced the acinar cell activation of extracellular signal-regulated kinase (ERK) and c-Jun-NH2-terminal kinase (JNK) but failed to reduce p38-mitogen-activated protein kinase (MAPK) in severe AP. In both AP models, Dex inhibited the activation of nuclear factor-kappaB (NF-κB) and signal transducers and activators of transcription (STAT) factors. All of this resulted in pancreatic down-regulation of the chemokines monocyte chemoattractant protein-1 (MCP-1) and cytokine-induced neutrophil chemoattractant (CINC). Lower plasma chemokine levels as well as decreased amylasemia, hematocrit and plasma interleukin-1β (Il-1β) levels were found either in mild or severe AP treated with Dex. Pancreatic neutrophil infiltration was attenuated by Dex in mild but not in severe AP. In conclusion, by targeting MAPKs, NF-κB and STAT3 pathways, Dex treatment down-regulated the chemokine expression in different cell sources during mild and severe AP, resulting in decreased severity of the disease.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2009 Elsevier
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Mechanisms of dexamethasone-mediated chemokine down-regulation in mild and severe acute pancreatitis
info:eu-repo/semantics/article
oai:uvadoc.uva.es:10324/469702021-06-23T09:53:25Zcom_10324_1134com_10324_931com_10324_894col_10324_1213
Ramudo, Laura
Yubero Benito, Sara
Manso, Manuel Antonio
Vicente, Secundino
Dios, Isabel de
2021-06-21T12:24:37Z
2021-06-21T12:24:37Z
2009
Journal of Cellular and Molecular Medicine, 2009, vol. 13, n. 7. p. 1314-1320
1582-4934
https://uvadoc.uva.es/handle/10324/46970
10.1111/j.1582-4934.2008.00529.x
Pancreatitis-associated ascitic fluid (PAAF) is known to contribute to the progression of acute pancreatitis (AP). We have investigated the capability of PAAF to activate the expression of MCP-1 in pancreatic acinar cells and the involvement of MAPK, NF-κB and STAT3 as downstream signalling transduction pathways. The actions of dexamethasone (Dx) and N-acetylcysteine (NAC) on the PAAF's acinar effects have also been evaluated. Acinar cells were incubated for 1 hr with PAAF collected from rats with severe AP induced by sodium taurocholate in the absence or presence of Dx (10−7 M) or NAC (30 mM). MCP-1 mRNA expression, phospho-p38-MAPK, IκBα, nuclear p65 levels and nuclear translocation of STAT3 were analysed. In response to PAAF, overexpression of MCP-1, phosphorylation of p38-MAPK, degradation of IκBα and increases in p65 nuclear levels and STAT3 activity were found in acinar cells. PAAF-mediated MCP-1 up-regulation was completely suppressed by Dx and NAC. MAPK activation was only inhibited by NAC, NF-κB activation was repressed by Dx and NAC, and STAT3 pathway was strongly blocked by Dx and significantly reduced by NAC. In conclusion, acinar cells were activated by PAAF to produce MCP-1, mainly via NF-κB and STAT3 pathways. Both downstream pathways were targeted by Dx and NAC to repress the PAAF-mediated acinar MCP-1 up-regulation.
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
© 2009 Wiley
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Signal transduction of MCP-1 expression induced by pancreatitis-associated ascitic fluid in pancreatic acinar cells
info:eu-repo/semantics/article
mods///col_10324_1213/100