2024-03-29T01:50:13Zhttps://uvadoc.uva.es/oai/requestoai:uvadoc.uva.es:10324/59502021-06-23T09:48:17Zcom_10324_1133com_10324_931com_10324_894col_10324_1209
Functional measurements of [Ca2+] in the endoplasmic reticulum using a herpes virus to deliver targeted aequorin
Alonso Alonso, María Teresa
Barrero, María José
Carnicero Gila, Estela María
Montero Zoccola, María Teresa
García-Sancho Martín, Francisco Javier
Álvarez Martín, Javier
Calcio - Metabolismo
Changes in the free calcium concentration of the endoplasmic reticulum ([Ca 2+],,) play a central role
controlling cellular functions like contraction, secretion or neuronal signaling. We recently reported that recombinant
aequorin targeted to the endoplasmic reticulum (ER) [Montero M., Brini M., Marsault R. et al. Monitoring dynamic
changes in free Ca2+ concentration in the endoplasmic reticulum of intact cells. EMBO J 1995; 14: 5467-5475,
Montero M., Barrero M.J., Alvarez J. [Ca2+] microdomains control agonist-induced Ca2+ release in intact cells. FASEB J
1997; 11: 881-8861 can be used to monitor selectively [Ca2+le, in intact HeLa cells. Here we have used a herpes
simplex virus type 1 (HSV-1) based system to deliver targeted aequorin into a number of different cell types including
both postmitotic primary cells (anterior pituitary cells, chromaffin cells and cerebellar neurons) and cell lines (HeLa,
NIH3T3, GH, and PC12 cells). Functional studies showed that the steady state lumenal [Ca*+],, ranged from around
300 pM in granule cells to 800 ).rM in GH,cells. InsP,-coupled receptor stimulation with agonists like histamine (in HeLa,
NIH3T3 and chromaffin cells), UTP and bradykinin (in PC12 cells) or thyrotropin-releasing hormone (TRH, in GH,cells)
produced a very rapid decrease in lumenal [Ca’+],,. Caffeine caused a rapid Ca2+ depletion of the ER in chromaffin cells,
but not in the other cell types. Depolarization by high K+ produced an immediate and reversible increase of [Ca2+lerin all
the excitable cells (anterior pituitary, GH,, chromaffin cells and granule neurons). We conclude that delivery of
recombinant aequorin to the ER using HSV amplicon provides the first direct quantitative and dynamic measurements
of [Ca2+le, in several primary non-dividing cells.
2014-09-15T11:18:03Z
2014-09-15T11:18:03Z
2014-09-15T11:18:03Z
1998
info:eu-repo/semantics/article
Cell Calcium, 1998, vol. 24, n. 2, p. 87-96
0143-4160
http://uvadoc.uva.es/handle/10324/5950
87
2
96
Cell Calcium
24
eng
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Attribution-NonCommercial-NoDerivatives 4.0 International
Harcourt Brace & Co. Ltd