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    • Dpto. Biología Celular, Genética, Histología y Farmacología
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    Por favor, use este identificador para citar o enlazar este ítem:http://uvadoc.uva.es/handle/10324/5950

    Título
    Functional measurements of [Ca2+] in the endoplasmic reticulum using a herpes virus to deliver targeted aequorin
    Autor
    Alonso Alonso, María TeresaAutoridad UVA
    Barrero, María José
    Carnicero Gila, Estela MaríaAutoridad UVA Orcid
    Montero Zoccola, María TeresaAutoridad UVA Orcid
    García-Sancho Martín, Francisco JavierAutoridad UVA Orcid
    Álvarez Martín, JavierAutoridad UVA Orcid
    Año del Documento
    1998
    Editorial
    Harcourt Brace & Co. Ltd
    Descripción
    Producción Científica
    Documento Fuente
    Cell Calcium, 1998, vol. 24, n. 2, p. 87-96
    Resumen
    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.
    Materias (normalizadas)
    Calcio - Metabolismo
    ISSN
    0143-4160
    Revisión por pares
    SI
    Idioma
    eng
    URI
    http://uvadoc.uva.es/handle/10324/5950
    Derechos
    openAccess
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    • DEP05 - Artículos de revista [198]
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    Universidad de Valladolid

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