RT info:eu-repo/semantics/article
T1 Functional measurements of [Ca2+] in the endoplasmic reticulum using a herpes virus to deliver targeted aequorin
A1 Alonso Alonso, María Teresa
A1 Barrero, María José
A1 Carnicero Gila, Estela María
A1 Montero Zoccola, María Teresa
A1 García-Sancho Martín, Francisco Javier
A1 Álvarez Martín, Javier
K1 Calcio - Metabolismo
AB Changes in the free calcium concentration of the endoplasmic reticulum ([Ca 2+],,) play a central rolecontrolling cellular functions like contraction, secretion or neuronal signaling. We recently reported that recombinantaequorin targeted to the endoplasmic reticulum (ER) [Montero M., Brini M., Marsault R. et al. Monitoring dynamicchanges 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 J1997; 11: 881-8861 can be used to monitor selectively [Ca2+le, in intact HeLa cells. Here we have used a herpessimplex virus type 1 (HSV-1) based system to deliver targeted aequorin into a number of different cell types includingboth 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 around300 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 allthe excitable cells (anterior pituitary, GH,, chromaffin cells and granule neurons). We conclude that delivery ofrecombinant aequorin to the ER using HSV amplicon provides the first direct quantitative and dynamic measurementsof [Ca2+le, in several primary non-dividing cells.
PB Harcourt Brace & Co. Ltd
SN 0143-4160
YR 1998
FD 1998
LK http://uvadoc.uva.es/handle/10324/5950
UL http://uvadoc.uva.es/handle/10324/5950
LA eng
NO Cell Calcium, 1998, vol. 24, n. 2, p. 87-96
NO Producción Científica
DS UVaDOC
RD 26-abr-2024