Electrophysiological characterization of store-operated and agonist-induced Ca2+ entry pathways in endothelial cells

Girardin, Nathalie; Antigny, Fabrice; Frieden, Maud

doi:10.1007/s00424-010-0825-1

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Title		Electrophysiological characterization of store-operated and agonist-induced Ca2+ entry pathways in endothelial cells
Authors		Girardin, Nathalie Antigny, Fabrice Frieden, Maud
Published in		Pflügers Archiv. 2010, vol. 460, no. 1, p. 109-120
Collection		Open Access - Licence nationale Springer
Abstract		In endothelial cells, agonist-induced Ca(2+) entry takes place via the store-operated Ca(2+) entry pathway and/or via channel(s) gated by second messengers. As cell stimulation leads to both a partial Ca(2+) store depletion as well as the production of second messengers, these two pathways are problematic to distinguish. We showed that passive endoplasmic reticulum (ER) depletion by thapsigargin or cell stimulation by histamine activated a similar Ca(2+)-release-activated Ca(2+) current (CRAC)-like current when 10 mM Ba(2+)/2 mM Ca(2+) was present in the extracellular solution. Importantly, during voltage clamp recordings, histamine stimulation largely depleted the ER Ca(2+) store, explaining the activation of a CRAC-like current (due to store depletion) upon histamine in Ba(2+) medium. On the contrary, in the presence of 10 mM Ca(2+), the ER Ca(2+) content remained elevated, and histamine induced an outward rectifying current that was inhibited by Ni(2+) and KB-R7943, two blockers of the Na(+)/Ca(2+) exchanger (NCX). Both blockers also reduced histamine-induced cytosolic Ca(2+) elevation. In addition, removing extracellular Na(+) increased the current amplitude which is in line with a current supported by the NCX. These data are consistent with the involvement of the NCX working in reverse mode (Na(+) out/Ca(2+) in) during agonist-induced Ca(2+) entry in endothelial cells.
Keywords		Barium/metabolism — Calcium/metabolism — Calcium Channels/drug effects/metabolism — Calcium Signaling/drug effects — Calcium-Transporting ATPases/antagonists & inhibitors/metabolism — Cell Line — Endoplasmic Reticulum/drug effects/metabolism — Endothelial Cells/metabolism — Enzyme Inhibitors/pharmacology — Histamine/pharmacology — Humans — Membrane Potentials — Nickel/metabolism — Patch-Clamp Techniques — Sodium/metabolism — Sodium-Calcium Exchanger/antagonists & inhibitors/metabolism — Thapsigargin/pharmacology — Thiourea/analogs & derivatives/pharmacology — Time Factors — Transfection
Identifiers		DOI: 10.1007/s00424-010-0825-1 PMID: 20419508
Full text		Article (Published version) - Free access Other version: http://www.springerlink.com/content/j7677h3371247386/fulltext.pdf
Structures		Faculté de médecine / Section de médecine fondamentale / Département de physiologie cellulaire et métabolisme
Research groups		Canaux ioniques endothéliaux (828) Signaux intracellulaires (210)
Citation (ISO format)		GIRARDIN, Nathalie, ANTIGNY, Fabrice, FRIEDEN, Maud. Electrophysiological characterization of store-operated and agonist-induced Ca2+ entry pathways in endothelial cells. In: Pflügers Archiv, 2010, vol. 460, n° 1, p. 109-120. doi: 10.1007/s00424-010-0825-1 https://archive-ouverte.unige.ch/unige:20967