title: Calcium influx into endothelial cells and formation of endothelium-derived relaxing factor is controlled by the membrane potential.
abstract: We studied the role of the membrane potential in the control of the intracellular free calcium concentration ([Ca2+]i) and release of the two autacoids endothelium-derived relaxing factor (EDRF = nitric oxide) and prostaglandin I2 in endothelial cells. ATP (3 mumol/l) and bradykinin (1 nmol/l) evoked rapid increases (sixfold) in [Ca2+]i in cultured endothelial cells. [Ca2+]i remained elevated over several minutes. When the cells were depolarized, either by K+ (70-90 mmol/l) or by preincubation with the blocker of K+ channels tetraethylammonium (3 mmol/l), the initial peak of [Ca2+]i remained unaffected but [Ca2+]i returned significantly faster to resting levels, indicating a reduction in Ca2+ influx. In native, freshly isolated endothelial cells, K+ abolished increases in [Ca2+]i induced by acetylcholine (3 mumol/l). Release of EDRF in response to bradykinin (cultured cells) and acetylcholine (native cells) was inhibited by K+ (by 70%), whereas release of prostaglandin I2 was not significantly reduced. Preincubation of cultured endothelial cells with the receptor-independent stimulus thimerosal (5 mumol/l, 40 min) evoked a long-lasting release of EDRF and small elevations of [Ca2+]i (twofold) after washout of the drug. Depolarization with K+ decreased thimerosal-induced EDRF release and [Ca2+]i in a reversible manner. In patch-clamped endothelial cells, bradykinin (1 nmol/l) induced transient hyperpolarizations that were significantly prolonged by BRL 34915 (1 mumol/l), an activator of K+ channels. BRL 34915 also elicited increases in [Ca2+]i, particularly in thimerosal-stimulated endothelial cells. These effects were abolished by K+. We conclude that the initial rise in [Ca2+]i in response to receptor-binding agonists, caused by mobilization of Ca2+ from intracellular stores, activates K+ channels, thereby inducing hyperpolarization.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Institut für Angewandte Physiologie, Universität Freiburg, Federal Republic of Germany.
authors.forename: A
authors.initials: A
authors.lastname: Lückhoff
authors.forename: R
authors.initials: R
authors.lastname: Busse
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology; D001920:Bradykinin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism / Q000493:pharmacokinetics*; D002417:Cattle; D002463:Cell Membrane Permeability / Q000502:physiology; D002478:Cells, Cultured; D019806:Cromakalim; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D006706:Homeostasis / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism*; D010276:Parasympatholytics / Q000494:pharmacology; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology; D011758:Pyrroles / Q000494:pharmacology; D013849:Thimerosal / Q000494:pharmacology
chemical_list: D001578:Benzopyrans; D010276:Parasympatholytics; D015221:Potassium Channels; D011758:Pyrroles; D019806:Cromakalim; D013849:Thimerosal; D009569:Nitric Oxide; D000255:Adenosine Triphosphate; D000109:Acetylcholine; D001920:Bradykinin; D002118:Calcium