Journal
JOURNAL OF CELLULAR PHYSIOLOGY
Volume 212, Issue 2, Pages 473-480Publisher
WILEY
DOI: 10.1002/jcp.21043
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- Wellcome Trust Funding Source: Medline
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When examining HEK293 cells by whole-cell patch-clamp electrophysiology we found spontaneous currents, present in almost all cells. These currents were carried by Na+ ions, were inhibited by amiloride and by cells exposure to acidic (pH 6.3) extracellular solutions. These properties (ion carrier, amiloride-sensitivity, and inactivation by constant lowering of extracellular pH) were similar to the properties of proton-activated currents measured from the same cells. Spontaneous currents required intracellular ATP, were completely inhibited by intracellular Ca2+ buffering with BAPTA and were suppressed by intracellular administration of vesicular H-ATPase inhibitor bafilomycin. ATP-incluced Ca2+ influx through P2X receptors in HEK293 cells stably transfected with P2X2, P2X(2/3) or P2X(4) purinoreceptor subunits transiently potentiated amplitude and frequency of spontaneous currents; this effect was antagonized by bafilomycin. We concluded that spontaneous currents represent activation of acid-sensitive ion channels (ASICs) by autocrine vesicular release of protons from HEK cells.
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