NA+-H+ ANTIPORT DETECTED THROUGH HYDROGEN-ION CURRENTS IN RAT ALVEOLAR EPITHELIAL-CELLS AND HUMAN NEUTROPHILS

被引:35
作者
DECOURSEY, TE [1 ]
CHERNY, VV [1 ]
机构
[1] RUSSIAN ACAD SCI, AN FRUMKIN ELECTROCHEM INST, MOSCOW, RUSSIA
关键词
D O I
10.1085/jgp.103.5.755
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Voltage-activated H+-selective currents were studied in cultured adult rat alveolar epithelial cells and in human neutrophils using the whole-cell configuration of the patch-clamp technique. The H+ conductance, g(H), although highly selective for protons, was modulated by monovalent cations. In Na+ and to a smaller extent in Li+ solutions, H+ currents were depressed substantially and the voltage dependence of activation of the g(H) shifted to more positive potentials, when compared with the ''inert'' cation tetramethylammonium (TMA(+)). The reversal potential of the g(H), V-rev, was more positive in Na+ solutions than in inert ion solutions. Amiloride at 100 mu M inhibited H+ currents in the presence of all cations studied except Li+ and Na+, in which it increased H+ currents and shifted their voltage-dependence and V-rev, to more negative potentials. The more specific Na+-H+ exchange inhibitor dimethylamiloride (DMA) at 10 mu M similarly reversed most of the suppression of the g(H) by Na+ and Li+. Neither 500 mu M amiloride nor 200 mu M DMA added internally via the pipette solution were effective. Distinct inhibition of the g(H) was observed with 1% [Na+](o), indicating a mechanism with high sensitivity. Finally, the effects of Na+ and their reversal by amiloride were large when the proton gradient was outward (pH(o) parallel to pH(i) 7 parallel to 5.5), smaller when the proton gradient was abolished (pH 7 parallel to 7), and absent when the proton gradient was inward (pH 6 parallel to 7). We propose that the effects of Na+ and Li+ are due to their transport by the Na+-H+ antiporter, which is present in both cell types studied. Electrically silent H+ efflux through the antiporter would increase pH(i) and possibly decrease local pH(o), both of which modulate the g(H) in a similar manner: reducing the H+ currents at a given potential and shifting their voltage-dependence to more positive potentials. A simple diffusion model suggests that Na+-H+ antiport could deplete intracellular protonated buffer to the extent observed. Evidently the Na+-H+ antiporter functions in perfused cells, and its operation results in pH changes which can be detected using the g(H) as a physiological sensor. Thus, the properties of the g(H) can be exploited to study Na+-H+ antiport in single cells under controlled conditions.
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页码:755 / 785
页数:31
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