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Am J Physiol Cell Physiol 277: C469-C479, 1999;
0363-6143/99 $5.00
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Vol. 277, Issue 3, C469-C479, September 1999

Intracellular pH shifts in cultured kidney (A6) cells: effects on apical Na+ transport

Wolfgang Zeiske1, Ilse Smets2, Marcel Ameloot2, Paul Steels2, and Willy van Driessche1

2 Laboratory of Physiology, Limburgs Universitair Centrum, B-3590 Diepenbeek; and 1 Laboratory of Physiology, Katholieke Universiteit Leuven, Campus Gasthuisberg, B-3000 Leuven, Belgium

We report, for the epithelial Na+ channel (ENaC) in A6 cells, the modulation by cell pH (pHc) of the transepithelial Na+ current (INa), the current through the individual Na+ channel (i), the open Na+ channel density (No), and the kinetic parameters of the relationship between INa and the apical Na+ concentration. The i and No were evaluated from the Lorentzian INa noise induced by the apical Na+ channel blocker 6-chloro-3,5-diaminopyrazine-2-carboxamide. pHc shifts were induced, under strict and volume-controlled experimental conditions, by apical/basolateral NH4Cl pulses or basolateral arrest of the Na+/H+ exchanger (Na+ removal; block by ethylisopropylamiloride) and were measured with the pH-sensitive probe 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein. The changes in pHc were positively correlated to changes in INa and the apically dominated transepithelial conductance. The sole pHc-sensitive parameter underlying INa was No. Only the saturation value of the INa kinetics was subject to changes in pHc. pHc-dependent changes in No may be caused by influencing Po, the ENaC open probability, or/and the total channel number, NT = No/Po.

noise analysis; single-channel current; epithelial sodium channel; ammonium; cell volume


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