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

Am J Physiol. 1999 Sep;277(3):C469-79. doi: 10.1152/ajpcell.1999.277.3.C469.

Abstract

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 I(Na) and the apical Na+ concentration. The i and N) 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ammonium Chloride / pharmacology
  • Biological Transport / physiology
  • Cell Line
  • Cell Membrane / metabolism
  • Cytosol / metabolism
  • Epithelial Sodium Channels
  • Hydrogen / metabolism*
  • Hydrogen-Ion Concentration
  • Intracellular Membranes / metabolism*
  • Kidney / cytology
  • Kidney / metabolism*
  • Kinetics
  • Sodium / metabolism*
  • Sodium Channel Blockers
  • Sodium Channels / metabolism
  • Sodium Channels / physiology
  • Sodium Chloride / pharmacology
  • Sodium-Hydrogen Exchangers / antagonists & inhibitors

Substances

  • Epithelial Sodium Channels
  • Sodium Channel Blockers
  • Sodium Channels
  • Sodium-Hydrogen Exchangers
  • Ammonium Chloride
  • Sodium Chloride
  • Hydrogen
  • Sodium