Regulation of intracellular sodium and pH by the electrogenic H+ pump in frog skin.

We have investigated the role of the electrogenic hydrogen ion pump in the regulation of intracellular sodium ion activity (aiNa) and intracellular pH (pHi) in frog skin epithelial cells using double-barreled ion sensitive microelectrodes. When Rana esculenta skin is mounted in an Ussing chamber and bathed in 1 mM Na2SO4 buffered to pH 7.34 with imidazole on the apical side and in normal Ringer on the serosal side, the apical addition of the carbonic anhydrase inhibitor ethoxzolamide (10(-4)M) blocks net H+ ion excretion and Na absorption, producing a depolarization of 25-30 mV of the apical membrane potential (psi mc). We demonstrate that these changes are accompanied by a fall in aiNa from 6.2 +/- 0.5 mmol/l to 3.4 +/- 0.6 mmol/l and an increase in pHi from 7.20 +/- 0.03 to 7.38 +/- 0.08 (n = 12 skins). Voltage clamping psi mc to its control value in the presence of ethoxzolamide restored aiNa but the pHi remained alkaline. Furthermore, the fall in aiNa produced by ethoxzolamide could be mimicked by voltage clamping psi mc towards the value of the Nernst potential for Na at the apical membrane. These results indicate that the maintenance of the cellular Na+ transport pool is dependent on a favourable electrical driving force and counter-current generated by an electrogenic H+ pump at the apical membrane. Addition of amiloride (10(-5) mol/l) or substitution of external Na+ by Mg2+ or K+ caused a hyperpolarization of psi mc and a fall in aiNa.2+ e