A voltage-dependent and pH-sensitive proton current in Rana esculenta oocytes

Voltage clamp technique was used to study macroscopic ionic currents in Rana esculenta oocytes. Depolarization steps led to the activation of a single type of outward current (Iout) when contaminant potassium and calcium-dependent chloride currents were pharmacologically inhibited. The voltage threshold of Iout activation was 10 mV and this current, which did not inactivate, presented a deactivation the time constant of 73 +/- 21 msec (n = 26) corresponding to a membrane voltage of -60 mV. Its reversal potential (Erev) was dependent on the magnitude of the depolarization and also on pulse duration. These changes in Erev were thought to reflect intracellular ion depletion occurring during activation of the remaining outward current. Furthermore, the activation threshold of Iout was clearly affected by modifications in extracellular and intracellular H+ concentrations. Indeed, intracellular alkalinization (evoked by external application of ammonium chloride) or extracellular acidification induced a rightward shift in the activation threshold while intracellular acidification (evoked by external application of sodium acetate) or extracellular alkalinization shifted this threshold toward a more negative value. Lastly, Iout was dramatically reduced by divalent cations such as Cd2+, Ni2+ or Zn2+ and was strongly decreased by 4 Aminopyridine (4-AP), well-known H+ current antagonists already described in many cell types. Therefore, it was suggested that the outward current was prominently carried by H+ ions, which may play a key role in the regulation of intracellular pH and subsequent pH dependent processes in Rana oocyte.