In Vivo pH Regulation by a Na+/H+ Antiporter in the Halotolerant Alga Dunaliella salina

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Abstract
Na+/H+ exchange activity in whole cells of the halotolerant alga Dunaliella salina can be elicited by intracellular acidification due to addition of weak acids at appropriate external pH. The changes in both intracellular pH and Na+ were followed. Following a mild intracellular acidification, intracellular Na+ content increased dramatically and then decreased. We interpret the phase of Na+ influx as due to the activation of the plasma membrane Na+/H+ antiporter and the phase of Na+ efflux as due to an active Na+ extrusion process. The following observations are in agreement with this interpretation: (a) the Na+ influx phase was sensitive to Li+, which is an inhibitor of the Na+/H+ antiporter, did not require energy, and was insensitive to vanadate; (b) the Na+ efflux phase is energy-dependent and sensitive to the plasma membrane ATPase inhibitor, vanadate. Following intracellular acidification, a drastic decrease in the intracellular ATP content is observed that is reversed when the cells regain their neutral pH value. We suggest that the intracellular acidification-induced change in the internal Na+ concentration is due to a combination of Na+ uptake via the Na+/H+ antiporter and an active, ATPase-dependent, Na+ extrusion. The Na+/H+ antiporter seems, therefore, to play a principal role in internal pH regulation in Dunaliella.