Chronic respiratory acidosis stimulates the Vmax of the renal brush border Na-H antiporter. The activation of protein kinase C (PKC) by phorbol esters stimulates the activity of the renal Na-H antiporter. In this study, the hypothesis that PKC plays a role in the adaptive increase of the renal brush border Na-H antiporter activity to respiratory acidosis was tested. In vivo respiratory acidosis was associated with an increase in in vitro Na-H antiporter activity and also with an increase in brush border membrane PKC activity, without changes in PKC activity in cytosol or basolateral membranes. Na-H antiporter activity, assessed as the amiloride-sensitive component of 22Na uptake, was measured in cultured proximal tubule cells exposed to 10% CO2 for 48 h. Na-H antiporter activity was significantly higher in cells exposed to 10% CO2 than in those exposed to 5% CO2. To evaluate the role of PKC, cultured cells were depleted of PKC by exposure to the active phorbol ester phorbol 12-myristate 13-acetate (PMA; 10(-7) or 10(-6) M) for 48 h before exposure to 10% CO2. In the presence of 10% CO2, Na-H antiporter activity was significantly lower in PKC-depleted cells than in control. In addition, sphingosine, an inhibitor of PKC, also prevented the adaptation of the Na-H antiporter to 10% CO2 as compared with 5% CO2. In cells treated with the inactive analog 4 alpha-PMA, 22Na uptake was not different than that in control. PMA-treated cells also had a decrease in Na-H antiporter activity during exposure to 5% CO2.(ABSTRACT TRUNCATED AT 250 WORDS)