We examined whether H(+)-ATPase is involved in the control of cellular action of arginine vasopressin (AVP) to produce adenosine 3', 5'-monophosphate (cAMP) and mobilize cellular free calcium ([Ca2+]i) in rat renal papillary collecting tubule cells in culture. N-Ethylmaleimide (NEM), an inhibitor of H(+)-ATPase, reduced the cellular pH (pHi) dose-dependently. AVP increased cellular cAMP production in a dose-dependent manner. 5 x 10(-6) and 1 x 10(-5) M NEM significantly diminished the AVP-induced increase in cAMP production. 1 x 10(-7) M AVP also increased [Ca2+]i from 111.2 to 189.3 nM, which was significantly reduced by NEM in a dose-dependent manner. These results indicate that H(+)-ATPase participates the cellular action of AVP mediated via the pHi control in renal papillary collecting tubule cells.