Treatment with the calcium ionophore A23187 on either the serosal or mucosal sides of frog skin, strongly inhibits the hydrosmotic response to vasopressin. On the contrary, the hydrosmotic response to 8-br-cAMP is not affected by treatment with the A23187. Trifluoperazine, a drug which inhibits the Ca2+-calmodulin complex, selectively inhibits vasopressin-induced water transport. Collectively, our results suggest that an increase in the intracellular concentration of Ca2+, obtained by treatment with the ionophore A23187, interferes with a pre-cAMP step of the hydrosmotic response to the antidiuretic hormone. Calcium ions could regulate adenyl-cyclase activity and consequently intracellular levels of cAMP. This effect may probably involve calmodulin.