Membranes were isolated by isotonic homogenization and differential centrifugation from rat hepatocytes cultured overnight. The specific GTPase activity of the membranes was 1-1.3 pmol gamma-labelled GTP hydrolysed/mg protein per min in the presence of 1.2 mM Na+, 2 mM EGTA, 1 mM ATP and 0.2 mM 5-adenylyl imidodiphosphate. Under these conditions there was a stimulation of specific GTPase activity of no more than 20% by 11-115 nM vasopressin. No effect of vasopressin was seen in the presence of 1.7 microM free Ca2+ or 100 mM Na+. The findings indicate that vasopressin is able to influence GTPase activity as well as accelerate phosphoinositide breakdown in rat hepatocytes.