To analyze vasopressin-induced Ca2+ increase in liver cells, rat hepatocytes were isolated and attached to collagen-coated cover slips. Using fura-2, a Ca2+-sensing dye, changes in intracellular Ca2+ concentration by vasopressin were monitored. Results in this communication suggested that vasopressin-induced Ca2+ increase were composed of both Ca2+ release from internal Ca2+ stores and influx from the plasma membrane. The Ca2+ influx consisted of two distinguishable components. One was dependent on the presence of vasopressin and the other was not. SK&F96365 blocked vasopressin-induced Ca2+ influx in a dose-dependent manner. Vasopressin-induced Ca2+ release from internal stores diminished in a primary culture of hepatocytes according to the culture time. However, changes in vasopressin-induced Ca2+ influx across the plasma membrane differed from changes in the Ca2+ release from internal stores, suggesting two separate signalings from receptor activation to internal stores and to the plasma membrane.