Platelets are used as models for vascular smooth muscle cells (VSMC) in evaluating intracellular calcium ([Ca2+]i) metabolism in humans. This study was designed to determine if agonist-induced increases in [Ca2+]i in platelets occur via release from intracellular stores as previously demonstrated for VSMC. Incubation of purified platelets loaded with fura-2-AM in media containing 1.5 mmol/L Ca2+ resulted in higher basal [Ca2+]i than platelets incubated in Ca(2+)-free media. In addition, vasopressin-induced platelet [Ca2+]i transients were almost completely blocked by Ca2+ channel blockers. Thus, in contrast to VSMC, the transmembranous flux of extracellular Ca2+ is the major mechanism in vasopressin-induced increases in platelet [Ca2+]i, while mobilization of intracellular Ca2+ stores is only minimally involved.