The vasorelaxant effect of atrial natriuretic peptide (ANP) in two species of frogs, Rana catesbeiana and R. tigrina, were studied in vitro. Isolated arterial strips were prepared from the aortic loop, dorsal aorta, iliac, and femoral arteries in the frog. These were stimulated to contract with KCl, norepinephrine, or arginine vasotocin. When maximal contraction was attained, ANP was added to assess if it could relax the strips. Data show that ANP is vasorelaxant in these preparations. The sensitivity of these tissues depends on the contractile agents, e.g., KCl-stimulated preparations from R. catesbeiana did not respond to ANP. Data also suggest that ANP action elicited in the frog vascular tissue is due to an inhibitory effect on the mobilization of the intracellular calcium store and/or calcium influx from extracellular space to initiate contraction.