In rings of rabbit facial vein (RFV), depletion of sarcoplasmic reticulum (SR) Ca2+ by caffeine abolished the subsequent isometric contraction to 25 mM K+ physiological salt solution (25K-PSS). However, the associated steady-state increase of smooth muscle intracellular free Ca2+concentration ([Ca2+]i), measured using fura PE3 and cuvette photometry, was not altered. Treatment with the specific SR Ca2+ pump inhibitor cyclopiazonic acid (30 μM) after caffeine-induced SR Ca2+ depletion restored and greatly augmented the 25K-PSS-induced contraction. This suggests that SR Ca2+ depletion leads to a dissociation of K+-induced [Ca2+]iincrease from contraction that was dependent on Ca2+ pump-mediated SR Ca2+ uptake. Endothelium removal augmented the 25K-PSS-induced [Ca2+]iincrease after caffeine-induced SR Ca2+ depletion. However, this was associated with only a small and transient contraction. Exposure of endothelium-denuded RFV to cyclopiazonic acid after caffeine-induced SR Ca2+ depletion further amplified the 25K-PSS-induced [Ca2+]iincrease, which was associated with a large and sustained contraction. However, the latter [Ca2+]iincrease was still higher than in endothelium-intact RFV. This suggests that the endothelium dampens the [Ca2+]irise associated with K+-induced Ca2+ influx, but independently of Ca2+ pump-mediated SR Ca2+ uptake.