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Agonist-stimulated smooth muscle Ca 2+  waves regulate blood vessel tone and vasomotion. Previous studies employing cytoplasmic Ca 2+  indicators revealed that these Ca 2+  waves were stimulated by a combination of inositol 1,4,5-trisphosphate- and Ca 2+ -induced Ca 2+  release from the endo/sarcoplasmic reticulum. Herein, we present the first report of endothelin-1 stimulated waves of Ca 2+  depletion from the sarcoplasmic reticulum of vascular smooth muscle cells using a calsequestrin-targeted Ca 2+  indicator. Our findings confirm that these waves are due to regenerative Ca 2+ -induced Ca 2+  release by the receptors for inositol 1,4,5-trisphosphate. Our main new finding is a transient elevation in SR luminal Ca 2+  concentration ([Ca 2+ ] SR ) both at the site of wave initiation, just before regenerative Ca 2+  release commences, and at the advancing wave front, during propagation. This strongly suggests a role for [Ca 2+ ] SR  in the activation of inositol 1,4,5-trisphosphate receptors during agonist-induced calcium waves. In addition, quantitative analysis of the gradual decrease in the velocity of the depletion wave, observed in the absence of external Ca 2+ , indicates continuity of the lumen of the sarcoplasmic reticulum network. Finally, our observation that the depletion wave was arrested by the nuclear envelope may have implications for selective Ca 2+  signalling.

Waves of Calcium Depletion in the Sarcoplasmic Reticulum of Vascular Smooth Muscle Cells: An Inside View of Spatiotemporal Ca2+ Regulation