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

Abstract

We present results from a novel study of Ca2+ waves in vascular smooth muscle cells using a sarcoplasmic reticulum (SR)-targeted Ca2+ indicator that specifically binds to the luminal protein calsequestrin.Agonist-stimulated waves of elevated cytoplasmic calcium concentration regulate blood vessel tone, and vasomotion in vascular smooth muscle. Previous studies employing cytoplasmic calcium indicators revealed that these calcium waves are generated by a combination of inositol 1,4,5-trisphosphate (IP3) and calcium-induced calcium release (CICR) from the SR. Our findings confirm that these waves are due to regenerative CICR by the receptors for IP3 (IP3R).The main new finding from our bservations is a transient elevation in luminal SR Ca2+ concentration ([Ca2+]_SR) both at the site of wave initiation, just before regenerative Ca2+ release commences, and at the advancing wave front, during wave propagation. This strongly suggests a role of [Ca2+]_SR in activation of IP3R.In addition, we find that these depletion waves are characterized by a decreasing velocity as they progress. We developed a quantitative diffusional model to analyze this finding and conclude that the gradual decrease in the velocity of the SR Ca2+ depletion wave, observed in the absence of external calcium, indicates continuity of the lumen of the sarcoplasmicreticulum network.Finally, our observation that the depletion wave was arrested by the nuclear envelope may have implications for selective Ca2+ signalling.