Unrepeatable extracellular Ca2+-dependent contractile effects of cyclopiazonic acid in rat vascular smooth muscle

Cyclopiazonic acid (CPA), a specific reversible inhibitor of Ca(2+)-pumps in sarcoplasmic reticulum, causes a slowly developing and subsequently diminishing characteristic contraction in endothelium-denuded rat vascular smooth muscle. We recently found that CPA-induced contractions were not completely repeatable in endothelium-denuded rat aorta and superior mesenteric artery. 10 microM CPA-induced contractions expressed as a percentage of 80 mM KCl-induced contraction were significantly decreased from 51.4+/-5.7% to 11.8+/-2.6% (P<0.0001) upon the second application in endothelium-denuded rat aorta, and this was not due to any irreversible cytotoxic effects of CPA. The decrease of CPA-induced contractile responses upon the second application was dependent on both types of blood vessels and doses of CPA upon the first application. CPA upon the second application in Ca(2+)-containing solutions did induce its characteristic contractions in the rings pretreated with Ca(2+)-free solutions or Ca(2+) entry blockers before and during its first application, suggesting that capacitative mode of Ca(2+) influx during the application of CPA might be responsible for the diminishment of contractions upon the second application. These data suggest that CPA by inducing a transient rise in cytosolic Ca(2+) level might cause a long-lasting upregulation of Ca(2+) extrusion across the plasma membrane in vascular smooth muscle cells and thus accelerate Ca(2+) efflux over a prolonged period, leading to unrepeatable contractile effects of CPA. Such long-lasting upregulation of Ca(2+) extrusion may contribute to the regulation of excitability of vascular smooth muscle cells and protect the cells against excitotoxic injury.