Spontaneous calcium release from sarcoplasmic reticulum. Assessment of other ionic influences.

Abstract

A form of spontaneous Ca2+ release from purified light sarcoplasmic reticulum has been recently described (Palade, P., Mitchell, R. D., and Fleischer, S. (1983) J. Biol. Chem. 258, 8098-8107). It is characterized by rapid Ca2+ efflux (1-10 mumol/min X mg protein) which begins only after a delay following preloading and depletion of extravesicular Ca2+. In the present study, the influences of a number of ionic factors modulating the spontaneous calcium release phenomenon are described. The divalent metal ions Mn2+ and Sr2+ and higher external [Mg2+] inhibit release. Mn2+ appears to inhibit from outside, whereas Sr2+ must be taken up to inhibit. Decreasing the phosphate concentration during preloading eliminates spontaneous release, but it can be partially restored with salts of other nonprecipitating anions. No such release was obtained with other Ca2+-precipitating anions, i.e. pyrophosphate, oxalate, and fluoride. The release characteristics are strongly dependent on the monovalent cation present. Spontaneous Ca2+ release is optimal at pH 6.8-7.0 and decreases sharply at higher and lower pH. Ca2+ release can be enhanced by addition, during the lag period, of concentrated salt solutions consisting of more permeant cations than anions, which may indicate that an inside positive membrane potential enhances the rate of calcium release. Spontaneous Ca2+ release can take place in the presence of sucrose, and Ca2+ efflux is not accompanied by a corresponding efflux of preloaded sucrose, demonstrating that the vesicles remain sealed during spontaneous release. The spontaneous Ca2+ release process is distinct from several other forms of Ca2+ release from sarcoplasmic reticulum.