[Characterization of calcium permeability at steady-state calcium load in masseter muscle sarcoplasmic reticulum].

Abstract

To characterize the route of calcium permeability, the effect of intravesicular and extravesicular calcium concentration on the permeability from sarcoplasmic reticulum (SR) vesicles isolated from canine masseter muscle was determined by measuring net efflux of calcium after stopping pump-mediated fluxes. The apparent permeability, calculated as the net efflux divided by the total intravesicular calcium, depended on calcium load. When the intravesicular bound calcium was taken into account, net calcium efflux was found to be linearly related to the difference in calcium concentration across the SR membrane. The first order rate constant of calcium permeability was nearly identical when efflux was initiated by the addition of EGTA or glucose plus hexokinase to quench calcium pump by lowering activator calcium or by converting substrate ATP to ADP and glucose 6-phosphate, respectively. Extravesicular calcium concentration between 0.001 microM and 33.9 microM had no great effect on calcium permeability. The results suggest that some minimal calcium gradient may be required in order to observe a substantial passive calcium efflux, and the passive calcium efflux is not carrier mediated. It is also postulated that passive route of efflux during calcium accumulation is relatively small and that physiological calcium release during excitation-contraction coupling does not occur through this route.