Rapid Calcium Release from the Isolated Sarcoplasmic Reticulum is Triggered Via the Attached Transverse Tubular System

Rapid replacement of 0.15 M K gluconate with 0.15 M choline Cl led to multiphasic Ca2+ release from a heavy fraction of rabbit skeletal muscle microsomes. Following the initial lag period (0-50 ms), about 15 nmol of Ca2+/mg of protein was rapidly released with first-order rate constants k = 60-140 s-1. Subsequently, a larger amount of Ca2+ (up to 56 nmol/mg) was released at a slower rate (k = 0.8-1.5 s-1). The Ca2+ released in both rapid and slow phases was reaccumulated within 60 s. In agreement with a previous report (Caswell, A. H., Lau, Y. H., Garcia, M., and Brunschwig, J-P. (1979) J. Biol. Chem. 254, 202-208), French press treatment of the tubule/sarcoplasmic reticulum (SR) complex results in dissociation of transverse tubular membrane (T-tubules) from SR. Subsequent incubation with 0.4 M potassium cacodylate results in the reassociation of the complex, as shown by sucrose density-gradient sedimentation. Upon T-tubule dissociation, both rapid and slow Ca2+ release was inhibited. Upon reassociation, the rapid Ca2+ release was completely restored and the slow phase partially restored. The results indicate that the T-tubule associated with SR plays a crucial role in triggering rapid Ca2+ release induced by ionic replacement. Other types of Ca2+ release, e.g. those induced by Ca2+ alone or with drugs such as caffeine and quercetin, are unaffected by T-tubule dissociation, and hence produced by direct stimulation of the SR membrane.