Chemical Depolarization-Induced SR Calcium Release in Triads Isolated from Rabbit Skeletal Muscle

• Published in: Biochemistry (Easton) Vol. 33; no. 36; pp. 10961 - 10968
• Main Authors: Ikemoto, Noriaki, Yano, Masafumi, El-Hayek, Roque, Antoniu, Bozena, Morii, Magotoshi
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 13-09-1994
• Subjects: Animals; Biological Transport; Calcium - metabolism; In Vitro Techniques; Ions; Membrane Potentials; Microsomes - metabolism; Rabbits; Sarcoplasmic Reticulum - metabolism; Solutions; Space life sciences
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00202a015

Excitation-Ca2+ release coupling properties in the heavy microsomal fraction of the rabbit skeletal muscle enriched in triads were investigated by following the same type of approach used for the studies of excitation-contraction coupling in the skinned fiber system. Incubation of the triads with Mg-ATP in a solution containing 150 mM K+, 15.0-37.2 mM Na+, 150-180 mM gluconate-, and 150-200 microM Ca2+ (priming solution) led to (a) the generation of a T-tubule membrane potential making the cytoplasmic side negative, as assessed by potential-dependent uptake of the potential probe [14C]SCN- by triads, and (b) active transport of Ca2+ into the SR moiety. One volume of the primed (viz., polarized and Ca(2+)-loaded) triads was mixed with nine volumes of depolarization solution according to Cl(-)-replacement [Donaldson, S.K.B. (1985) J. Gen. Physiol 86, 501-525; Stephenson, E. W. (1985) J. Gen. Physiol. 86, 813-832] and Na(4)-replacement [Lamb, G.D., & Stephenson, D.G. (1990) J. Physiol. 423, 495-517] protocols used for the induction of contraction in skinned fiber system. The ionic replacement procedure by either protocol produced a rapid release of Ca2+ from SR as determined by stopped-flow fluorometry using fluo-3 as a Ca2+ probe in the presence of BAPTA-calcium buffer. Both the rate constant and the magnitude of Ca2+ release increased with the degree of ionic replacement. The ionic replacement-dependent changes in the release kinetics showed a striking similarity to the voltage-dependent changes of the Ca2+ transient in the intact fiber system.