Characteristics of phosphate-induced Ca2+ efflux from the SR in mechanically skinned rat skeletal muscle fibers

The effects of P-i on sarcoplasmic reticulum (SR) Ca2+ regulation were stud ied in mechanically skinned rat skeletal muscle fibers. Brief application o f caffeine was used to assess the SR Ca2+ content, and changes in concentra tion of Ca2+ ([Ca2+]) within the cytosol were detected with fura 2 fluoresc ence. Introduction of P-i (1-40 mM) induced a concentration-dependent Ca2efflux from the SR. In solutions lacking creatine phosphate (CP), the ampli tude of the P-i-induced Ca2+ transient approximately doubled. A similar pot entiation of P-i-induced Ca2+ release occurred after inhibition of creatine kinase (CK) with 2,4-dinitrofluorobenzene. In the presence of ruthenium re d or ryanodine, caffeine-induced Ca2+ release was almost abolished, whereas P-i-induced Ca2+ release was unaffected. However, introduction of the SR C a2+ ATPase inhibitor cyclopiazonic acid effectively abolished P-i-induced C a2+ release. These data suggest that P-i induces Ca2+ release from the SR b y reversal of the SR Ca2+ pump but not via the SR Ca2+ channel under these conditions. If this occurs in intact skeletal muscle during fatigue, activa tion of a Ca2+ efflux pathway by P-i may contribute to the reported decreas e in net Ca2+ uptake and increase in resting [Ca2+].