Mechanisms of reduced SR Ca²⁺ release induced by inorganic phosphate in rat skeletal muscle fibers

Abstract

The effects of inorganic phosphate (P(i)) on Ca(2+) release from the sarcoplasmic reticulum (SR) were studied in mechanically skinned rat skeletal muscle fibers. Application of caffeine or T-tubule depolarization was used to induce Ca(2+) release from the SR, which was detected using fura 2 fluorescence. Addition of P(i) (1-40 mM) caused a reversible and concentration-dependent reduction in the caffeine-induced Ca(2+) transient. This effect was apparent at low P(i) concentration (<5 mM), which did not result in detectable precipitation of calcium phosphate within the SR. The inhibitory effect of P(i) exhibited a marked dependence on free Mg(2+) concentration. At 0.5 mM free Mg(2+), 5 mM P(i) reduced the caffeine-induced transient by 25.1 +/- 4.1% (n = 13). However, at 1.5 mM free Mg(2+), 5 mM P(i) reduced the amplitude of caffeine-induced Ca(2+) transients by 68.9 +/- 3.1% (n = 10). Depolarization-induced SR Ca(2+) release was similarly affected. These effects of P(i) may be important in skeletal muscle fatigue, if an inhibitory action of P(i) on SR Ca(2+) release is augmented by the rise in cytosolic Mg(2+) concentration, which accompanies ATP breakdown.