Coupling of excitation to Ca²⁺release is modulated by dysferlin

Key points Dysferlin, the protein missing in limb girdle muscular dystrophy 2B and Miyoshi myopathy, concentrates in transverse tubules of skeletal muscle, where it stabilizes voltage‐induced Ca 2+ transients against loss after osmotic shock injury (OSI). Local expression of dysferlin in dysferlin‐null myofibres increases transient amplitude to control levels and protects them from loss after OSI. Inhibitors of ryanodine receptors (RyR1) and L‐type Ca 2+ channels protect voltage‐induced Ca 2+ transients from loss; thus both proteins play a role in injury in dysferlin's absence. Effects of Ca 2+ ‐free medium and S107, which inhibits SR Ca 2+ leak, suggest the SR as the primary source of Ca 2+ responsible for the loss of the Ca 2+ transient upon injury. Ca 2+ waves were induced by OSI and suppressed by exogenous dysferlin. We conclude that dysferlin prevents injury‐induced SR Ca 2+ leak. Abstract Dysferlin concentrates in the transverse tubules of skeletal muscle and stabilizes Ca 2+ transients when muscle fibres are subjected to osmotic shock injury (OSI). We show here that voltage‐induced Ca 2+ transients elicited in dysferlin‐null A/J myofibres were smaller than control A/WySnJ fibres. Regional expression of Venus‐dysferlin chimeras in A/J fibres restored the full amplitude of the Ca 2+ transients and protected against OSI. We also show that drugs that target ryanodine receptors (RyR1: dantrolene, tetracaine, S107) and L‐type Ca 2+ channels (LTCCs: nifedipine, verapamil, diltiazem) prevented the decrease in Ca 2+ transients in A/J fibres following OSI. Diltiazem specifically increased transients by ∼20% in uninjured A/J fibres, restoring them to control values. The fact that both RyR1s and LTCCs were involved in OSI‐induced damage suggests that damage is mediated by increased Ca 2+ leak from the sarcoplasmic reticulum (SR) through the RyR1. Congruent with this, injured A/J fibres produced Ca 2+ sparks and Ca 2+ waves. S107 (a stabilizer of RyR1–FK506 binding protein coupling that reduces Ca 2+ leak) or local expression of Venus‐dysferlin prevented OSI‐induced Ca 2+ waves. Our data suggest that dysferlin modulates SR Ca 2+ release in skeletal muscle, and that in its absence OSI causes increased RyR1‐mediated Ca 2+ leak from the SR into the cytoplasm.