Coupling of excitation to Ca2+ release is modulated by dysferlin

Abstract

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 Ca²⁺ 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 Ca²⁺ channels protect voltage-induced Ca²⁺ transients from loss; thus both proteins play a role in injury in dysferlin's absence. Effects of Ca²⁺ -free medium and S107, which inhibits SR Ca²⁺ leak, suggest the SR as the primary source of Ca²⁺ responsible for the loss of the Ca²⁺ transient upon injury. Ca²⁺ waves were induced by OSI and suppressed by exogenous dysferlin. We conclude that dysferlin prevents injury-induced SR Ca²⁺ leak.

Abstract: Dysferlin concentrates in the transverse tubules of skeletal muscle and stabilizes Ca²⁺ transients when muscle fibres are subjected to osmotic shock injury (OSI). We show here that voltage-induced Ca²⁺ 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 Ca²⁺ transients and protected against OSI. We also show that drugs that target ryanodine receptors (RyR1: dantrolene, tetracaine, S107) and L-type Ca²⁺ channels (LTCCs: nifedipine, verapamil, diltiazem) prevented the decrease in Ca²⁺ 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 Ca²⁺ leak from the sarcoplasmic reticulum (SR) through the RyR1. Congruent with this, injured A/J fibres produced Ca²⁺ sparks and Ca²⁺ waves. S107 (a stabilizer of RyR1-FK506 binding protein coupling that reduces Ca²⁺ leak) or local expression of Venus-dysferlin prevented OSI-induced Ca²⁺ waves. Our data suggest that dysferlin modulates SR Ca²⁺ release in skeletal muscle, and that in its absence OSI causes increased RyR1-mediated Ca²⁺ leak from the SR into the cytoplasm.

Keywords: dysferlin; excitation-contraction coupling; skeletal muscle.