Dysferlin Regulates Cardiac T-tubule Structure and Excitation-contraction Coupling in Isolated Cardiac Myocytes at Rest and in Response to Acute Hypo-osmotic Stress and is Protective Against Arrhythmias in Langendorff-perfused Hearts

Abstract

Dysferlin is a membrane-associated protein that supports skeletal muscle function such that mutations in the DYSF gene can cause muscular dystrophy. Growing evidence suggests dysferlin regulates cardiac function, but this is less well understood. Tight regulation of the cardiac transverse-(T)-tubule network and excitation-contraction (EC) coupling mechanism is essential for healthy cardiac physiology. Remodelling of T-tubules and the EC coupling mechanism is observed during periods of cardiac stress and pathologies, promoting arrhythmias. However, little is known about how these processes are regulated and any protective mechanisms which may limit detrimental effects. Using a global dysferlin knockout (KO) mouse we have shown that the loss of dysferlin leads to a decrease in T-tubule density and the amplitude and rate of decay of the systolic Ca2+ transient but a narrowing of the dyadic cleft. Electrical mapping of ex vivo DYSF KO hearts shows they are more susceptible to ventricular arrhythmias. To induce stress, we used hypo-osmotic shock injury (OSI) to damage T-tubule networks in cardiac myocytes, in vitro. OSI increased T-tubule fragmentation and caused dysregulation of intracellular Ca2+ handling in dysferlin KO cells relative to WT controls. Finally, we observed that a natural decline in WT cardiac dysferlin abundance, which may contribute to the natural age-dependent maladaptive T-tubule remodelling that occurs in the mammalian ventricle. In summary, these findings demonstrate an essential role for dysferlin in cardiac physiology, especially during conditions of stress, which is decreased in normal ageing.