Analyzing the role of dysferlin in skeletal muscle in membrane repair
Our research is focuses on the role of dysferlin in resealing of the skeletal muscle sarcolemmal membrane after that membrane is disrupted. The studies in this project utilize dysferlin variants encoded by cDNAs carrying different point mutations or missing specific exonic sequences or structural domains, many of which are based on variants seen in dysferlinopathy patients. Expression constructs for these dysferlin variants were delivered into flexor digitorum brevis (FDB) myofibers in mice by electroporation and then assayed for their ability to reseal their sarcolemmal membranes by a laser membrane wounding assay. Several laboratories use a similar laser wounding assay to study the role of dysferlin and membrane repair in skeletal muscle. Analysis of the function of these various dysferlin variants in membrane resealing shows that specific variant response differently during the membrane repair response. These results and those planned will help establish the extent that changes in membrane repair capacity contribute to the progression of dysferlinopathy. Understanding the contribution of compromised membrane repair to dysferlinopathies should allow for more effective design of therapeutic approaches to treat these muscular dystrophies.