title: Unmasking potential intracellular roles for dysferlin through improved immunolabeling methods.
abstract: Mutations in the DYSF gene that severely reduce the levels of the protein dysferlin are implicated in muscle-wasting syndromes known as dysferlinopathies. Although studies of its function in skeletal muscle have focused on its potential role in repairing the plasma membrane, dysferlin has also been found, albeit inconsistently, in the sarcoplasm of muscle fibers. The aim of this article is to study the localization of dysferlin in skeletal muscle through optimized immunolabeling methods. We studied the localization of dysferlin in control rat skeletal muscle using several different methods of tissue collection and subsequent immunolabeling. We then applied our optimized immunolabeling methods on human cadaveric muscle, control and dystrophic human muscle biopsies, and control and dysferlin-deficient mouse muscle. Our data suggest that dysferlin is present in a reticulum of the sarcoplasm, similar but not identical to those containing the dihydropyridine receptors and distinct from the distribution of the sarcolemmal protein dystrophin. Our data illustrate the importance of tissue fixation and antigen unmasking for proper immunolocalization of dysferlin. They suggest that dysferlin has an important function in the internal membrane systems of skeletal muscle, involved in calcium homeostasis and excitation-contraction coupling.
authors: Department of Physiology, University of Maryland School of Medicine, Baltimore, USA.
authors: Joseph A
authors: JA
authors: Roche
authors: Lisa W
authors: LW
authors: Ru
authors: Andrea M
authors: AM
authors: O'Neill
authors: Wendy G
authors: WG
authors: Resneck
authors: Richard M
authors: RM
authors: Lovering
authors: Robert J
authors: RJ
authors: Bloch
mesh_terms: D000818:Animals; D000073939:Dysferlin; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D051379:Mice; D009124:Muscle Proteins / Q000032:analysis / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000648:ultrastructure; D009136:Muscular Dystrophies / Q000175:diagnosis; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: C114839:DYSF protein, human; C497505:Dysf protein, mouse; C547052:Dysf protein, rat; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins