Visualization of the subsarcolemmal cytoskeleton network of mouse skeletal muscle cells by en face views and application to immunoelectron localization of dystrophin

The ultrastructural organization of the highly interconnected filamentous network underneath the sarcolemma as well as the role played by the muscle protein dystrophin within this cytoskeleton remain yet unclear. More accurate information has been obtained by using a method which provides three-dimensional en face views of large membrane areas applied to mouse cultured myotubes and isolated adult skeletal muscle fibres. Two levels have been distinguished in the cytoskeleton underlying the sarcolemma: the submembranous level, partly integrated into the membrane, and the cortical level, invading the proximal cytoplasmic space. Few differences have been found between the membrane cytoskeletons of myotubes issued from 14-day-old cultures and those of adult fibres. The comparison was done with cells where dystrophin is missing (mdx mouse muscle): surprisingly, the lack of dystrophin does not induce obvious or dramatic ultrastructural disorganization, either in the cortical cytoskeletal network or in the submembranous one. Immunogold labelling of either the central-rod or the C-terminal domain of dystrophin is not located among the cortical network. This study provides additional data on the spatial ordering of subsarcolemmal cytoskeletal elements: dystrophin does not appear as a filamentous structure entirely located among subsarcolemmal cytoskeleton but seems partly embedded in membranous material.