Disruption of action potential and calcium signaling properties in malformed myofibers from dystrophin‐deficient mice

Duchenne muscular dystrophy (DMD), the most common and severe muscular dystrophy, is caused by the absence of dystrophin. Muscle weakness and fragility (i.e., increased susceptibility to damage) are presumably due to structural instability of the myofiber cytoskeleton, but recent studies suggest that the increased presence of malformed/branched myofibers in dystrophic muscle may also play a role. We have previously studied myofiber morphology in healthy wild‐type (WT) and dystrophic (MDX) skeletal muscle. Here, we examined myofiber excitability using high‐speed confocal microscopy and the voltage‐sensitive indicator di‐8‐butyl‐amino‐naphthyl‐ethylene‐pyridinium‐propyl‐sulfonate (di‐8‐ANEPPS) to assess the action potential (AP) properties. We also examined AP‐induced Ca2+ transients using high‐speed confocal microscopy with rhod‐2, and assessed sarcolemma fragility using elastimetry. AP recordings showed an increased width and time to peak in malformed MDX myofibers compared to normal myofibers from both WT and MDX, but no significant change in AP amplitude. Malformed MDX myofibers also exhibited reduced AP‐induced Ca2+ transients, with a further Ca2+ transient reduction in the branches of malformed MDX myofibers. Mechanical studies indicated an increased sarcolemma deformability and instability in malformed MDX myofibers. The data suggest that malformed myofibers are functionally different from myofibers with normal morphology. The differences seen in AP properties and Ca2+ signals suggest changes in excitability and remodeling of the global Ca2+ signal, both of which could underlie reported weakness in dystrophic muscle. The biomechanical changes in the sarcolemma support the notion that malformed myofibers are more susceptible to damage. The high prevalence of malformed myofibers in dystrophic muscle may contribute to the progressive strength loss and fragility seen in dystrophic muscles. In Duchenne muscular dystrophy (DMD), muscle weakness and fragility are presumably due to structural instability of the myofiber cytoskeleton, but recent studies suggest that the increased presence of malformed/branched myofibers in dystrophic muscle may also play a role. This study, using the murine model for DMD, supports the notion that malformed myofibers are functionally different from myofibers with normal morphology. The differences seen in action potential properties, calcium signals, and membrane biomechanics indicate changes in excitability, remodeling of the