Skeletal Muscle Injury by Electroporation: A Model to Study Degeneration/Regeneration Pathways in Muscle

Abstract

Skeletal muscle has a remarkable capacity to regenerate after injuries mainly due to a reservoir of precursor cells named satellite cells (SCs), which are responsible for after-birth growth and response to lesions, either by exercise or disease. Upon injury, the regenerative response includes SCs exit of quiescence, activation, proliferation, and fusion to repair or form new myofibers. This process is accompanied by inflammation, with infiltration of immune cells, primarily macrophages. Every phase of regeneration is highly regulated and orchestrated by many molecules and signaling pathways. The elucidation of players and mechanisms involved in muscle degeneration and regeneration is of extreme importance, especially for therapeutic strategies for muscle diseases.

Here we are proposing a model of muscle injury induced by electroporation, which is an efficient method to induce muscle damage in order to follow the steps involved in degeneration and regeneration. Three days after electroporation, the muscle shows prominent signals of degeneration, like areas of necrosis and infiltration of macrophages, followed by regeneration, observed by the presence of centrally nucleated myofibers. After 5 days the regeneration is very active, with small dMyHC positive fibers. Fifteen days later, we observe a general regeneration of the muscle, with fibers with increased diameter after 60 days. This methodology is an easy and simple alternative to induce muscle lesion. It can be employed to study alterations in gene expression and the process of satellite cell recruitment, both in healthy and dystrophic/myopathic animal models for muscular dystrophy.