Myoblast cell fusion is essential for skeletal muscle development and regeneration. Yet, the molecular machinery that drives myoblast fusion remains incompletely understood. Myoblast cell fusion is an intricate multistep process, making it challenging to identify the specific proteins involved. Until now, no approach was available to capture fusing cells and dissect the dynamic changes in their cellular transitions. To fill this gap, we have developed a method using small-molecule inhibitors to synchronize muscle differentiation ex vivo and capture cells before, during, and after fusion. This allows us to identify and associate proteins with specific stages of muscle cell differentiation and fusion. Using this method, we have identified the Paralemmin A-kinase anchor protein (PALM2-AKAP2), a protein of unknown function, as a potential regulator of muscle regeneration. Hence, this work provides valuable data and will provide new insight into the mechanism of myoblast fusion and muscle regeneration.
