Formation of oriented myofibrils is a key event in development of skeletal muscles. However, the knowledge of mechanisms that control the orientation of myocyte fusion and the resulting directionality of muscle primordia remains incomplete. Here, we utilized in vivo and in vitro live imaging, 2D and 3D muscle cell culture, CAS9/CRISPR-mediated mutagenesis in fish and genetic experiments in mice to demonstrate that the early steps of myocyte polarization are dependent on the physical forces exerted by cartilage expansion.

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