Formation of oriented myofibrils is a key event in musculoskeletal development. However, the mechanisms that drive myocyte orientation and fusion to control muscle directionality in adults remain enigmatic. Here, we demonstrate that the developing skeleton instructs the directional outgrowth of skeletal muscle and other soft tissues during limb and facial morphogenesis in zebrafish and mouse. Time-lapse live imaging reveals that during early craniofacial development, myoblasts condense into round clusters corresponding to future muscle groups. These clusters undergo oriented stretch and alignment during embryonic growth. Genetic perturbation of cartilage patterning or size disrupts the directionality and number of myofibrils in vivo. Laser ablation of musculoskeletal attachment points reveals tension imposed by cartilage expansion on the forming myofibers. Application of continuous tension using artificial attachment points, or stretchable membrane substrates, is sufficient to drive polarization of myocyte populations in vitro. Overall, this work outlines a biomechanical guidance mechanism that is potentially useful for engineering functional skeletal muscle.

Center for iPS Cell Research and Application Kyoto University Kyoto 606 8507 Japan

Central European Institute of Technology Brno University of Technology Brno Czech Republic

Department of Medical Biochemistry and Biophysics Division of Molecular Neurobiology Karolinska Institutet 17177 Stockholm Sweden

Department of Molecular Biosciences Wenner Gren Institute Stockholm University 10691 Stockholm Sweden

Department of Neuroimmunology Center for Brain Research Medical University Vienna 1090 Vienna Austria

Department of Neuroscience Karolinska Institutet 17177 Stockholm Sweden

Department of Physics University of Erlangen Nuremberg 91052 Erlangen Germany

Department of Physiology and Pharmacology Karolinska Institutet 17177 Stockholm Sweden

Institute for the Advanced Study of Human Biology Kyoto University Kyoto 606 8501 Japan

KTH Royal Institute of Technology SE 100 44 Stockholm Sweden

Laboratory of Biomechanics Institute for Life and Medical Sciences Kyoto University Kyoto 606 8507 Japan

Max Planck Institute for Evolutionary Biology August Thienemann Str 2 24306 Plön Germany

Spinalis Foundation 169 70 Solna Sweden

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