A focal adhesion protein-based mechanochemical checkpoint regulates cleft progression during branching morphogenesis.
Cleft formation is the initial step of branching morphogenesis in many organs. We previously demonstrated that ROCK 1 regulates a nonmuscle myosin II-dependent mechanochemical checkpoint to transition initiated clefts to progressing clefts in developing submandibular salivary glands. Here, we report that ROCK-mediated integrin activation and subsequent formation of focal adhesion complexes comprise this mechanochemical checkpoint. Inhibition of ROCK1 and nonmuscle myosin II activity decreased integrin β1 activation in the cleft region and interfered with localization and activation of focal adhesion complex proteins, such as focal adhesion kinase (FAK). Inhibition of FAK activity also prevented cleft progression, by disrupting recruitment of the focal adhesion proteins talin and vinculin and subsequent fibronectin assembly in the cleft region while decreasing ERK1/2 activation. These results demonstrate that inside-out integrin signaling leading to a localized recruitment of active FAK-containing focal adhesion protein complexes generates a mechanochemical checkpoint that facilitates progression of branching morphogenesis.
Graduate program in Molecular, Cellular, Developmental, and Neural Biology, University at Albany, State University of New York, Albany, New York, USA.
William P
WP
Daley
Joshua M
JM
Kohn
Melinda
M
Larsen
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005353:Fibronectins / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D022001:Focal Adhesions / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D024462:Myosin Type II / Q000378:metabolism; D009924:Organ Culture Techniques; D011247:Pregnancy; D013363:Submandibular Gland / Q000378:metabolism; D016608:Talin / Q000378:metabolism; D016596:Vinculin / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism
D005353:Fibronectins; D019012:Integrin beta1; D016608:Talin; D016596:Vinculin; D051416:Focal Adhesion Protein-Tyrosine Kinases; C515854:Rock1 protein, mouse; D054460:rho-Associated Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D024462:Myosin Type II