STED analysis reveals the organization of nonmuscle muscle II, muscle myosin II, and F-actin in nascent myofibrils.

A three-step model has been proposed to describe myofibril assembly in vertebrate cardiac and skeletal muscle cells beginning with premyofibrils, followed by nascent myofibrils, and ending as mature myofibrils (reviewed in Sanger, Wang, et al. (2017). Assembly and maintenance of myofibrils in striated muscle. Handbook of Experimental Pharmacology 235, 39-75; Wang, Fan, (2020). Myofibril assembly and the roles of the ubiquitin proteasome system. Cytoskeleton 77, 456-479). Premyofibrils are composed of minisarcomeres that contain nonmuscle myosin II filaments interdigitating with actin filaments embedded at their barbed ends in muscle-specific alpha-actinin-rich Z-bodies. Sarcomeres in mature myofibrils have filaments of muscle myosin II that interact with actin filaments that are attached to muscle alpha-actinin in Z-bands. Nascent myofibrils, the transitional step between premyofibrils and mature myofibrils, possess two types of myosins II, that is, nonmuscle myosin II and muscle myosin II. The relationship of these two different myosins II in nascent myofibrils, however, is not clear. Stimulated emission depletion (STED) microscopic analyses of nascent myofibrils in both embryonic chick cardiomyocytes, and hiPSC-derived cardiomyocytes revealed that nonmuscle myosin II is in the middle of the nascent myofibril, surrounded by overlapping muscle myosin II filaments at the periphery, and non-striated filamentous actin is present in the nascent myofibril. These findings support the original three-step model of myofibril assembly proposed by Rhee, Sanger, and Sanger, (1994). The premyofibrils: Evidence for its role in myofibrillogenesis. Cell Motility and the Cytoskeleton 28, 1-24.