The formation of a multinucleated muscle fiber from individual myoblasts is a complex morphological event that requires dramatic cytoskeletal rearrangements. This multistep process includes myoblast fusion, myotube migration and elongation, myotube target recognition, and finally attachment to form a stable adhesion complex. Many of the studies directed towards understanding the developmental process of muscle morphogenesis at the cellular level have relied on forward genetic screens in model systems such as Drosophila melanogaster for mutations affecting individual stages in myogenesis. Through the analyses of these gene products, proteins that regulate the actin or microtubule cytoskeleton have emerged as important players in each of these steps. We recently demonstrated that RacGAP50C, an essential protein that functions as a cytoskeletal regulator during cell division, also plays an important role in organizing the polarized microtubule network in the elongating myotube. Here we review the current literature regarding Drosophila myogenesis and illustrate several steps of muscle development with respect to the diverse roles that the cytoskeleton plays during this process. Furthermore, we discuss the significance of cytoskeletal coordination during these multiple steps.

The microtubule (MT) cytoskeleton is reorganized during myogenesis as individual myoblasts fuse into multinucleated myotubes. Although this reorganization has long been observed in cell culture, these findings have not been validated during development, and proteins that regulate this process are largely unknown. We have identified a novel postmitotic function for the cytokinesis proteins RacGAP50C (Tumbleweed) and Pavarotti as essential regulators of MT organization during Drosophila myogenesis. We show that the localization of the MT nucleator γ-tubulin changes from diffuse cytoplasmic staining in mononucleated myoblasts to discrete cytoplasmic puncta at the nuclear periphery in multinucleated myoblasts, and that this change in localization depends on RacGAP50C. RacGAP50C and γ-tubulin colocalize at perinuclear sites in myotubes, and in RacGAP50C mutants γ-tubulin remains dispersed throughout the cytoplasm. Furthermore, we show that the mislocalization of RacGAP50C in pavarotti mutants is sufficient to redistribute γ-tubulin to the muscle fiber ends. Finally, myotubes in RacGAP50C mutants have MTs with non-uniform polarity, resulting in multiple guidance errors. Taken together, these findings provide strong evidence that the reorganization of the MT network that has been observed in vitro plays an important role in myotube extension and muscle patterning in vivo, and also identify two molecules crucial for this process.

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