PubMed · 6316556
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F S Southwick, T P Stossel. 1983. Contractile proteins in leukocyte function.. https://pubmed.ncbi.nlm.nih.gov/6316556/
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Myoblast fusion is crucial for the formation, growth, maintenance and regeneration of healthy skeletal muscle. Unfortunately, the molecular machinery, cell behaviors, and membrane and cytoskeletal remodeling events that govern fusion and myofiber formation remain poorly understood. Using time-lapse imaging approaches on mouse C2C12 myoblasts, we identify discrete and specific molecular events at myoblast membranes during fusion and myotube formation. These events include rearrangement of cell shape from fibroblast to spindle-like morphologies, changes in lamellipodial and filopodial extensions during different periods of differentiation, and changes in membrane alignment and organization during fusion. We find that actin-cytoskeleton remodeling is crucial for these events: pharmacological inhibition of F-actin polymerization leads to decreased lamellipodial and filopodial extensions and to reduced myoblast fusion. Additionally, shRNA-mediated inhibition of Nap1, a member of the WAVE actin-remodeling complex, results in accumulations of F-actin structures at the plasma membrane that are concomitant with a decrease in myoblast fusion. Our data highlight distinct and essential roles for actin cytoskeleton remodeling during mammalian myoblast fusion, provide a platform for cellular and molecular dissection of the fusion process, and suggest a functional conservation of Nap1-regulated actin-cytoskeleton remodeling during myoblast fusion between mammals and Drosophila.
The maize mutant lilliputian is characterized by miniature seedling stature, reduced cell elongation, and aberrant root anatomy. Here, we document that root cells of this mutant show several defects in the organization of actin filaments (AFs). Specifically, cells within the meristem lack dense perinuclear AF baskets and fail to redistribute AFs during mitosis. In contrast, mitotic cells of wild-type roots accumulate AFs at plasma membrane-associated domains that face the mitotic spindle poles. Both mitotic and early postmitotic mutant cells fail to assemble transverse arrays of cortical AFs, which are characteristic for wild-type root cells. In addition, early postmitotic cells show aberrant distribution of endoplasmic AF bundles that are normally organized through anchorage sites at cross-walls and nuclear surfaces. In wild-type root apices, these latter AF bundles are organized in the form of symmetrically arranged conical arrays and appear to be essential for the onset of rapid cell elongation. Exposure of wild-type and cv. Alarik maize root apices to the F-actin drugs cytochalasin D and latrunculin B mimics the phenotype of lilliputian root apices. In contrast to AFs, microtubules are more or less normally organized in root cells of lilliputian mutant. Collectively, these data suggest that the LILLIPUTIAN protein, the nature of which is still unknown, impinges on plant development via its action on the actin cytoskeleton.
Patterning of the Xenopus gastrula marginal zone in the axis running equatorially from the Spemann organizer-the so--called "dorsal/ventral axis"--has been extensively studied. It is now evident that patterning in the animal/vegetal axis also needs to be taken into consideration. We have shown that an animal/vegetal pattern is apparent in the marginal zone by midgastrulation in the polarized expression domains of Xenopus brachyury (Xbra) and Xenopus nodal-related factor 2 (Xnr2). In this report, we have followed cells expressing Xbra in the presumptive trunk and tail at the gastrula stage, and find that they fate to presumptive somite, but not to ventrolateral mesoderm of the tailbud embryo. From this, we speculate that the boundary between the Xbra- and Xnr2-expressing cells at gastrula corresponds to a future tissue boundary. In further experiments, we show that the level of mitogen-activated protein kinase (MAPK) activation is polarized along the animal/vegetal axis, with the Xnr2-expressing cells in the vegetal marginal zone having no detectable activated MAPK. We show that inhibition of MAPK activation in Xenopus animal caps results in the conversion of Xnr2 from a dorsal mesoderm inducer to a ventral mesoderm inducer, supporting a role for Xnr2 in induction of ventral mesoderm.