The role of Ena/VASP proteins in muscle development

描述(由申请人提供)：Ena/Vasp蛋白在肌动蛋白细胞骨架的组织中的作用以及对细胞运动和轴突生长的调节已经得到很好的证实。然而，它们在胚胎肌肉和肌肉前体中的表达表明，在发育过程中肌肉的组织和分化具有额外的作用。我建议使用非洲爪哇胚胎和培养的原代鸡骨骼肌细胞来研究Ena/Vasp蛋白在肌肉发育过程中的时空表达和亚细胞定位。这些蛋白的缺失突变体的构建和表达将有助于确定针对特定亚细胞定位所需的结构域，并为深入了解Ena/Vasp在肌小体中功能的分子基础提供依据。利用显性负性结构和吗啉寡核苷酸特异性抑制Ena/Vasp蛋白的功能和表达，我将研究这种缺失在发育过程中对细胞骨架和细胞结构和功能的影响。这些研究将阐明Ena/Vasp蛋白在骨骼肌发育中的作用，并可能产生关于肌肉适应和疾病的分子和细胞基础的新信息。

DESCRIPTION (provided by applicant): The role of Ena/VASP proteins in the organization of the actin cytoskeleton and the regulation of cell motility and axon outgrowth has been well established. However, their expression in embryonic muscle and muscle precursors suggests an additional role in the organization and differentiation of muscles during development. I propose to examine the spatial and temporal expression, and the subcellular localization of Ena/VASP proteins during muscle development, using both Xenopus embryos and cultured primary chick skeletal muscle cells. Construction and expression of deletion mutants of these proteins will allow determination of the domains necessary for targeting to specific subcellular Iocalizations, and provide insights into the molecular basis of Ena/VASP function in the myotome. Using a dominant negative construct and morpholino oligonucleotides to specifically inhibit Ena/VASP protein function and expression, I will investigate the effects of this loss on cytoskeletal and cellular structure and function during development. These investigations will elucidate the role of Ena/VASP proteins in skeletal muscle development, and may produce novel information concerning the molecular and cellular basis of muscle adaptation and disease.