MUSCLE SATELLITE CELL--REACTION TO DISEASE AND TRAUMA

成肌细胞转移被认为是一种基因互补的方法 用于原发性肌病和补充蛋白质， 体循环该系统还提供了一个独特的机会， 评估肌肉发育和再生。建议研究 使用胎儿宿主和稳定的成肌细胞转移系统的细胞生物学 标记成肌细胞。利用胎儿宿主将克服这些问题 这是成肌细胞转移到更成熟宿主中所固有的（即，问题 供体成肌细胞的递送、分散和整合成显著的 肌纤维的数量和免疫排斥的问题），如在 注射免疫缺陷胎儿的小肌肉 宿主将经历广泛的从头肌管形成， 组织不良的结缔组织。这个百万应该导致 产生大量广泛分布的镶嵌肌管 (with不同频率的可识别的供体成肌细胞）， 通过肌肉创伤诱导成肌细胞 并入含有其全部肌纤维补体的肌肉中。 这将允许评估注射的成肌细胞的命运， 以确定它们的后代是否保持为肌卫星 细胞，能够参与肌纤维生长和再生， 来源于供体的肌细胞核长期存活的能力， 继续生产一种独特的结构蛋白质产品。问题 成肌细胞向特定细胞谱系的定型将通过 从胚胎肌肉中转移稳定标记的成肌细胞， 肌卫星细胞从成人肌肉转化为发育中的胎儿肌肉。我们将 还强调了使用成肌细胞转移作为一种方法的潜在益处 基因互补的替代肌营养不良蛋白进入 胎儿mdx小鼠的肌营养不良蛋白缺陷型肌纤维。捐赠频率 在mdx小鼠中抗肌萎缩蛋白阳性镶嵌肌纤维中宿主肌核 以及肌营养不良蛋白替代物对修饰 评估肌营养不良蛋白缺陷肌肉的变性-再生。 将采用免疫组织化学和光学显微镜进行评价 和扫描共聚焦显微镜，并与蛋白质印迹抗体， 抗肌营养不良蛋白和抗肌球蛋白或诱导的抗体。结果 这些实验将提供重要的新信息， 成肌细胞转移系统将为未来使用 该系统用于治疗目的并作为评估许多 肌肉发育的细胞生物学方面。

Myoblast transfer has been suggested as a method of gene complementation for primary myopathics and to supplement proteins that may be absent in the systemic circulation. This system also offers a unique opportunity to evaluate muscle development and regeneration. It is proposed to study the cell biology of the myoblast transfer system using fetal hosts and stably marked myoblasts. The use of fetal hosts will overcome the problems inherent in myoblast transfer into more mature hosts (i.e., problems of delivery, dispersal and integration of donor myoblasts into a significant number of myofibers and problems of immune rejection), as at the time of injection the small developing muscles of the immune-incompetent fetal hosts will be undergoing extensive de novo myotube formation and will have poorly organized connective tissue. This millieu should result in the production of a significant number of widely dispersed mosaic myotubes (with different frequencies of identifiable donor myoblast), without resorting to the muscle traumatization required to induce myoblast incorporation into muscles containing their full complement of myofibers. This will permit evaluation of the fate of the injected myoblast, determining whether some of their progeny are maintained as myosatellite cell, capable of participating in myofiber growth and regeneration and of the ability of donor-derived myonuclei to survive long-term and to continue to produce a unique structural protein product. The question of myoblast commitment to specific cell lineages will then be addressed by transferring stably marked myoblasts from embryonic muscle and myosatellite cells from adult muscle into developing fetal muscle. We will also address -the potential benefit of using myoblast transfer as a method of gene complementation for the replacement of dystrophin into the dystrophin-deficient myofibers of fetal mdx mice. The frequency of donor to host myonuclei in dystrophin-positive mosaic myofibers in the mdx mice and the effectiveness of dystrophin replacement on modifying the degeneration-regeneration of dystrophin deficient muscle will be assessed. Evaluations will be made with immunohistochemistry, and light microscopy and scanning confocal microscopy, and with Western blotting with antibody against dystrophin and antibodies against myosin or induction. The results of the proposed experiments will provide significant new information about the myoblast transfer system that will be open the way for future use of this system for therapeutic purposes and as a tool for evaluating many aspects of the cell biology of muscle development.