NORMAL AND PATHOLOGIC MUSCLE IN TISSUE CULTURE

我们打算研究发育调节的同工酶转换， 磷酸化、磷酸果糖激酶、腺苷酸脱氨酶和 人非神经支配和神经支配肌肉中磷酸甘油酸酯酶 从正常和遗传缺陷的肌肉中培养， 免疫细胞化学、组织化学和电泳技术。 这些 研究将有助于更好地了解调节同工酶的因素 过渡和代谢后果，当这种过渡失败时， 是由于基因缺陷造成的。 我们还建议发展永久性的人力资源， 来自正常和遗传缺陷肌肉的成肌细胞系， 转化来自初级肌的成肌细胞， 温度敏感突变型SV40病毒。 成肌细胞转化体，不像 来自原代组织的培养细胞不会衰老， 成纤维细胞污染，并可诱导分化 温度变化 这些细胞系将提供无限的 用于研究正常和遗传缺陷人类肌肉的大样本. 早期的分化状态，并将允许更详细的分析 在分子水平上对人类肌肉发育的影响。

We intend to study developmentally regulated isoenzyme transitions of phosphorylate, phosphofructokinase, adenylate deaminase and phosphoglycerate mutase in uninnervated and innervated human muscle cultures from normal and genetically deficient muscle using immunocytochemical, histochemical and electrophoretic techniques. These studies will lead to a better understanding of factors regulating isoenzyme transitions and the metabolic consequences, when such transitions fail to occur due to genetic defects. We also propose to develop permanent human myoblast cell lines from normal and genetically-deficient muscle by transforming myoblasts derived from primary muscle with a temperature-sensitive mutant SV40 virus. Myoblast transformants, unlike cultured cells from primary tissues do not become senescent, are free of fibroblast contamination, and can be induced to differentiate by temperature shift. These cell lines will provide unlimited supplies of large samples for studying normal and genetically deficient human muscle at early states of differentiation, and will permit a more detailed analysis of human muscle development at the molecular level.