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通过SDS聚丙烯酰胺凝胶电泳和电子显微镜对发育中的鸡心脏收缩蛋白进行分析。

An analysis of contractile proteins in developing chick heart by SDS polyacrylamide gel electrophoresis and electron microscopy.

作者信息

Lim S S, Woodroofe M N, Lemanski L F

出版信息

J Embryol Exp Morphol. 1983 Oct;77:1-14.

PMID:6655428
Abstract

Chick heart development was studied using transmission electron microscopy and SDS-polyacrylamide gel electrophoresis in combination with densitometry. Myosin heavy chain, alpha-actinin, actin and tropomyosin accumulations were analysed in developing hearts from preheartbeat stage 9 (Hamburger-Hamilton staging series) through 2 days after hatching. At the preheartbeat stage, electron microscopy revealed a significant number of thin filaments scattered throughout the cytoplasm of the myoblasts; however, very few thick filaments were seen. There was no obvious association between the two filament types. SDS-polyacrylamide tube gels of heart muscle homogenates demonstrated the presence of all five proteins in hearts at the preheartbeat stage. Further analyses of the proteins by gel densitometry indicated that both actin and myosin accumulated rapidly during heart development while alpha-actinin and tropomyosin levels remained relatively static. Our results show that detectable quantities of myosin heavy chain, alpha-actinin, actin and tropomyosin accumulate in myocardial cells prior to the appearance of myofibrils and initiation of the contractile function.

摘要

利用透射电子显微镜以及结合光密度测定法的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对鸡心脏发育进行了研究。分析了从心跳前期9阶段(汉堡-汉密尔顿分期系列)到孵化后2天发育中心脏中肌球蛋白重链、α-辅肌动蛋白、肌动蛋白和原肌球蛋白的积累情况。在心跳前期,电子显微镜显示大量细肌丝散布在成肌细胞的整个细胞质中;然而,可见的粗肌丝很少。两种肌丝类型之间没有明显的关联。心肌匀浆的十二烷基硫酸钠-聚丙烯酰胺管状凝胶显示在心跳前期的心脏中存在所有五种蛋白质。通过凝胶光密度测定法对这些蛋白质进行的进一步分析表明,在心脏发育过程中肌动蛋白和肌球蛋白均迅速积累,而α-辅肌动蛋白和原肌球蛋白水平保持相对稳定。我们的结果表明,在肌原纤维出现和收缩功能启动之前,心肌细胞中就积累了可检测量的肌球蛋白重链、α-辅肌动蛋白、肌动蛋白和原肌球蛋白。

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