Distribution of myosin heavy chain mRNA in embryonic muscle tissue visualized by ultrastructural in situ hybridization.
Distribution of myosin heavy chain mRNA in embryonic muscle tissue visualized by ultrastructural in situ hybridization.
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通过超微结构原位杂交可视化胚胎肌肉组织中肌球蛋白重链 mRNA 的分布。
DOI：
10.1016/0012-1606(91)90054-7
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发表时间：
1991
影响因子：
2.7
通讯作者：
Billings-Gagliardi,S
中科院分区：
文献类型：
--
作者：
Pomeroy,ME;Lawrence,JB;Singer,RH;Billings-Gagliardi,S
我们利用高分辨率原位杂交技术，在完整的鸡胚肌细胞中定位了肌球蛋白重链(MHC)mRNAs。在洗涤剂处理之前，肌肉块被醛固定，并与生物标记的cdna探针杂交，随后是胶体金标记的抗体，然后嵌入。通过对实验样品和四种不同类型的对照样品的电子显微照片进行广泛的定量比较，确定标记代表MHC mRNA。MHC mRNA主要定位于14日龄雏鸡胸肌细胞的周边区域，在那里发现了大多数发育中的肌原纤维。MHC mRNAs与直径在4~10 nm之间的非肌原纤维细胞骨架丝联系在一起。它们通常平行于细胞的纵轴定向。超微结构方法的分辨率使我们能够证明可视化的mRNA分子与肌丝没有直接关联，这表明从这些信息中读取的新生链不会在翻译的同时直接组装成肌丝。
We have localized myosin heavy chain (MHC) mRNAs in cells of intact embryonic chick muscle using high resolutionin situhybridization. Blocks of muscle were aldehyde-fixed prior to detergent treatment and hybridized with a biotinated cDNA probe, followed by colloidal gold-labeled antibodies, before embedment. Labeling was determined to represent MHC mRNA by extensive quantitative comparisons of electron micrographs from experimental and four different types of control samples. MHC mRNA was localized primarily to peripheral regions of 14-day chick pectoral muscle cells, where the majority of developing myofibrils were found. MHC mRNAs were consistently associated with the nonmyofibrillar cytoskeletal filaments which had diameters ranging from 4 to 10 nm. They were often oriented parallel to the longitudinal axis of the cell. The resolution of the ultrastructural approach allowed us to demonstrate that the mRNA molecules visualized were not directly associated with myofilaments, suggesting that nascent chains read from those messages do not assemble directly into myofilaments simultaneous with translation.