Cripe L, Morris E, Fulton A B
Department of Biochemistry, University of Iowa, Iowa City 52242.
Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2724-8. doi: 10.1073/pnas.90.7.2724.
The mRNAs for some cytoskeletal proteins are localized, suggesting that mRNA for these proteins may concentrate at sites appropriate for assembly. To test this hypothesis, we observed vimentin mRNA in developing chicken muscle cultures by in situ hybridization with a digoxigenin-labeled DNA probe to vimentin, detected by confocal microscopy using fluorescent anti-digoxigenin antibody. This method has submicrometer resolution. In developing muscle, vimentin mRNA was bipolar in young myoblasts, somewhat perinuclear in elongated myoblasts and spread fibroblasts, and diffuse in young and developing myotubes. In mature myotubes, vimentin mRNA occurred at costameres with vimentin protein. Localization of mRNA may prove important for assembling and maintaining differentiated cytoskeletal structures, as it is for organizing the embryo.
一些细胞骨架蛋白的信使核糖核酸（mRNA）是定位的，这表明这些蛋白质的mRNA可能集中在适合组装的位点。为了验证这一假设，我们通过用抗地高辛配体抗体标记的地高辛配体标记的波形蛋白DNA探针原位杂交，在发育中的鸡肌肉培养物中观察波形蛋白mRNA，并用荧光抗地高辛配体抗体通过共聚焦显微镜检测。该方法具有亚微米分辨率。在发育中的肌肉中，波形蛋白mRNA在年轻的成肌细胞中呈双极分布，在伸长的成肌细胞和伸展的成纤维细胞中有点呈核周分布，在年轻和发育中的肌管中呈弥散分布。在成熟的肌管中，波形蛋白mRNA与波形蛋白一起出现在肌节旁。mRNA的定位对于组装和维持分化的细胞骨架结构可能很重要，就像它对于胚胎发育的组织作用一样。
