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Cellular localization of muscle and nonmuscle actin mRNAs in chicken primary myogenic cultures: the induction of alpha-skeletal actin mRNA is regulated independently of alpha-cardiac actin gene expression

机译：鸡原代成肌培养物中肌肉和非肌肉肌动蛋白mRNA的细胞定位：α骨骼肌肌动蛋白mRNA的诱导独立于α心脏肌动蛋白基因表达的调控

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摘要

Specific DNA fragments complementary to the 3' untranslated regions of the beta-, alpha-cardiac, and alpha-skeletal actin mRNAs were used as in situ hybridization probes to examine differential expression and distribution of these mRNAs in primary myogenic cultures. We demonstrated that prefusion bipolar-shaped cells derived from day 3 dissociated embryonic somites were equivalent to myoblasts derived from embryonic day 11-12 pectoral tissue with respect to the expression of the alpha-cardiac actin gene. Fibroblasts present in primary muscle cultures were not labeled by the alpha-cardiac actin gene probe. Since virtually all of the bipolar cells express alpha-cardiac actin mRNA before fusion, we suggest that the bipolar phenotype may distinguish a committed myogenic cell type. In contrast, alpha-skeletal actin mRNA accumulates only in multinucleated myotubes and appears to be regulated independently from the alpha-cardiac actin gene. Accumulation of alpha- skeletal but not alpha-cardiac actin mRNA can be blocked by growth in Ca2+-deficient medium which arrests myoblast fusion. Thus, the sequential appearance of alpha-cardiac and then alpha-skeletal actin mRNA may result from factors that arise during terminal differentiation. Finally, the beta-actin mRNA was located in both fibroblasts and myoblasts but diminished in content during myoblast fusion and was absent from differentiated myotubes. It appears that in primary myogenic cultures, an asynchronous stage-dependent induction of two different alpha-striated actin mRNA species occurs concomitant with the deinduction of the nonmuscle beta-actin gene.
机译：与β，α-心脏和α-骨架肌动蛋白mRNA的3'非翻译区互补的特定DNA片段用作原位杂交探针，以检查这些mRNA在原代成肌培养物中的差异表达和分布。我们证明，就α-心脏肌动蛋白基因的表达而言，源自第3天解离的胚胎节的融合前双极型细胞与源自胚胎第11-12天的胸膜组织的成肌细胞相同。存在于原代肌肉培养物中的成纤维细胞没有被α-心脏肌动蛋白基因探针标记。由于实际上所有双极细胞都在融合前表达α-心脏肌动蛋白mRNA，因此我们建议双极表型可以区分特定的成肌细胞类型。相反，α-骨骼肌肌动蛋白mRNA仅在多核肌管中蓄积，并且似乎独立于α-心脏肌动蛋白基因受到调节。在缺乏Ca2 +的培养基中可阻止成肌细胞融合，阻止α-骨骼肌而不是α-心脏肌动蛋白mRNA的积累。因此，α心脏继而出现的α骨骼肌肌动蛋白mRNA的顺序出现可能是由终末分化过程中出现的因素引起的。最后，β-肌动蛋白mRNA位于成纤维细胞和成肌细胞中，但在成肌细胞融合过程中含量减少，并且分化的肌管中不存在。似乎在原发性肌源性培养中，伴随着非肌肉β-肌动蛋白基因的缺失，发生了两个不同的α-横纹肌动蛋白mRNA种类的异步阶段依赖性诱导。

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