首页> 外文OA文献 >Molecular cloning of up-regulated cytoskeletal genes from regenerating skeletal muscle: potential role of myocyte enhancer factor 2 proteins in the activation of muscle-regeneration-associated genes.
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Molecular cloning of up-regulated cytoskeletal genes from regenerating skeletal muscle: potential role of myocyte enhancer factor 2 proteins in the activation of muscle-regeneration-associated genes.

机译:从再生骨骼肌上调细胞骨架基因的分子克隆:肌细胞增强因子2蛋白在与肌肉再生相关的基因的激活中的潜在作用。

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摘要

A subtractive hybridization and cloning strategy was used to identify genes that are up-regulated in regenerating compared with normal skeletal muscle. The gastrocnemius muscle of CD1 mice was injected with a myotoxic agent (BaCl2). A cDNA library was constructed from the regenerating muscle, and was screened with subtracted probes enriched in genes up-regulated during regeneration. Cofilin and vimentin cDNA clones were isolated. Both cofilin and vimentin were demonstrated to be overexpressed in regenerating compared with non-regenerating muscle (17-fold and 19-fold induction respectively). Cofilin and vimentin mRNAs also exhibited an increased expression in C2C12 myoblasts and a decreased expression in differentiated myotubes. Analysis of the regeneration-induced vimentin enhancer/promoter region revealed a consensus binding site for the myocyte enhancer factor 2 (MEF2) transcription factors. Electrophoretic mobility-shift assays and in vivo reporter assays revealed that MEF2 DNA-binding activity and transcriptional activation are increased in regenerating skeletal muscle, indicating that they may play a role in the activation of muscle genes during regeneration. These data suggest that both cofilin (an actin-regulatory protein) and vimentin (an intermediate filament) may be key components of the cytoskeletal reorganization that mediates muscle cell development and adult skeletal-muscle repair.
机译:减性杂交和克隆策略用于鉴定与正常骨骼肌相比在再生中上调的基因。 CD1小鼠的腓肠肌注射了肌毒性剂(BaCl2)。从再生的肌肉中构建了一个cDNA文库,并用富含在再生过程中上调的基因的扣除探针进行筛选。分离出Cofilin和波形蛋白cDNA克隆。与非再生肌肉相比,cofilin和波形蛋白都被证明在再生中过表达(诱导分别为17倍和19倍)。 Cofilin和波形蛋白mRNA在C2C12成肌细胞中也表达增加,而在分化的肌管中表达减少。对再生诱导的波形蛋白增强子/启动子区域的分析显示了心肌细胞增强子因子2(MEF2)转录因子的共有结合位点。电泳迁移率迁移测定和体内报告基因测定表明,MEF2 DNA结合活性和转录激活在再生骨骼肌中增加,表明它们可能在再生过程中对肌肉基因的激活起作用。这些数据表明cofilin(肌动蛋白调节蛋白)和波形蛋白(中间丝)可能是介导肌肉细胞发育和成年骨骼肌修复的细胞骨架重组的关键成分。

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