Analysis of altered gene expression in rat soleus muscle atrophied by disuse
Analysis of altered gene expression in rat soleus muscle atrophied by disuse
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DOI：
10.1002/jcb.1248
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发表时间：
2001-01-01
影响因子：
4
通讯作者：
Dechesne, CA
中科院分区：
文献类型：
--
作者：
Cros, N;Tkatchenko, AV;Dechesne, CA
本研究涉及对后肢悬吊（HS）后萎缩的大鼠比目鱼肌中表达发生改变的基因进行全面分析。 HS 肌肉卸载是肌肉废用的常见模型，尤其影响反重力慢肌，例如比目鱼肌。基于抑制消减杂交技术的 cDNA 克隆策略导致构建了两个标准化比目鱼肌 cDNA 文库，这两个文库以相反方向进行消减，即用对照 cDNA 减去萎缩的比目鱼肌 cDNA，反之亦然。对两个文库的差异筛选发现，除了 HS 后仅在比目鱼肌中表达的 2X 和 2B 肌球蛋白重链基因外，还有 34 个基因在 HS 比目鱼肌中表达发生改变，其中包括 11 个新的 cDNA。通过反向 Northern 印迹和经典 Northern 印迹分析对基因上调和下调进行定量。 25 个具有已知功能的基因分为七个重要的功能类别。每个类别内基因改变的同质性为揭示肌肉萎缩表型中隐含的细胞事件的相互作用提供了一些线索。特别是，我们的结果表明，慢肌和快肌成分平衡、蛋白质合成/分泌途径和细胞外基质/细胞骨架轴的调节可能是肌肉萎缩的关键分子机制。此外，新型 cDNA 的克隆强调了所选技术方法的效率，并为进一步破译肌肉萎缩的分子机制提供了新的可能性。 J.细胞。生物化学。 83: 508-519, 2001。(C) 2001 Wiley-Liss, Inc.
The present study involved a global analysis of genes whose expression was modified in rat soleus muscle atrophied after hindlimb suspension (HS). HS muscle unloading is a common model for muscle disuse that especially affects antigravity slow-twitch muscles such as the soleus muscle. A cDNA cloning Strategy, based on suppression subtractive hybridization technology, led to the construction of two normalized soleus muscle cDNA libraries that were subtracted in opposite directions, i.e., atrophied soleus muscle cDNAs subtracted by control cDNAs and vice versa. Differential screening of the two libraries revealed 34 genes with altered expression in HS soleus muscle, including 11 novel cDNAs, in addition to the 2X and 2B myosin heavy chain genes expressed only in soleus muscles after HS. Gene up- and down-regulations were quantified by reverse Northern blot and classical Northern blot analysis. The 25 genes with known functions fell into seven important functional categories. The homogeneity of gene alterations within each category gave several clues for unraveling the interplay of cellular events implied in the muscle atrophy phenotype. In particular, our results indicate that modulations in slow- and fast-twitch-muscle component balance, the protein synthesis/secretion pathway, and the extracellular matrix/cytoskeleton axis are likely to be key molecular mechanisms of muscle atrophy. In addition, the cloning of novel cDNAs underlined the efficiency of the chosen technical approach and gave novel possibilities to further decipher the molecular mechanisms of muscle atrophy. J. Cell. Biochem. 83: 508-519, 2001. (C) 2001 Wiley-Liss, Inc.