Expression profiling and identification of novel genes involved in myogenic differentiation
Expression profiling and identification of novel genes involved in myogenic differentiation
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DOI：
10.1096/fj.03-0568fje
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发表时间：
2004-02-01
期刊：
影响因子：
4.8
通讯作者：
Beggs, Alan H.
中科院分区：
文献类型：
--
作者：
Tomczak, Kinga K.;Marinescu, Voichita D.;Beggs, Alan H.
骨骼肌分化是一个复杂的，高度协调的过程，依赖于精确的时间基因表达模式。为了更好地理解这种级联的转录事件，我们使用表达谱分析在分化C2 C12成肌细胞的12天时间过程中的基因表达。聚类分析特定的时间顺序的微阵列实验分类2895个基因和ESTs与不同的表达水平之间的增殖和分化的细胞成22个簇与不同的表达模式在肌发生。通过实时定量RT-PCR和/或Western印迹和免疫荧光法独立地确认了几个已知和新基因的表达模式。MyoD和MEF家族成员表现出独特的表达动力学，与细胞周期退出调节因子高度协调。在细胞周期退出期间具有峰值表达水平的基因中有Vcam 1、Itgb 3、Itga 5、Vcl以及Ptger 4，Ptger 4是先前与肌生成过程无关的基因。一个有趣的未表征的转录，是高度诱导肌生成过程中编码几个免疫球蛋白重复序列的肌联蛋白，一个大的肌节蛋白的序列相似性。这些数据集确定了许多额外的未表征的转录本，这些转录本可能在肌细胞增殖和分化中发挥重要作用，并为与表达各种肌病疾病相关突变基因的C2 C12细胞进行比较提供了基线。
Skeletal muscle differentiation is a complex, highly coordinated process that relies on precise temporal gene expression patterns. To better understand this cascade of transcriptional events, we used expression profiling to analyze gene expression in a 12-day time course of differentiating C2C12 myoblasts. Cluster analysis specific for time-ordered microarray experiments classified 2895 genes and ESTs with variable expression levels between proliferating and differentiating cells into 22 clusters with distinct expression patterns during myogenesis. Expression patterns for several known and novel genes were independently confirmed by real-time quantitative RT-PCR and/or Western blotting and immunofluorescence. MyoD and MEF family members exhibited unique expression kinetics that were highly coordinated with cell-cycle withdrawal regulators. Among genes with peak expression levels during cell cycle withdrawal were Vcam1, Itgb3, Itga5, Vcl, as well as Ptger4, a gene not previously associated with the process of myogenesis. One interesting uncharacterized transcript that is highly induced during myogenesis encodes several immunoglobulin repeats with sequence similarity to titin, a large sarcomeric protein. These data sets identify many additional uncharacterized transcripts that may play important functions in muscle cell proliferation and differentiation and provide a baseline for comparison with C2C12 cells expressing various mutant genes involved in myopathic disorders.