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Expression profiling and identification of novel genes involved in myogenic differentiation
Author(s) -
Tomczak Kinga K.,
Marinescu Voichita D.,
Ramoni Marco F.,
Sanoudou Despina,
Montanaro Federica,
Han Mei,
Kunkel Louis M.,
Kohane Isaac S.,
Beggs Alan H.
Publication year - 2004
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.03-0568fje
Subject(s) - myogenesis , biology , myod , c2c12 , gene expression profiling , gene , gene expression , genetics , myogenin , microarray analysis techniques , cellular differentiation , microbiology and biotechnology
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 , Vc1 , 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.