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Functional Gene Expression Analysis of Tissue-Specific Isoforms of Mef2c

Cellular and Molecular Neurobiology volume 32, pages 129–139 (2012)Cite this article

Abstract

Mef2c contains three alternative exons and generates six Mef2c isoforms in mice. Mef2c α1β isoforms are expressed in neuronal tissues, α2 isoforms are expressed in muscle, and α1 isoforms are expressed in many other tissues. The γ region inclusion and skipped isoforms are present in equal amounts in many tissues. In this study, differences in the transcriptional activities of each tissue-specific isoform of Mef2c in neuronal cells were examined. Using an MEF2-responsive reporter, exon β-dependent transactivation was found in neuronal cells, as well as in other cell lines previously described. Microarray analysis was used to examine the transcriptional activities of each Mef2c isoform and to assess differences in endogenous gene expression induced by the different isoforms. The results showed significant gene expression changes due to overexpression of Mef2c isoforms in both an isoforms-dependent and -independent manner.

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Acknowledgments

We thank Assoc. Prof. J. Sese (Tokyo Institute of Technology) for advice on the microarray analysis. We thank Prof. E. N. Olson (UT Southwestern) for the pGL3-TATA-desmef2 plasmid.

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Affiliations

  1. School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa, 923-1292, Japan

    Yoshiharu Sekiyama, Hitoshi Suzuki & Toshifumi Tsukahara

  2. Center for Nano Materials and Technology, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa, 923-1292, Japan

    Hitoshi Suzuki & Toshifumi Tsukahara

Authors
  1. Yoshiharu Sekiyama
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  2. Hitoshi Suzuki
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  3. Toshifumi Tsukahara
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Correspondence to Toshifumi Tsukahara.

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Supplementary material 1 (XLS 88 kb)

10571_2011_9743_MOESM2_ESM.tiff

Supplemental Fig. 1. Dose-dependent luciferase induction of Mef2c isoforms. Neuro2A cells were transfected with 200 ng of pGL-TATA-desmef2 and 20 ng of pRL-Tk together with 2.56 ng, 6.4 ng, 16 ng, 40 ng, 100 ng or 250 ng of the Mef2c expression plasmids. Firefly luciferase activity was normalized to Renilla luciferase activity. (TIFF 90 kb)

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Sekiyama, Y., Suzuki, H. & Tsukahara, T. Functional Gene Expression Analysis of Tissue-Specific Isoforms of Mef2c. Cell Mol Neurobiol 32, 129–139 (2012). https://doi.org/10.1007/s10571-011-9743-9

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  • DOI: https://doi.org/10.1007/s10571-011-9743-9

Keywords

  • Alternative splicing
  • Mef2c isoforms
  • Transcriptional activity
  • Microarray analysis