Impact of denervation-induced muscle atrophy on housekeeping gene expression in mice

Authors

  • Reiko Nakao PhD,

    1. Biological Clock Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan
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  • Saori Yamamoto MS,

    1. Biological Clock Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan
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  • Yuki Yasumoto BSc,

    1. Biological Clock Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan
    2. Department of Applied Biological Science, Graduate School of Science and Technology, Tokyo University of Science, Chiba, Japan
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  • Koji Kadota PhD,

    1. Agricultural Bioinformatics Research Unit, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan
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  • Katsutaka Oishi PhD

    Corresponding author
    1. Biological Clock Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan
    2. Department of Applied Biological Science, Graduate School of Science and Technology, Tokyo University of Science, Chiba, Japan
    3. Department of Medical Genome Sciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba, Japan
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  • This study was supported by operational subsidies from AIST and KAKENHI [Grant-in-Aid for Scientific Research (C) 24620015 to R.N.] from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan.

ABSTRACT

Introduction: Immobilization induced by experimental denervation leads to rapid and progressive alterations in structural and biochemical properties of skeletal muscle. Real-time reverse transcription-polymerase chain reaction (RT-PCR) is a popular method of elucidating the molecular mechanisms involved in muscle atrophy. Identification of suitable reference genes that are not affected by experimental conditions is a critical step in accurate normalization of real-time RT-PCR. Methods: We investigated the impact of denervation-induced muscle atrophy for 2 weeks on the expression of common housekeeping genes. Results: Denervation differentially affected the expression levels of these genes. RefFinder software identified TATA box binding protein (Tbp) as the most stable gene and showed that the stability of glyceraldehyde-3-phosphate dehydrogenase (Gapdh) and hypoxanthine guanine phosphoribosyl transferase (Hprt) genes was low, even though they are widely used for normalization. Conclusions: The appropriate reference gene for normalization of genes of interest in denervated muscle is Tbp. Muscle Nerve 51: 276–281, 2015

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