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Cyclosporin A treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration

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Abstract

The molecular signaling pathway linked to muscle regeneration has not yet been identified. Previously, we demonstrated that mice treated with cyclosporin A (CsA), a calcineurin inhibitor, failed to regenerate normally after muscle damage. Using reverse transcription (RT)-PCR, Western blot and immunohistochemical analysis, we investigated whether the amounts of nuclear factor of activated T cells (NFAT), myocyte-enhancer factor 2 (MEF2), the MyoD family, Id-1, and Smad3 change in the regenerating muscle after CsA treatment. Adult male ICR mice were subjected to a bupivacaine injection into the tibialis anterior muscle, and were treated with either CsA (25 mg/kg) or vehicle once daily. They were killed at 1, 2, 4, 6, 9 and 14 days post injury. RT-PCR analysis did not show a significant difference in MEF2s, MyoD and myogenin mRNA levels in the regenerating muscle in either placebo- and CsA-administered mice. In contrast, a significant increase in MRF4 mRNA was seen in CsA-administered mice compared to the placebo-treated mice at 4 and 9 days post surgery. In CsA-treated mice, the level of Id1 mRNA was elevated at day 9 relative to the placebo-treated mice. After 6 days, the CsA-treated mice possessed more abundant proliferating cell nuclear antigen (PCNA) and cyclin D1 protein in many satellite cells and/or myoblast-like cells in the regenerating muscle. The amount of myostatin, TGF-β2 and Smad3 mRNA and proteins was increased more markedly in the mice treated with CsA. After 9 days, many satellite cells and/or myoblasts showed apparent co-localization of both MyoD and Smad3 in CsA-, but not in placebo-, treated mice. Our results demonstrated that CsA treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration in vivo.

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Acknowledgements

This work was supported by a research Grant-in-Aid for Young Scientists B (nos. 13780029 and 15700423) from the Ministry of Education, Science, Sports and Culture of Japan.

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Authors and Affiliations

  1. Department of Legal Medicine, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kawaramachi-hirokoji, Kamigyo-ku, 602-8566, Kyoto, Japan

    Kunihiro Sakuma, Ryuta Nakao, Shuichiro Inashima, Miyuki Hirata & Masahiro Yasuhara

  2. Department of First Medicine, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kawaramachi-hirokoji, Kamigyo-ku, 602-8566, Kyoto, Japan

    Wataru Aoi

  3. Department of Biochemistry, Mie University Faculty of Medicine, Tsu, 514-8507, Mie, Japan

    Takahiko Fujikawa

  4. Department of Biology, Kyoto Prefectural University of Medicine, Nishitakatsukasa, Taisyougun, Kitaku, 603-8334, Kyoto, Japan

    Mamoru Sano

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  1. Kunihiro Sakuma
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  2. Ryuta Nakao
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  3. Wataru Aoi
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  8. Masahiro Yasuhara
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Correspondence to Kunihiro Sakuma.

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Sakuma, K., Nakao, R., Aoi, W. et al. Cyclosporin A treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration. Acta Neuropathol 110, 269–280 (2005). https://doi.org/10.1007/s00401-005-1049-x

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  • DOI: https://doi.org/10.1007/s00401-005-1049-x

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