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Regulation of the Expression of the Myosin Heavy Chain (MYH) Gene myh14 in Zebrafish Development

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Abstract

The human sarcomeric myosin heavy chain gene MYH14 contains an intronic microRNA, miR-499. Our previous studies demonstrated divergent genomic organization and expression patterns of myh14/miR-499 among teleosts; however, the regulatory mechanism is partly known. In this study, we report the regulation of myh14 expression in zebrafish, Danio rerio. Zebrafish myh14 has three paralogs, myh14-1, myh14-2, and myh14-3. Detailed promoter analysis suggested that a 5710-bp 5′-flanking region of myh14-1 and a 5641-bp region of myh14-3 contain a necessary regulatory region to recapitulate specific expression during embryonic development. The 5′-flanking region of zebrafish myh14-1 and its torafugu ortholog shared two distal and a single proximal conserved region. The two distal conserved regions had no effect on zebrafish myh14-1 expression, in contrast to torafugu expression, suggesting an alternative regulatory mechanism among the myh14 orthologs. Comparison among the 5′-flanking regions of the myh14 paralogs revealed two conserved regions. Deletion of these conserved regions significantly reduced the promoter activity of myh14-3 but had no effect on myh14-1, indicating different cis-regulatory mechanisms of myh14 paralogs. Loss of function of miR-499 resulted in a marked reduction in slow muscle fibers in embryonic development. Our study identified different cis-regulatory mechanisms controlling the expression of myh14/miR-499 and an indispensable role of miR-499 in muscle fiber–type specification in zebrafish.

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Funding

This study was partly supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science and by the start-up package from Texas A&M International University.

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  1. Sharmin Hasan

    Present address: Department of Biology and Chemistry, Texas A&M International University, 5201 University Blvd., Laredo, TX, 78041, USA

Authors and Affiliations

  1. Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo, 113-8657, Japan

    Sharmin Hasan, Shuichi Asakawa & Shigeharu Kinoshita

  2. School of Marine Bioscience, Kitasato University, Minami-ku, Sagamihara, Kanagawa, 252-0373, Japan

    Shugo Watabe

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  1. Sharmin Hasan
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  2. Shuichi Asakawa
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  4. Shigeharu Kinoshita
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Contributions

S.H. and S.K. conceived and designed the study and performed the experiments. S.H. performed the data acquisition and interpretation. S.A. and S.W. participated in research design and coordination. S.H. wrote the manuscript.

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Correspondence to Sharmin Hasan.

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Hasan, S., Asakawa, S., Watabe, S. et al. Regulation of the Expression of the Myosin Heavy Chain (MYH) Gene myh14 in Zebrafish Development. Mar Biotechnol 23, 821–835 (2021). https://doi.org/10.1007/s10126-021-10066-z

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