A new monoclonal antibody, 4F2, specific for the oligodendroglial cell lineage, recognizes ATP-dependent RNA helicase Ddx54: Possible association with myelin basic protein

Authors

  • Toshiyuki Ueki,

    1. Department of Neuro-Glia Cell Biology, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan
    2. Tsumura Laboratories, Tsumura & Co., Ibaraki, Japan
    3. Department of Biochemistry, Graduate School of Pharmaceutical Sciences, Meiji Pharmaceutical University, Tokyo, Japan
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    • T. Ueki and Y. Tsuruo contributed equally to this work.

  • Yoshihiro Tsuruo,

    1. Department of Anatomy and Cell Biology, Wakayama Medical University School of Medicine, Wakayama, Japan
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    • T. Ueki and Y. Tsuruo contributed equally to this work.

  • Yuta Yamamoto,

    1. Department of Anatomy and Cell Biology, Wakayama Medical University School of Medicine, Wakayama, Japan
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  • Kazunori Yoshimura,

    1. Department of Neuro-Glia Cell Biology, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan
    2. Nihon Institute of Medical Sciences, Saitama, Japan
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  • Hiromi Takanaga,

    1. Cell Lineage Modulation, Center for Developmental Biology, Kobe, Japan
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  • Chika Seiwa,

    1. Department of Neuro-Glia Cell Biology, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan
    2. Center for Kampo Medicine, School of Medicine, Keio University, Tokyo, Japan
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  • Kiyoto Motojima,

    Corresponding author
    1. Department of Biochemistry, Graduate School of Pharmaceutical Sciences, Meiji Pharmaceutical University, Tokyo, Japan
    • Department of Biochemistry, Graduate School of Pharmaceutical Sciences, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo 204-8588, Japan
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  • Hiroaki Asou,

    1. Department of Neuro-Glia Cell Biology, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan
    2. Center for Kampo Medicine, School of Medicine, Keio University, Tokyo, Japan
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  • Masahiro Yamamoto

    Corresponding author
    1. Department of Neuro-Glia Cell Biology, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan
    2. Tsumura Laboratories, Tsumura & Co., Ibaraki, Japan
    • Tsumura Laboratories, Tsumura & Co., 3586 Yoshiwara, Ami-machi, Inashiki-gun, Ibaraki 300-1192, Japan
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  • Requests for materials should be addressed to M. Yamamoto (hirokoma@h.email.ne.jp) or H. Asou (asou@sc.itc.keio.ac.jp)

Abstract

Recent research in neural development has highlighted the importance of markers to discriminate phenotypic alterations of neural cells at various developmental stages. We isolated a new monoclonal antibody, 4F2, which was shown to be specific for an oligodendrocyte lineage. In primary cultures of oligodendroglial and mixed neural cells, the 4F2 antibody labeled a large proportion of Sox2+, Sox10+, A2B5+, NG2+, Olig2+, O4+, and myelin basic protein (MBP)+ cells but did not label any GFAP+ or NeuN+ cells. In immunohistochemisty of rat embryos, the 4F2 antibody labeled a portion of neuroepithelial cells of the neural tube at embryonic day 9. The 4F2-positive cells were located initially in the ventricular zone as Musashi1+ Tuj1 populations and distributed throughout the striatum; thereafter, they populated the whole brain and spinal cord. These cells showed ramified processes during embryonal development. The 4F2 antigen was associated with all four isoforms of MBP in coimmunoprecipitation experiments using brain homogenates or cell lysates of cultured oligodendrocytes. Immunoscreening of a brain cDNA library identified the antigen as DEAD (Asp-Glu-Ala-Asp) box polypeptide 54 (Ddx54), a member of the DEAD box family of RNA helicases involved in RNA metabolism, transcription, and translation. Cotransfection of the Ddx54 gene with MBP isoform genes increased the nuclear localization of the 21.5-kDa MBP isoform, which has been reported to function as a nuclear signal transduction molecule. These data indicate that Ddx54 might be not only a useful marker for investigating the ontogeny of oligodendrocytes but also an important factor in oligodendrocyte differentiation and myelination. Journal of Neuroscience Research (2011) © 2011 Wiley Periodicals, Inc.

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