title: A new monoclonal antibody, 4F2, specific for the oligodendroglial cell lineage, recognizes ATP-dependent RNA helicase Ddx54: possible association with myelin basic protein.
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.
authors.affiliation: Department of Neuro-Glia Cell Biology, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Ueki
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Tsuruo
authors.forename: Yuta
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Kazunori
authors.initials: K
authors.lastname: Yoshimura
authors.forename: Hiromi
authors.initials: H
authors.lastname: Takanaga
authors.forename: Chika
authors.initials: C
authors.lastname: Seiwa
authors.forename: Kiyoto
authors.initials: K
authors.lastname: Motojima
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Asou
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yamamoto
mesh_terms: D000367:Age Factors; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000943:Antigens, Differentiation / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D053487:DEAD-box RNA Helicases / Q000276:immunology* / Q000378:metabolism; D004622:Embryo, Mammalian; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D047468:Immunoprecipitation; D008297:Male; D004676:Myelin Basic Protein / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D009836:Oligodendroglia / Q000378:metabolism*; D011247:Pregnancy; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D004676:Myelin Basic Protein; D020033:Protein Isoforms; C036505:oligodendrocyte O antigen, mouse; C506751:Ddx5 protein, mouse; D053487:DEAD-box RNA Helicases