A new monoclonal antibody, 4F2, specific for the oligodendroglial cell lineage, recognizes ATP-dependent RNA helicase Ddx54 : possible association with myelin basic protein
A new monoclonal antibody, 4F2, specific for the oligodendroglial cell lineage, recognizes ATP-dependent RNA helicase Ddx54 : possible association with myelin basic protein
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一种新的单克隆抗体 4F2，对少突胶质细胞谱系具有特异性，可识别 ATP 依赖性 RNA 解旋酶 Ddx54：可能与髓磷脂碱性蛋白相关
DOI：
10.1002/jnr.22736
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Yamamoto M
中科院分区：
文献类型：
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作者：
Ueki T;Tsuruo Y;Yamamoto Y;Yoshimura K;Takanaga H;Seiwa C;Motojima K;Asou H;Yamamoto M
近年来在神经发育方面的研究强调了标记物对神经细胞在不同发育阶段的表型改变的重要性。我们分离了一种新的单克隆抗体，4F2，它被证明是特异性的少突胶质细胞谱系。在少突胶质细胞和混合神经细胞的原代培养中，4F2抗体标记了大部分Sox2+、Sox10+、A2B5+、NG2+、Olig2+、O4+和髓鞘碱性蛋白(MBP)+细胞，但未标记任何GFAP+或NeuN+细胞。在胚胎免疫组织化学中，4F2抗体标记了胚胎第9天的部分神经管神经上皮细胞。4F2‐阳性细胞最初以Musashi1+Tuj1−群的形式分布在心室区，然后分布在纹状体中；此后，它们填充了整个大脑和脊髓。这些细胞在胚胎发育过程中表现出分枝过程。在使用脑匀浆或培养少突胶质细胞的细胞裂解物进行的共免疫沉淀实验中，4F2抗原与MBP的所有四种亚型相关。脑cDNA文库的免疫筛选鉴定该抗原为DEAD (Asp‐Glu‐Ala‐Asp) box多肽54 (Ddx54)，是参与RNA代谢、转录和翻译的RNA解旋酶DEAD box家族的一员。Ddx54基因与MBP异构体基因的共转染增加了21.5 - kDa MBP异构体的核定位，该异构体已被报道为核信号转导分子。这些数据表明，Ddx54不仅可能是研究少突胶质细胞个体发生的有用标记物，而且可能是少突胶质细胞分化和髓鞘形成的重要因素。神经科学研究杂志（2011）©2011 Wiley期刊，Inc。
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.