Chicken Dead End Homologue Protein is a Nucleoprotein of Germ Cells Including Primordial Germ Cells
Chicken Dead End Homologue Protein is a Nucleoprotein of Germ Cells Including Primordial Germ Cells
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DOI：
10.1262/jrd.20154
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发表时间：
2009-04-01
影响因子：
1.8
通讯作者：
Hattori, Masa-aki
中科院分区：
文献类型：
--
作者：
Aramaki, Shinya;Kubota, Kaiyu;Hattori, Masa-aki
编码RNA结合蛋白的死端基因主要在脊椎动物的生殖细胞中表达。最近，我们克隆了鸡死端同源物（CDH），并表明CDH mRNA的表达是高度特异性的原始生殖细胞（PGCs）在早期胚胎阶段。迄今为止，由于缺乏抗体，死端蛋白在生殖细胞中的亚细胞定位在很大程度上是未知的。本研究制备了鸡死端同源物（CDH）的多克隆抗体，以阐明CDH在生殖细胞中的亚细胞定位。为了与CDH进行比较研究，还提出了一种抗鸡血管同源物（CVH）（一种众所周知的生殖细胞标记物）的多克隆抗体。CDH蛋白的免疫印迹分析显示，在卵巢和睾丸蛋白中存在分子大小约为60 kDa的单一条带。免疫荧光研究表明，CDH蛋白只定位于原始生殖细胞（PGCs）和生殖细胞在后期阶段的细胞核，而CVH定位于细胞质。有趣的是，分布在生精上皮基底侧的生殖细胞，如精原细胞，CDH蛋白呈强阳性。目前的研究提供了新的证据，CDH是一个核蛋白的生殖细胞，包括PGCs。
The dead end gene, coding an RNA binding protein, is predominantly expressed in the germ cells of vertebrates. Recently, we cloned chicken dead end homologue (CDH) and showed that expression of CDH mRNA is highly specific to primordial germ cells (PGCs) at early embryonic stages. To date, the subcelluler localization of Dead end protein in germ cell has been largely unknown due to lack of an antibody. Here, we raised a polyclonal antibody against chicken dead end homologue (CDH) to elucidate its subcellular localization in the germ cells. For comparative studies with CDH, a polyclonal antibody against chicken vasa homologue (CVH), a well-known germ cell marker, was also raised. Immunoblotting analysis for CDH protein showed a single band with a molecular size of approximately 60 kDa in the ovarian and testicular proteins. Immunofluorescence studies revealed that CDH protein was exclusively localized in the nuclei of primordial germ cells (PGCs) and germ cells at later stages, while CVH was localized in the cytoplasm. Interestingly, the germ cells distributed at the basal sides of seminiferous epithelia, such as spermatogonia, were strongly positive to CDH protein. The current study provides novel evidence that CDH is a nucleoprotein of germ cells, including PGCs.