Investigation for Common Molecular Mechanisms by Comparison of EST from Purified Germ Cells

通过比较纯化生殖细胞的 EST 研究常见分子机制

• 批准号: 15310133
• 负责人: TANAKA Minoru
• 金额: $ 8.83万
• 依托单位: National Institute for Basic Biology (2004)Okazaki National Research Institutes (2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15310133/
• 关键词: germline; EST; development; differentiation; vertebrates; 生殖細胞; 網羅的解析; メダカ; 生殖腺形成; 性決定分化

medaka embryos are transparent, which allows us to monitor the development process occuring in morphogenesis in vivo. We have developed transgenic medaka using the regulatory regions of germline-specific expressed gene, vasa, fused with the green fluorescent gene. With this transgenics, we can observe the development of germline, the cell lineage for the future eggs or sperms. We have established the skills for isolating several germ cells and generated cDNA libraries for ESTs expressed exclusively in germline.Our initial purpose was to compare the medaka ESTs with mouse germline ESTs to reveal the genes commonly expressed beyond the species. We could not generate the libraries good enough for representing medaka germline ESTs. However, among the thousnads of ESTs we sequenced, several interesting ESTs have been found. By the expression and functional analyses of the ESTs, we successfully kept track of germline from fertilized eggs. Medaka germline is detected at the gastrulation stage, which has been suggesting that development of germline is similar to that seen in mammals. Our results indicate that germline development is more likely to that of other vertebrates where germline can be marked by specific cellular structures. Furthermore we demonstrated thatgermline goes through three steps of developmental process with three different modes of migration towards gonadal area.

Medaka胚胎是透明的，这使我们能够监测体内形态发生中发生的发展过程。我们使用种系特异性表达基因VASA的调节区域开发了转基因MEDAKA，该基因与绿色荧光基因融合在一起。有了这种转基因，我们可以观察到生殖线的发展，即未来卵或精子的细胞谱系。我们已经建立了隔离几个生殖细胞的技能，并生成了仅在种系中表达的EST的cDNA文库。我们的最初目的是将Medaka EST与小鼠种系EST进行比较，以揭示通常表达的基因以外的物种。我们无法产生足够好的图书馆来代表Medaka种系EST。但是，在我们测序的EST的Thousnads中，发现了一些有趣的EST。通过EST的表达和功能分析，我们成功地跟踪了受精卵的种系。在胃阶段检测到Medaka种系，这表明种系的发育与哺乳动物中的生殖线相似。我们的结果表明，种系的发育更有可能由特定细胞结构标记的其他脊椎动物的发育。此外，我们证明了格林线经历了三个不同的发展过程，并向性腺区域进行了三种不同的迁移方式。

项目成果

Neurogenin3 delineates the earliest stages of spermatogenesis in the mouse testis

• DOI: 10.1016/j.ydbio.2004.01.036
• 发表时间: 2004-05-13
• 期刊: DEVELOPMENTAL BIOLOGY
• 影响因子: 2.7
• 作者: Yoshida, S;Takakura, A;Nabeshima, Y
• 通讯作者: Nabeshima, Y

Expression of aromatase mRNA and effects of aromatase inhibitor duing ovarian development in the medaka, Oryzias latipes.

青鳉卵巢发育过程中芳香酶 mRNA 的表达及芳香酶抑制剂的影响。

• 发表时间: 2004
• 期刊: Journal of Experimental Zoology 301
• 作者: T.Morii;et al.;Aya Suzuki
• 通讯作者: Aya Suzuki

Ara, et al.: "Impaired colonization of the gonads by primordial germ cells in mice lacking a chemokine, stromal cell-derived factor-1 (SDF-1)."Proceedings of the National Academy of Science, USA. 100. 5319-5323 (2003)

Ara 等人：“缺乏趋化因子基质细胞衍生因子 1 (SDF-1) 的小鼠中原始生殖细胞对性腺的定植受损。”美国国家科学院院刊。

Morinaga, et al.: "Mutations affecting gonadal development in Medaka, Oryzias latipes."Mechanism of Development. in press. (2004)

Morinaga 等人：“影响青鳉、Oryzias latipes 性腺发育的突变。”发育机制。

Naruse, et al.: "A Medaka Gene Map : The Trace of Ancestral Vertebrate Proto-chromosomes Revealed by Comparative Gene Mapping"Genome Research. in press. (2004)

Naruse 等人：“青鳉基因图谱：比较基因图谱揭示的祖先脊椎动物原染色体的踪迹”基因组研究。