Developmental analyzes of primordial germ cells and induction of germ-line chimera in fish.

鱼类原始生殖细胞的发育分析和生殖系嵌合体的诱导。

• 批准号: 14360105
• 负责人: YAMAHA Etsuro
• 金额: $ 9.6万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14360105/
• 关键词: embryogenesis; germ-line chimera; nos 1; primordial germ cells; teleost fish; transgenic trout; vasa; GFP; nos 1; TGF-β super family; vas

1)The origin and migration route were investigated by histological section and whole mount in situ hybridization of vas mRNA at a probe in ice goby and floating goby (Gobioidei, Perciformes, Teleostei).2)Primordial germ cells were visualized at the somitogenesis stages with GFP fluorescence in herring, goldfish, loach and ice goby embryos, when fertilized eggs of these species were injected with chimeric RNAs containing GFP coding region and zebrafish nos1 3'-untranslated region.3)A technique to visualize live PGCs using chimeric RNAs containing GFP coding region and vasa 3'-untranslated region was also established.4)When visualized PGCs at the somitogenesis stage were inter-temporally transplanted into blastoderm at the blastula stage, donor PGCs moved to genital anlage of the host embryo in con-specific transplantation.5)In the case of inter-temporal, xeno-generic transplantation, for example from goldfish to loach, donor PGCs moved to the host genital anlage through the host migration route with the host PGCs. But, in some combination of xeno-generic transplantation, such as from loach to ice goby, donor PGCs could not moved to the host genital anlage.6)A technique to generate live fry from intraperitoneally transplanted primordial germ cells was established in salmonid fish.7)A technique to isolate highly pure and viable PGCs from rainbow trout by GFP-dependent flow cytometry was established.

1）通过组织学部分和整个安装式杂交在冰戈比（Ice Goby）的探头和漂浮的戈比（Gobioidei，perciformes，teleostei，teleostei）中，通过组织学部分和整个安装杂交进行了起源和迁移路线。嵌合RNA含有GFP编码区域和斑马鱼NOS1 3'-非翻译区域。3）一种使用含有GFP编码区域的嵌合RNA可视化实时PGC的技术PGCS在CON特异性移植中移至宿主胚胎的生殖器。但是，在异种生成移植的某种结合中，例如从Loach到Ice Goby，供体PGC无法移动到宿主生殖器的生殖器。6）一种技术，从腹膜内移植的原始生殖细胞中生成活肉，在鲑鱼中建立了高度纯净的pgc pgc trut trut。 已确立的。

项目成果

Yoshizaki, G.: "Visualization of primordial germ cells in transgenic rainbow trout carrying green fluorescent protein gene driven by vasa promoter"Fisheries Science. 68. 1067-1070 (2002)

Yoshizaki, G.：“携带由 vasa 启动子驱动的绿色荧光蛋白基因的转基因虹鳟鱼中原始生殖细胞的可视化”渔业科学。

Hashimoto, Y.: "Requirement of localized maternal factors for zebrafish germ cell formation."Dev.Biol.. 268. 152-161 (2004)

Hashimoto, Y.：“斑马鱼生殖细胞形成对局部母体因素的要求。”Dev.Biol.. 268. 152-161 (2004)

Kobayashi, T.: "Isolation of highly pure and viable primordial germ cells from rainbow trout by GFP-dependent flow cytometry."Mol.Reprod.Dev.. 67. 91-100 (2004)

Kobayashi, T.：“通过 GFP 依赖的流式细胞术从虹鳟鱼中分离高纯度和活性的原始生殖细胞。”Mol.Reprod.Dev.. 67. 91-100 (2004)

Yoshizaki, G., Takeuchi, Y., Kobayashi, T., Tago, Y., Takeuchi, T.: "Fish bioengineering by primordial germ cells (in Japanese)"BIO INDUSTRY. 21. 29-37 (2004)

Yoshizaki, G.、Takeuchi, Y.、Kobayashi, T.、Tago, Y.、Takeuchi, T.：“通过原始生殖细胞进行鱼类生物工程（日语）”生物工业。

Takeuchi, Y., Yoshizaki, G., Takeuchi, T.: "Generation of live fry from intraperitoneally transplanted primordial germ cells in rainbow trout."Biol.Reprod.. 69. 1142-1149 (2003)

Takeuchi, Y.、Yoshizaki, G.、Takeuchi, T.：“虹鳟鱼腹膜内移植原始生殖细胞生成活鱼苗。”Biol.Reprod.. 69. 1142-1149 (2003)