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: 12460081
• 负责人: YAMAHA Etsuro
• 金额: $ 9.22万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460081/
• 关键词: embryogenesis; germ-line chimera; goldfish; primordial germ cells; rainbow trout; transgenic trout; vasa; vas-GFP; 生殖系列キメラ; PGCs; 中胚葉誘導; vasa; vasa-GFP; 生殖細胞キメラ

1) Distribution of vas mRNA were analyzed by whole mount in situ hybridization during embryogenesis in goldfish. There are some difference of distribution of vas mRNA between goldfish and zebrafish.2) In goldfish, functional PGCs were differentiated from mesoderm deficient embryo.3) In goldfish, PGCs migrated passively by convergence and extension movement during gastrulation.4) Germ-line chimera was successfully induced by transplantation of graft, containing PGCs, to host embryo at the blastula stage.5) Sperm derived from donor was only generated in germ-line chimera by using sterile hybrid as host embryo.6) Transgenic trout strains having GFP-labeled PGCs were established.7) A technique to purify live PGCs using a flow cytometer was developed.8) Genes expressed specifically in PGCs were isolated by the cDNA subtraction method.9) A technique for PGCs transplantation using isolated GFP-labeled PGCs was developed in order to establish a procedure for re-introducing donor PGCs in to the germline to recipient fish.

1）通过在金鱼中胚胎发生过程中，通过整个植物原位杂交分析了VAS mRNA的分布。金鱼和斑马鱼之间的VAS mRNA分布有所不同。2）在金鱼中，功能性PGC与中胚层不足的胚胎区分开。3）在金鱼中，PGCS在胃部期间通过胃部的杂物连续诱导的embrantation intplula intplula to and and to and off pgc，pGC通过胃部的胃线在胃部中被动地迁移和延伸。仅通过使用无菌杂交作为宿主胚胎才能在种系嵌合中产生供体。6）建立具有GFP标记PGC的转基因鳟鱼菌株。7）7）一种使用流式细胞仪的基因纯化PGC的基因，该基因在PGC中脱离了CDNNA suplique.9）开发了GFP标记的PGC，以建立一种将供体PGC重新引入到受体鱼类的生殖线的程序。