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

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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.