title: Generation of germ-line chimera zebrafish using primordial germ cells isolated from cultured blastomeres and cryopreserved embryoids.
abstract: Primordial germ cells (PGCs) are the only cells in developing embryos with the potential to transmit genetic information to the next generation. In our previous study, a single PGC transplanted into a host differentiated into fertile gametes and produced germ-line chimeras of cyprinid fish, including zebrafish. In this study, we aimed to induce germ-line chimeras by transplanting donor PGCs from various sources (normal embryos at different stages, dissociated blastomeres, embryoids, or embryoids cryopreserved by vitrification) into host blastulae, and compare the migration rates of the PGCs towards the gonadal ridge. Isolated, cultured blastomeres not subject to mesodermal induction were able to differentiate into PGCs that retained their motility. Moreover, these PGCs successfully migrated towards the gonadal ridge of the host and formed viable gametes. Motility depended on developmental stage and culture duration: PGCs obtained at earlier developmental stages and with shorter cultivation periods showed an increased rate of migration to the gonadal ridge. Offspring were obtained from natural spawning between normal females and chimeric males. These results provide the basis for new methods of gene preservation in zebrafish.
authors.affiliation: Nanae Fresh Water Laboratory, Field Science Center of Northern Biosphere, Hokkaido University, Nanae, Japan. yutakakjp@hokudai-nanae.jp
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kawakami
authors.forename: Rie
authors.initials: R
authors.lastname: Goto-Kazeto
authors.forename: Taiju
authors.initials: T
authors.lastname: Saito
authors.forename: Takafumi
authors.initials: T
authors.lastname: Fujimoto
authors.forename: Shogo
authors.initials: S
authors.lastname: Higaki
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Arai
authors.forename: Etsuro
authors.initials: E
authors.lastname: Yamaha
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001757:Blastomeres / Q000166:cytology* / Q000637:transplantation; D036703:Blastula / Q000166:cytology / Q000196:embryology / Q000637:transplantation; D002465:Cell Movement; D002478:Cells, Cultured / Q000166:cytology; D002678:Chimera; D015925:Cryopreservation; D004625:Embryo, Nonmammalian / Q000166:cytology; D058732:Embryoid Bodies / Q000166:cytology* / Q000637:transplantation*; D005854:Germ Cells / Q000166:cytology* / Q000637:transplantation*; D006066:Gonads; D049452:Green Fluorescent Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D013194:Staining and Labeling; D014180:Transplantation; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029961:Zebrafish Proteins; C440015:nanos3 protein, zebrafish; D049452:Green Fluorescent Proteins