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This Article Full Text Full Text (PDF) All Versions of this Article: 78/1/159    most recent biolreprod.107.060038v2 biolreprod.107.060038v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Saito, T. Articles by Yamaha, E. Search for Related Content PubMed PubMed Citation Articles by Saito, T. Articles by Yamaha, E. Agricola Articles by Saito, T. Articles by Yamaha, E.

Xenogenesis in Teleost Fish Through Generation of Germ-Line Chimeras by Single Primordial Germ Cell Transplantation¹

Nanae Fresh Water Laboratory,³ Field Science Center for Northern Biosphere, Hokkaido University, Hokkaido 041-1105, Japan Laboratory of Aquaculture, Genetics and Genomics,⁴ Faculty of Fisheries Sciences, Hokkaido University, Hokkaido 041-8611, Japan

ABSTRACT

Primordial germ cells (PGCs) are the only cells in developing embryos with the potential to transmit genetic information to the next generation. PGCs therefore have the potential to be of value for gene banking and cryopreservation, particularly via the production of donor gametes with germ-line chimeras. Currently, it is not clear how many PGCs are required for germ-line differentiation and formation of gonadal structures. In the present study, we achieved complete germ-line replacement between two related teleost species, the pearl danio (Danio albolineatus) and the zebrafish (Danio rerio), with transplantation of a single PGC into each host embryo. We isolated and transplanted a single PGC into each blastula-stage, zebrafish embryo. Development of host germ-line cells was prevented by an antisense dead end morpholino oligonucleotide. In many host embryos, the transplanted donor PGC successfully migrated toward the gonadal anlage without undergoing cell division. At the gonadal anlage, the PGC differentiated to form one normally sized gonad rather than the pair of gonads usually present. Offspring were obtained from natural spawning of these chimeras. Analyses of morphology and DNA showed that the offspring were of donor origin. We extended our study to confirm that transplanted single PGCs of goldfish (Carassius auratus) and loach (Misgurnus anguillicaudatus) can similarly differentiate into sperm in zebrafish host embryos. Our results show that xenogenesis is realistic and practical across species, genus, and family barriers and can be achieved by the transplantation of a single PGC from a donor species.

developmental biology, early development, embryo, gametogenesis, primordial germ cells

¹Supported by the 21st Century COE program for Graduate School of Fisheries Sciences of Hokkaido University from MEXT of the Japanese government and a grant-in-aid for Young Scientists (B) from JSPS to T.S. (19780412) and R.G.-K. (18780140).

Correspondence: ²Taiju Saito, Nanae Fresh Water Laboratory, Field Science Center for Northern Biosphere, Hokkaido University, Sakura, Nanae, Kameda, Hokkaido 041-1105, Japan. FAX: 81 138 65 2239; e-mail: taiju{at}fish.hokudai.ac.jp