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| Content Provider | Springer Nature Link |
|---|---|
| Author | Yoshizaki, G. Takeuchi, Y. Kobayashi, T. Takeuchi, T. |
| Copyright Year | 2003 |
| Abstract | If in vitro-cultured cells could be converted into individual fish, they would have numerous applications in the field of genome biology and biotechnology. In spite of the benefits, however, no such system is presently available. Because primordial germ cells (PGCs) have the potential to be converted into individual fish via maturation and fertilization processes, we chose them to be the starting material for our system. As the first step, we visualized live PGCs in rainbow trout. Because the vasa transcript is restricted to the germ cell lineage, its regulatory regions are activated only in PGCs. Therefore, we produced transgenic strains carrying the green fluorescent protein (GFP) gene driven by the vasa gene regulatory regions. The resulting transgenic embryos showed green fluorescence specifically in PGCs. As the second step, the GFP-labeled PGCs were purified by flow cytometry. The genital ridges isolated from the transgenic embryos were dissociated by trypsin and sorted into GFP-positive and GFP-negative cells. The GFP-positive cells possessed morphological characteristics typical of PGCs. In addition, the vasa gene was expressed only in the GFP-positive cells, confirming that they were PGCs. To obtain functional gametes derived from isolated PGCs, a technique for converting PGCs into eggs and sperm is necessary. For this purpose, we developed a method for transplanting PGCs into developing embryos in order to incorporate them into the germ cell lineage of the recipient embryos. Approximately 10 PGCs isolated from newly hatched embryos were transplanted into the peritoneal cavity of recipient hatchlings. The transplanted PGCs actively migrated towards, and were finally incorporated into, the genital ridge. The PGCs that settled in the genital ridges of the recipient embryos proliferated, started meiosis, and differentiated into eggs and sperm in synchrony with the germ cells of allogenic recipient embryos. Further, the donor-derived gametes produced normal progenies through fertilization. These techniques, in combination with in vitro culture, genetic modification, and cryopreservation of PGCs, are expected to have numerous applications in the field of fish bioengineering. |
| Starting Page | 453 |
| Ending Page | 457 |
| Page Count | 5 |
| File Format | |
| ISSN | 09201742 |
| Journal | Fish Physiology and Biochemistry |
| Volume Number | 28 |
| Issue Number | 1-4 |
| e-ISSN | 15735168 |
| Language | English |
| Publisher | Kluwer Academic Publishers |
| Publisher Date | 2003-01-01 |
| Publisher Place | Dordrecht |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Animal Biochemistry Hydrobiology Zoology Animal Anatomy / Morphology / Histology Animal Physiology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Physiology Medicine Biochemistry Aquatic Science |
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