Summary of Research Project Results under JSPS FY2001
"Research for the Future Program"

1.Research Institution Tokyo University of Fisheries
2.Research Area Life Sciences
3.Research Field Molecular Bioengineering of Food Animal Protein Resources
4.Term of Project FY 1997 〜 FY 2001
5.Project Number 97L00902
6.Title of Project Genetic control of the Reproduction in Aquatic Animals of Important Aquaculture Resources

7.Project Leader
Name Institution,Department Title of Position
Takashi Aoki Tokyo University of Fisheries, Grduate School of Fisheries Professor

8.Core Members

Names Institution,Department Title of Position
Seiichi Watanabe Tokyo University of Fisheries, Faculty of Fisheries Professor
Nobuaki Okamoto Tokyo University of Fisheries, Faculty of Fisheries Professor
Nobuhiko Taniguchi Tohoku University, Graduate School of Agriculture Professor

9.Summary of Research Results

1. We have isolated and cloned many putative biodefense and immuno response-related genes from the Japanese flounder, Rainbow trout, red seabream, and carp; these include cytokines, cytokine receptors, cell surface molecules, transcription factors, antimicrobial proteins, proteases, and protease inhibitors following EST analyses.
2. We have developed a microarray of 800 different genes of Japanese flounder. We confirmed that this microarray was useful for characterization of gene expression patterns of several cells and organs of Japanese flounder.
3. Heat shock protein (HSP) over expressed transgenic zebrafish embryos showed extensive apoptosis and abnormal morphogenesis. The enhanced HSP70 levels regulated bone morphogenetic protein 4 (BMP4) mediated apoptotic signaling in early development.
4. MAP-kinase and taurine transporter genes were cloned from fish cells and demonstrated to be as osmo-responsive genes for candidates to produce osmo-resistant fish.
5. We mapped quantitative trat loci (QTLs) for IPN resistance in a segregated population of backcross fish, derived from out-crossing the resistant strain of rainbow trout, RT-201 and the susceptible one, RT-101.
6. Foreign gene transfer method were established on red sea bream using sperm.The expression vectors using fish virus promoter were constructed and confirmed.
7. Cloned red sea bream, Pagrus major was produced by chromosome manipulation, clonal status of the fish was confirmed by multi-locus DNA fingerprinting. Sex differentiation and sex control of the fish were also clarified.
8. We aim to use primordial germ cells (PGCs) as an initial material of such a stem cell line in fish. As a first step in establishing a fish cell line derived from PGCs, we have developed techniques for visualization, isolation, and transplantation of viable fish PGCs.
9. DNA markers to evaluate genetic divergence were developed in order to verify their efficiency in the population genetic analysis for broodstock management in stock enhancement and aquaculture avtivities.
10. We investigated the genetic impact of stock-enhancement programs using simulation models, and constructed an effective strategy for this program. The research focuses on the conservation and the sustainable use of genetic resource in the future.

10.Key Words

(1)Diseases resistance、(2)Immune- and defense-related genes、(3)stress response
(4)Transgenic fish、(5)quantitative trait loci、(6)genetic polymorphism
(7)broodstock management、(8)simulation models、(9)cloned fish