| Project/Area Number |
09640792
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物形態・構造
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| Research Institution | Hiroshima University |
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Principal Investigator |
NAKAMURA Masahisa HIROSHIMA UNIVERSITY, FACULTY OF SCIENCE, LABORATORY FOR AMPHIBIAN BIOLOGY, PROFESSOR, 理学部, 教授 (40130025)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Sex differentiation / SF-1 / Ad4BP / FTZ-F1 (α, β) / Sox9 (α, β) / cDNA / 5'-UTR / frog / gonad / 性分化 / FTZ-F1(α、β) / Sox9(α、β) / 5´UTR / 生殖腺 / FTZ-F1 / 性腺 / 両生類 / ノーゲン解析 / in situ ハイブリダイゼーション / 生殖細胞 / 性転換 / ゲノム構造 / FTZ-F1cDNA / Ad4BPcDNA / 5'-UTR / ツチガエル |
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| Research Abstract |
1 . SF- I /Ad4BP
We successfully cloned the SF-1/Ad4BP cDNA from the testis λgt10 cDNA library of the frog, Rana rugosa. It encoded a protein of 468 amino acids and its ORF shared 70% similarity with that of chicken OR2.1 (a SF-1/Ad4BP homologue) and 62% with bovine Ad4BP. SF-1/Ad4BP mRNA was expressed in testes, brains, kidneys and spleens , but not ovaries of adult frogs. The 5 ' -upstream region of the SF-1/Ad4BP gene including the 1st and 2nd exons was also cloned from a λ gt10 genomic DNA library using the cDNA as a probe. A sequence analysis of this gene showed that another nuclear factor ELP probably is not transcribed from the same gene.
2. FTZ-F 1
We cloned frog homologues of FTZ-F1 α and rrFTZ-F1 β cDNAs. Two types (α and β) of rrFTZ-F1 mRNAS were expressed in the embryo at stage 12. Expressions of both the α and β mRNAs became stronger in the testis of frogs at stage xxv and were the most prominent in that of frogs 2 months after metamorphosis. In addition, we cloned the regions with either exon I or II of the rrFTZ-F1 gone. Genomic structure analysis revealed that rrFTZ-F1 p is a partial exon I-truncated variant of rrFTZ-F1 α.
3. Sox9
We isolated two different mRNA isoforms ( α and β ) of the frog Sox9 gene from adult frog testis cDNAS. Sox9 expression was up-regulated in embryos after stage 16, and was seen in both developing testes and ovaries. The size of Sox9 transcripts was determined to he 7.8 knt by Northern blot analysis. In addition Sox9 α expression was found prominently in the testis and brain among various tissues of adult frogs examined, and was considerably higher than Sox9 β. The fact that Sox9 is expressed in both sexes suggests that this gene is involved in gonadal development of male and female frogs.
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