| Project/Area Number |
14560231
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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 |
Applied animal science
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| Research Institution | IBARAKI UNIVERSITY |
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Principal Investigator |
KANAZAWA Takuya Ibaraki Univ., Coll. of Agric., Assist.Prof., D.V.M., Ph.D., 農学部, 助手 (70272119)
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| Co-Investigator(Kenkyū-buntansha) |
OGAWA Yasuki Ibaraki Univ., Coll. of Agric., Assoc.Prof., D.V.M., Ph.D., 農学部, 助教授 (00302331)
GOTOH Hideo Natl.Inst.Agrobiol.Sci., Head, Ph.D.D.V.M., 生体防御研究グループ, チーム長 (70211919)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | spermatogonia / monoclonal antibody / differential display / mammals / testis / 雄性生殖細胞 / 哺乳動物 / 蛍光抗体組織染色法 |
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| Research Abstract |
This project was conducted to explore genes specifically expressed during postnatal resumption of growth and differentiation of spermatogonial cells, and to characterize monoclonal antibodies that specifically identify cells in seminiferous tubules of the mammals. First, to characterize tissue architecture of the goat testis, immunofluorescence staining procedure using commercially available antibodies against type IV collagen or vimentin was established : This procedure successfully revealed the basement membrane of the seminiferors tubules and somatic cells in the goat testis. Next, double immunofluorescence staining using 11 types of monoclonal antibodies, which had previously been produced against goat testicular cells, and commercial anti-collagen and vimentin antibodies in combination, characterized Leydig cells, Sertoli cells, spermatogonia (gonocytes), spermatocytes and spermatids (acrosome and flagella) in the specimen of the goat testis. The three of these 11 monoclonal antibodies has also been shown to identify Leydig cells, Sertoli cells and spermatocytes of the mouse. Third, genes that are specifically expressed during the resumption stage of spermatogenesis, were explored by differential display analysis of mRNA isolated from either day 8 or day 14 of neonatal mouse testis. More than several cDNA fragments detected, and cDNA fragments were isolated and incorporated into TA cloning vector for cloning. Cloned cDNAs are now either sequenced for homology analysis, labeled with digoxgenin used for hybridization probes, and 5'RACE to isolate complete length of cDNA.
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