| Project/Area Number |
04671026
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | HYOGO COLLEGE OF MEDICINE |
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Principal Investigator |
KOYAMA Koji HYOGO COLLEGE OF MEDICINE, DEPARTMENT OF OBSTETRICS AND GYNECOLOGY, PROFESSOR, 医学部, 教授 (00068496)
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| Co-Investigator(Kenkyū-buntansha) |
AKATANI Akiko HYOGO COLLEGE OF MEDICINE, DEPARTMENT OF OBSTETRICS AND GYNECOLOGY, ASSISTANT PR, 医学部, 助手 (50212402)
YAMASAKI Noriyuki HYOGO COLLEGE OF MEDICINE, DEPARTMENT OF OBSTETRICS AND GYNECOLOGY, ASSISTANT PR, 医学部, 助手 (50174644)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1992: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | monoclonal antibody / fertilization / ubiquitin / human sperm / 精子先体 / ユビキチン / 精子不動化 / 精子付着精漿抗原 / 遺伝子クローニング |
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| Research Abstract |
The MAb(1G1) possessing a strong fertilization-blocking activity was prepared by immunizing BALB/c with capacitated human sperm in order to study the molecular nature of sperm antigens relevant to fertilization. The MAb 1G1 strongly inhibited human sperm penetration into zona-free hamster eggs. It reacted to the apical portion of acrosome-reacted human sperm, but did not react to the live sperm before acrosome reaction, when the immunofluorescence staining were performed. In paraffin-embedded testis sections, the round spermatids, spermatocytes and spermatozoa were stained with the MAb 1G1. The spermatogonia, Sertoli cells, Leydig cells, and other somatic tissues were not stained. The sperm of monkeys, bulls, mice and boars were not stained. Therefore, the staining of sperm was species specific. The antigen corresponding to the MAb 1G1 showed a band at a 27kDa by immunoblotting. The reactivity of the antigenic components were not destroyed by periodic acid treatment. As a result, it was postulated that the antigenic molecule might be a peptide. After screening the human testis cDNA library with the MAb 1G1, we isolated a cDNA clone encoding the antigen. According to the nucleotide sequences of th cDNA clone, they showed that the cDNA contained human poly-ubiquitin DNA.These results suggests that ubiquitin might play an important role in human fertilization.
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