| Project/Area Number |
14571550
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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 |
Obstetrics and gynecology
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| Research Institution | University of Yamanashi, Interdisciplinary Graduate School of Medicine and Engineering (2003)
山梨医科大学 (2002) |
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Principal Investigator |
HOSHI Kazuhiko University of Yamanashi, Interdisciplinary Graduate School of Medicine and Engineering, Professor, 大学院・医学工学総合研究部, 教授 (20111289)
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| Co-Investigator(Kenkyū-buntansha) |
SHODA Tomoko University of Yamanashi, Interdisciplinary Graduate School of Medicine and Engineering, Research Associate, 大学院・医学工学総合研究部, 助手 (50345716)
KASAI Tsuyoshi University of Yamanashi Affiliated Hospital Dept. of Obstetric and Gynecology, Assistant Professor, 医学部附属病院, 講師 (20194699)
HIRATA Shuji University of Yamanashi, Interdisciplinary Graduate School of Medicine and Engineering, Associate Professor, 大学院・医学工学総合研究部, 助教授 (00228785)
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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 |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | sperm factor / human sperm / egg maturation / Ca oscillation / ICSI / oscilin / cDNA cloning / genomic cloning / sperm factor / PLCζ / 精巣cDNA / ヒト |
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| Research Abstract |
The aim of the research project is analysis of the sperm factor is transferred into in an egg and elicits activation of an egg. There are many unelucidated issues about the mechanism of the egg activation. We hypothesized that phospholipase C(PLC) zeta (PLCζ) which is a sperm isoform of PLC is the true sperm factor, and carried out the following studies;
(1)We screened the human cDNA library by the human PLCζ cDNA probe.
(2)We cloned two distinct cDNAs. One clone corresponed to the full length PLCζ cDNA (wild type PLCζ cDNA, and another cDNA clone lacked exon 2'-3-4 (del. 2'-3-4 variant PLCζ cDNA).
(3)To analyze of the function of the protein encoded by the wild type PLCζ cDNA and del. 2'-3-4 variant PLCζ cDNA, we synthesized and purified the wild type PLCζ and the del. 2'-3-4 variant PLCζ using prokaryotic GST-fusion protein expression system.
(4)We microinjected the recombinant proteins into a mouse egg and analyzed changes in intracellular calcium ion level using confocal laser microscope.
(5)As a result, ca oscillation was evoked in a mouse egg by microinjection of wild type PLCζ as well as del. 2'-3-4 variant PLCζ, indicating that PLCζ may be a true sperm factor.
The recombinat PLCζ synthesized in the present study might be very useful for the treatment of a certain infertility male whose sperm can not activate the egg by ICSI.
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