| Project/Area Number |
15590490
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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 |
Laboratory medicine
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| Research Institution | The University of Tokushima |
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Principal Investigator |
UMENO Mayumi The University of Tokushima, School of Medicine, Assistant, 医学部, 助手 (00213497)
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| Co-Investigator(Kenkyū-buntansha) |
SHINKA Toshikatsu The University of Tokushima, Graduate School, Institute of Health Biosciences, Assistant Professor, 大学院・ヘルスバイオサイエンス研究部, 助教授 (10311820)
NAKAHORI Yutaka The University of Tokushima, Graduate School, Institute of Health Biosciences, Professor, 大学院・ヘルスバイオサイエンス研究部, 教授 (10172389)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Y chromosome deletions / male infertility / AZF / multiplex PCR / microchip electrophoresis / HSFY |
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| Research Abstract |
1.A rapid and simple system of detecting deletions on the Y chromosome related with male infertility using multiplex PCR
Around 10% of males with idiopathic azoospermia or oligozoospermia, which are important causes of male infertility, have partial deletions on the long arm of the Y chromosome. To develop a rapid and accurate detection system for screening major Y deletions found in infertile men, we developed a multiplex PCR system that can simultaneously amplify five loci on the Y chromosome, SRY,AMELY,DBY,RBMY,DAZ and one locus on the X chromosome, AMELX. The size of the PCR products was designed to increase gradually from the distal Yp to the distal Yq. Our system could detect deletions of three major candidate regions for the azoospermic factor, AZFa, AZFb and AZFc on the Y chromosome together with sex. Moreover, the multiplex PCR system was combined with microchip-based electrophoresis, and the PCR products derived from each locus were separated within 4min. Our system is useful for screening Y chromosomes bearing the structural anomalies including three major AZF deletions found among azoospermic or oligozoospermic males.
2.Study of genetic background in male infertility
We focused on HSFY, which is mapped on AZFb. Expression analysis of HSFY and mHSFYL unveiled that they are expressed predominantly in testis. Furthermore, immunnohistochemistry of HSFY in testis showed that its expression is restricted to both Sertoli cells and spermatogenic cells, and that it exhibits a stage-dependent translocation from the cytoplasm to the nucleus in spermatogenetic cells during spermatogenesis. These results may suggest that deletion of HSFY is involved in azoospermia or oligospermia.
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