| Project/Area Number |
10470350
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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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 | Hyogo college of Medicine |
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Principal Investigator |
KOYAMA Koji Hyogo College of Medicine, Department of Obstetrics and Gynecology,, 医学部, 教授 (00068496)
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| Co-Investigator(Kenkyū-buntansha) |
KOMORI Shinji Hyogo College of Medicine, Department of Obstetrics and Gynecology,, 医学部, 講師 (60195865)
AKATANI-HASEGAWA Akiko Hyogo College of Medicine, Institute for Advanced Medical Sciences, Laboratory of Developmental Biology and Reproduction, 医学部, 助手 (50212402)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥12,200,000 (Direct Cost: ¥12,200,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1998: ¥9,100,000 (Direct Cost: ¥9,100,000)
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| Keywords | Zona pellucida / Pig ZPA / Transgenic mice / Male infertility / Sperm function diagnosis / In vitro fertilization / ZPA / 受精 / 受精能獲得 / 精子レセプター / 先体反応 / 不妊症 |
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| Research Abstract |
The objective of this study is to produce zona pellucida for use in human sperm function diagnosis. We attempted to make transgenic mice expressing pig ZPA protein to which human sperm binds. The reason why we selected pig ZPA instead of human is that another group had already started a similar study using the human ZPA gene. The transgene was constructed with the promoter region of mouse ZPA (1.4kb) and the minigene including pig ZPA full-length DNA (6.8kb) and polyadenylation signal of rabbit β-globin (0.5kb). The promoter DNA fragment of mouse ZPA was kindly provided by Dr.J.Dean of NIH inUA.In order to introduce the transgene into mouse embryos BDF1 female mice were treated with PMS/hCG for superovulation before mating. The injected embryos was implanted in the oviducts of pseudo-pregnant mice. The incorporation of the transgene were examined by PCR analysis using DNA extracted from the tails. The transgene was detected in four newly born mice. Two (_<2, >8) of them transferred the transgene genes to their progenies. The corresponding transcripts were detected not only in ovarian tissue but also in other tissues including liver, heart, lung. But protein expression was only observed in oocytes and zona pellucida. The heterologous sperm binding capacity of the zona pellucida from the transgenic mice is currently being investigated.
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