| Co-Investigator(Kenkyū-buntansha) |
NOMURA Takako Okayama Univ., Graduate School of Medicine and Dentistry. Dept.of Cytology and Histology, Assistant, 大学院・医歯学総合研究科, 助手 (20116437)
SASAKI Junzo Okayama Univ., Graduate School of Medicine and Dentistry. Dept.of Cytology and Histology, Professor, 大学院・医歯学総合研究科, 教授 (30093686)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
【Introduction】The mechanism of ovulation has not been clarified yet. To study ovulatory mechanism, genetic dynamics during the time before and after ovulation was analyzed with DNA micro-array and observed after immunohistochemical stains were given to the noticeable genes to see when and in which cells in the oocytes, up and down regulation occurred.
【Materials and Methods】Experiments were carried out in 2 groups of Wistar rats. In the 1^<st> group, oocytes were treated by various immunohistochemical stains 0 to 42 days after birth to observe the changes in apoptosis, cell cycle, sexual hormones, and vascular endothelia in the ovary. In the 2^<nd> group, rats without any treatment 26 days after birth were used as a control. Pregnant mere's gonadotropin(PMSG) was administered to rats 24 days after birth. After the PMSG administration, human chorionic gonadotropin(hCG) was further administered to them. Ten hours after the administration, the ovary of each of these rats was extirpated und
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er pentobarbital anesthesia and mRNA was biochemically extracted from the ovary to analyze the genetic changes with DNA micro-array. Oval tissues of each of the control, PMSG-administered, and PMSG/hCG-administered rats was observed after immunohistochemical stains as done in group 1.
【Results】In the 1^<st> group, it was found that the ova were eliminated by apoptosis twice : during the period from 0 to 3 days after birth and during that from 25 to 30 days after birth. In the 2^<nd> group, it was demonstrated that all oocytes became in the state of proliferation as PMSG administration inhibited apoptosis. By hCG administration, cytochrome P-450 arom was decreased while prostaglandine endoperoxide synthase, blood coagulation systems, protease groups, various groups of cytokine, and S 100 calcium binding proteins were increased.
【Discussion and conclusion】As a result of DNA micro-array analyses, lots of varied genes such as cytochrome P-450, blood coagulation systems, proteases etc. were the same as the factors which is now reportedly related with the time of ovulation. In this study 2 points were clarified : firstly, the ova reached the fixed sexual cycle after being selected twice during the immature period and secondly, various S 100 calcium binding proteins greatly changed immediately before ovulation. However, dynamic mechanism of S 100 proteins in the ovary still remains unclear and further study on this issue is needed. Less
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