卵胞発育・排卵における細胞周期調節因子の役割と病因的意義

细胞周期调节因子在卵泡发育和排卵中的作用和病理意义

• 批准号: 12770932
• 负责人: 山本 百合恵
• 金额: $ 1.28万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12770932/
• 关键词: 顆粒膜細胞; 免疫染色; 卵巣; 排卵誘発; FSH; フローサイトメトリー

3週齢の幼若ラットにPregnant Mare Serum Gonadotropin (PMSG) 5IUの腹腔内注射を行い,その46時間後にhuman Chorionic Gonadotropin (hCG) 5IUの腹腔内注射を行った。PMSG投与前,投与後,hCG投与後の各段階でラットから卵巣を摘出,ホルマリン固定の上切片を作製し,PCNA,cyclinD2,cyclinE,p27,estradiol,progesteroneの各抗体による免疫染色を行った。A)卵胞内でPMSG投与後にestradiolの発現が増強し,hCG投与後24時間で減弱した。一方,hCG投与後36時間でprogesteroneの発現が増強した。B)卵胞内のPCNAの発現は,PMSG投与前よりPMSG投与後で増強し,hCG投与後に減弱した。C)cyclinD2,cyclinEの発現は,PMSG投与前よりPMSG投与後で増強し,hCG投与後progesteroneの発現と一致して減弱した。D)p27の発現は,hCG投与後progesteroneの発現と一致して減弱した。以上より,卵胞内での顆粒膜細胞の増殖・分化に細胞周期調節因子が関与していることが示唆された。また,その発現調節にPMSGやhCGの上位中枢からの性腺刺激ホルモンおよびestradiolやprogesteroneの性腺ホルモンが関与している可能性が示唆された。

Pregnant Mare Serum Gonadotropin (PMSG) 5IU intraperitoneal injection at 3 weeks and human Chorionic Gonadotropin (hCG) 5IU intraperitoneal injection at 46 hours. Before and after PMSG administration,hCG administration and after administration, the eggs were extracted and fixed on the upper sections of the cells. Immunostaining was performed for PCNA, cyclin D2, cyclin E,p27,estradiol and progesterone. A) The expression of estradiol increased after PMSG administration and decreased 24 hours after hCG administration. On the other hand, 36 hours after hCG administration, the occurrence of progesterone increased. B) PCNA expression in oocytes increased before PMSG administration and after PMSG administration, but decreased after hCG administration. C) CyclinD2,cyclinE expression was decreased,PMSG expression was increased,hCG expression was decreased. D)p27 expression was decreased,hCG expression was consistent with the expression of progesterone. The above results indicate that cell cycle regulators are involved in the proliferation and differentiation of granular membrane cells in oocytes. Therefore, the possibility of gonadal stimulation of PMSG and hCG in the upper center of the brain and the gonadal stimulation of estrol and progesterone in the middle of the brain is indicated.