MOLECULAR MECHANISMS OF TCDD INDUCED ANOVULATION

TCDD 诱导无排卵的分子机制

• 批准号: 2018926
• 负责人: Ok-Kyong Park-Sarge
• 金额: $ 7.35万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2018926
• 关键词: aromatic hydrocarbon receptor; dioxins; environmental toxicology; estrogen receptors; gel mobility shift assay; genetic promoter element; genetic transcription; in situ hybridization; laboratory rat; molecular pathology; ovulation; progesterone receptors; receptor binding; receptor expression; western blottings

DESCRIPTION: This proposal is designed to test the hypothesis that TCDD inhibits ovulation by interruption of estrogen receptor (ER) and/or progesterone receptor (PR) mediated transcriptional activation of target genes in ovarian follicles. One specific aim is to determine cellular targets for TCDD action in the ovary by localizing aryl hydrocarbon receptors (Ah-Rs) and Ah-R nuclear translocators (ARNTs), utilizing in situ hybridizations in immature rat ovaries at various stages of follicular development and ovulation. The second specific aim is to determine whether and at what level TCDD alters the ER and/or PR mediated signaling pathways in the ovary. The ER and PR expression in the ovary will be analyzed by mRNA analysis by in situ hybridization or RT-PCR and through protein analysis using Western blots from immature gonadotropin treated rat ovaries. The ability of TCDD to inhibit estrogen or progesterone stimulated target gene transcription will be studied by examining the binding ability of Ers and Prs to DNA (gel shift analysis) and the ability to transcribe the promoter of target genes.

描述：本提案旨在检验TCDD 通过中断雌激素受体（ER）抑制排卵，和/或 孕激素受体（PR）介导的靶基因转录激活 卵巢卵泡中的基因。 一个具体的目标是确定细胞 通过定位芳烃在卵巢中TCDD作用的靶点 受体（Ah-Rs）和Ah-R核转位子（ARNTs），利用原位 未成熟大鼠卵巢在卵泡不同阶段的杂交 发育和排卵。 第二个具体目标是确定是否 TCDD在何种水平上改变ER和/或PR介导的信号通路 在卵巢里 卵巢中的ER和PR表达将通过 通过原位杂交或RT-PCR以及通过蛋白质分析进行mRNA分析 使用来自未成熟促性腺激素处理的大鼠卵巢的蛋白质印迹分析。 TCDD对雌、孕激素刺激靶点的抑制作用 通过检测Ers的结合能力来研究基因转录 和Prs到DNA（凝胶位移分析）和转录的能力， 目的基因的启动子。