TGFB EXPRESSION DURING EMBRYO RECEPTIVITY AND CELL DEATH

胚胎接受和细胞死亡过程中 TGFB 的表达

• 批准号: 3426731
• 负责人: JOY I MULHOLLAND
• 金额: $ 5万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1993-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3426731
• 关键词: cell death; cell type; cervical /vaginal smear; embryo implantation; endometrium; estradiol; estrus; female; gel electrophoresis; gene expression; gene induction /repression; genetic regulation; genetic transcription; hormone regulation /control mechanism; immunocytochemistry; laboratory rat; northern blottings; nuclear runoff assay; ovariectomy; progesterone; prostate; protein biosynthesis; transforming growth factors; uterus; western blottings

Although many events in the mammalian uterus are steroid hormone dependent, little is known about the cellular mechanisms through which estradiol and progesterone direct specific uterine functions. The proposed research will determine if certain actions of steroid hormones on different uterine cell types could be mediated by transforming growth factor beta (TGFB). Since it is not yet known if TGFB is regulated by steroid hormones in the uterus, synthesis of this protein will be examined during two important uterine functions which are directed by distinct hormonal milieux; receptivity to embryo attachment and cell death. TGFB transcription and synthesis will be examined in separated luminal epithelial and stromal cells from the rat uterus under hormonal conditions which give rise to each of these functions. Uterine receptivity will be induced in ovariectomized rats by a series of progesterone and estradiol injections which mimic the hormonal conditions of early pregnancy. Cell death will be induced by ovariectomy. TGFB transcription will be measured using northern and RNA slot-blot hybridization analyses of cytoplasmic RNA as well as transcription and nuclear run-off assays of newly transcribed RNA in cell nuclei. Synthesis of TGFB will be analyzed by gel-electrophoresis and immunoblotting of uterine cell proteins. The results of these experiments should demonstrate whether a change in TGFB synthesis is associated either with receptivity to embryo attachment or with cell death, if TGFB synthesis is regulated by steroid hormones in specific uterine cell types, and whether this regulation is transcriptional, posttranscriptional, or translational. If a role for TGFB in establishing receptivity for embryo implantation in the rat is indicated by this research, a firm precedent would be established for future investigations to determine if TGFB serves the same function in other mammals and if it is involved in human infertility.

尽管哺乳动物子宫中的许多事件都依赖于类固醇激素， 关于雌二醇和雌激素的细胞机制知之甚少。 黄体酮直接影响特定的子宫功能。 拟议的研究将 确定类固醇激素对不同子宫细胞是否有某些作用 类型可以通过转化生长因子β（TGFB）介导。 自从 目前尚不清楚 TGFB 是否受到子宫内类固醇激素的调节， 这种蛋白质的合成将在两个重要的子宫期间进行检查 由不同的荷尔蒙环境指导的功能；接受度 胚胎附着和细胞死亡。 TGFB的转录和合成将 在大鼠分离的管腔上皮细胞和基质细胞中进行检查 子宫处于荷尔蒙条件下，会产生这些 功能。 切除卵巢的大鼠可通过以下方法诱导子宫容受性 一系列黄体酮和 模拟早期荷尔蒙状况的雌二醇注射 怀孕。 卵巢切除术将诱导细胞死亡。 TGFB转录 将使用 Northern 和 RNA 狭缝印迹杂交分析进行测量 细胞质 RNA 以及转录和核径流测定 细胞核中新转录的RNA。 TGFB的合成将被分析 通过子宫细胞蛋白的凝胶电泳和免疫印迹。 这 这些实验的结果应该证明 TGFB 是否发生变化 合成与胚胎附着的接受性或 如果 TGFB 合成受类固醇激素调节，则细胞死亡 特定的子宫细胞类型，以及这种调节是否有效 转录、转录后或翻译。 如果 TGFB 发挥作用 在建立大鼠胚胎植入的接受性时表明 通过这项研究，将为未来建立一个坚实的先例 研究以确定 TGFB 是否在其他疾病中具有相同的功能 哺乳动物以及是否与人类不育有关。