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: 3426730
• 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/3426730
• 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是类固醇激素依赖的，但 关于雌二醇和雌二醇和的细胞机制知之甚少 孕酮直接特异性子宫功能。 拟议的研究将 确定类固醇激素是否在不同子宫细胞上的某些作用 可以通过转化生长因子β（TGFB）来介导类型。 自从 尚不清楚TGFB是否受子宫中类固醇激素调节， 该蛋白的合成将在两个重要的子宫中检查 由不同的荷尔蒙环境导演的功能；接受 胚胎附着和细胞死亡。 TGFB转录和合成将 在大鼠的分离的腔上皮和基质细胞中检查 子宫在荷尔蒙条件下产生每一种 功能。 子宫接受能力将在卵巢切除大鼠中诱导 一系列孕酮和 雌二醇注射，模仿早期的荷尔蒙条件 怀孕。 细胞死亡将由卵巢切除术诱导。 TGFB转录 将使用北部和RNA插槽印刷杂交分析来测量 细胞质RNA以及转录和核径流测定法 细胞核中新转录的RNA。 将分析TGFB的合成 通过凝胶 - 电泳和子宫细胞蛋白的免疫印迹。 这 这些实验的结果应证明TGFB是否发生变化 合成与胚胎附着的接受性或 在细胞死亡中，如果TGFB合成受到类固醇激素的调节 特定的子宫细胞类型以及该法规是否为 转录，转录后或翻译。 如果TGFB的角色 在建立对大鼠胚胎植入的接受度时 通过这项研究，将建立企业先例 调查以确定TGFB在其他方面是否具有相同的功能 哺乳动物以及它与人类不孕症有关。