TGFB EXPRESSION DURING EMBRYO RECEPTIVITY AND CELL DEATH

虽然哺乳动物子宫中的许多事件是类固醇激素依赖的， 目前对雌二醇和雌二醇单抗的细胞机制知之甚少。 黄体酮直接特异地发挥子宫功能。拟议的研究将 确定类固醇激素对不同子宫细胞的某些作用 类型可通过转化生长因子β(TGFb)来调节。自.以来 目前尚不清楚TGFb是否受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 由不同的荷尔蒙环境指导的功能；对 胚胎附着和细胞死亡。TGFb的转录和合成将 在分离的大鼠子宫腔上皮细胞和基质细胞中进行检测 子宫处于荷尔蒙状态下，会导致这些 功能。去卵巢大鼠子宫容受性的实验研究 一系列黄体酮和 雌二醇注射模拟早期的荷尔蒙状况 怀孕了。卵巢切除会导致细胞死亡。TGFb转录 将使用Northern和RNA插槽印迹杂交分析进行测量 细胞质RNA以及转录和核流失分析 在细胞核中新转录的RNA。TGFb的合成将被分析 通过凝胶电泳法和免疫印迹法检测子宫细胞蛋白。这个 这些实验的结果应该证明，TGFb的变化 合成与对胚胎附着的接受性或 在细胞死亡的情况下，如果TGFb的合成受到类固醇激素的调节 特定的子宫细胞类型，以及这一规定是否 转录的、转录后的或翻译的。如果TGFb的角色 在建立大鼠胚胎植入的感受性方面， 通过本文的研究，将为今后的研究奠定坚实的先例。 调查以确定TGFb是否在其他 如果它与人类不孕不育有关。

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.