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， 关于雌二醇和雌二醇通过细胞机制 孕酮指导特定的子宫功能。 拟议的研究将 确定类固醇激素对不同子宫细胞的某些作用 转化生长因子β（TGF-β）介导的。 以来 尚不清楚TGFB是否受子宫中类固醇激素的调节， 这种蛋白质的合成将在两个重要的子宫 由不同的激素环境指导的功能;对 胚胎附着和细胞死亡。 TGFB的转录和合成将 在来自大鼠的分离的管腔上皮细胞和基质细胞中进行检查 子宫激素条件下，引起这些每一个 功能协调发展的 在卵巢切除大鼠中，将通过以下方法诱导子宫容受性： 一系列的孕酮， 雌二醇注射，模拟早期的激素条件， 怀孕 卵巢切除术将诱导细胞死亡。 TGFB转录 将使用北方和RNA狭缝印迹杂交分析进行测量 细胞质RNA以及转录和核径流分析， 新转录的RNA。 对TGFB的合成进行了分析 通过凝胶电泳和子宫细胞蛋白质的免疫印迹。 的 这些实验的结果应该证明TGFB的变化是否 合成与胚胎附着的接受性有关， 与细胞死亡，如果TGFB的合成是由类固醇激素调节， 特定的子宫细胞类型，以及这种调节是否 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接受性方面， 通过这项研究，将为未来建立一个坚实的先例。 调查以确定TGFB是否在其他 哺乳动物以及是否与人类不育有关。

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.