ROLE OF ESTROGEN-REGULATED GENES DURING EARLY PREGNANCY

本研究的长期目标是探索类固醇激素雌激素(E)如何调节妊娠早期（着床前）子宫内膜的生长和分化过程，从而获得允许囊胚着床的接受状态。E通过调节特定靶基因的表达来发挥其细胞作用。然而，在生殖周期和妊娠的不同阶段，e调节基因的身份、表达谱和功能在很大程度上仍然未知。本建议的具体目的是：1。分离并鉴定延迟着床期间大鼠子宫内对中性E的调控基因。我们将采用信使RNA差异显示法分离和鉴定在诱导e着床剂量下大鼠子宫中诱导或抑制mrna表达的cdna。为了确定新鉴定的e调控基因是否为着床的潜在调节因子，我们将采用Northern blotting、原位杂交和免疫组织化学方法分析其在妊娠早期大鼠子宫中的时空表达。2. 评估新鉴定的cdna在植入中的功能作用。一项新开发的反义技术将用于调节完整大鼠子宫内特定基因的表达。这将涉及将针对每个候选基因mRNA转录物的反义寡脱氧核苷酸施用到着床前子宫，以抑制该mRNA的稳态水平。如果这种干预也导致植入损伤，它将允许在该基因和植入过程之间建立功能联系。3. 分析一种新的e调控基因ERG1在妊娠早期的功能。在子宫和输卵管这两个关键的生殖组织中，一个受E严格调控的新基因（ERG1）最近被分离出来。在卵巢周期和妊娠早期，ERG1在子宫表面上皮中以高度分期特异性的方式表达。为了确定ERG1在妊娠早期的功能作用，研究人员将培养具有ERG1靶向种系突变的小鼠，并分析其潜在的生殖表型，如胚胎通过输卵管运输障碍、上皮细胞形态和功能缺陷以及子宫对着床缺乏接受性。这项研究将帮助我们确定介导妊娠早期关键事件的E调节的分子。

The long-term goal of this proposal is to explore how the steroid hormone estrogen (E) regulates growth and differentiation processes in the endometrium during early pregnancy (preimplantation period), which lead to acquisition of the receptive state that allows blastocyst implantation. E exerts its cellular effects by regulating the expression of specific target genes. The identity, profile of expression, and function of the E-regulated genes at various stages of the reproductive cycle and pregnancy, however, remain largely unknown. The specific aims of this proposal are to: 1. Isolate and identify genes that are regulated in response to nidatory E in rat uterus during delayed implantation. The messenger RNA differential display method will be used to isolate and identify the cDNAs representing mRNAs whose expression is induced or repressed in rat uterus in response to an implantation-inducing dose of E. To determine whether the newly identified E-regulated genes are potential modulators of implantation, their spatio-temporal expression in rat uterus during early pregnancy will be analyzed by Northern blotting, in situ hybridization, and immunohistochemistry. 2. Assess the functional roles of the newly identified cDNAs in implantation. A recently developed antisense technology will be used to regulate specific gene expression in intact rat uterus. This will involve administration of antisense oligodeoxynucleotides targeted against mRNA transcript of each candidate gene into the preimplantation uterus to suppress the steady state level of that mRNA . If this intervention also leads to an impairment in implantation, it will allow the establishment of a functional link between this gene and the implantation process. 3. Analyze the function of a novel E-regulated gene, ERG1, in early pregnancy. A novel gene (ERG1) that is tightly regulated by E in two key reproductive tissues, the uterus and oviduct, has recently been isolated. ERG1 is expressed in the surface epithelium of the uterus in a highly stage-specific manner during the ovarian cycle and early pregnancy. To determine the functional role of ERG1 during early pregnancy, mice harboring a targeted germ line mutation of ERG1 will be developed and analyzed for potential reproductive phenotypes, such as impairment in embryo transport through the oviduct, defects in epithelial cell morphology and function, and lack of uterine receptivity for implantation. The proposed research will help us to identify molecules that mediate E regulation of critical events during early pregnancy.