ROLE OF ESTROGEN-REGULATED GENES DURING EARLY PREGNANCY

这项建议的长期目标是探索类固醇激素雌激素(E)如何在怀孕早期(植入前)调节子宫内膜的生长和分化过程，从而导致获得允许胚泡植入的接受状态。E通过调节特定靶基因的表达来发挥其细胞效应。然而，在生殖周期和妊娠的不同阶段，E调节基因的特性、表达谱和功能仍很不清楚。本研究的具体目的是：1.分离和鉴定在延迟着床过程中大鼠子宫中对强迫性E反应的调控基因。利用信使RNA差异显示方法分离和鉴定在大鼠子宫中表达受着床诱导剂量E诱导或抑制的mRNAs的cDNAs。为了确定新发现的E调节基因是否是潜在的着床调控因子，将通过Northern印迹、原位杂交和免疫组织化学方法分析其在早孕大鼠子宫中的时空表达。2.评估新发现的cDNA在着床中的功能作用。一项最新发展的反义技术将被用来调节完整大鼠子宫中的特定基因表达。这将涉及将针对每个候选基因的mRNA转录的反义寡核苷酸注射到植入前子宫中，以抑制该mRNA的稳定水平。如果这种干预也导致植入过程中的损害，它将允许在该基因和植入过程之间建立起功能联系。3.分析一个新的E调节基因ERG1在早期妊娠中的功能。最近，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)被分离出来。ERG1在卵巢周期和早期妊娠中以高度阶段特异性的方式在子宫表面上皮细胞中表达。为了确定ERG1在早期妊娠中的功能作用，我们将培育携带ERG1目标胚系突变的小鼠，并对其潜在的生殖表型进行分析，例如胚胎通过输卵管运输的障碍，上皮细胞形态和功能的缺陷，以及缺乏对植入的子宫接受性。这项拟议的研究将帮助我们确定在怀孕早期调节关键事件的分子。

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.