ROLE OF ESTROGEN-REGULATED GENES DURING EARLY PREGNANCY

该提案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（植入前期）子宫内膜的生长和分化过程，从而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发挥其细胞效应。 然而，在生殖周期和妊娠的各个阶段，E调节基因的身份、表达谱和功能在很大程度上仍然未知。 这项建议的具体目标是：1.分离并鉴定在延迟着床过程中大鼠子宫中响应于nidorative E的调控基因。 信使RNA差异显示方法将用于分离和鉴定代表mRNA的cDNA，这些mRNA的表达在大鼠子宫中响应于诱导剂量的E. 为了确定新鉴定的E调节基因是否是潜在的着床调节因子，将通过北方印迹、原位杂交和免疫组织化学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在着床中的功能作用。 最近发展的反义技术将用于调控完整大鼠子宫中特定基因的表达。这将涉及将靶向每个候选基因的mRNA转录物的反义寡脱氧核苷酸施用到植入前子宫中以抑制该mRNA的稳态水平。如果这种干预也导致植入障碍，它将允许在该基因和植入过程之间建立功能联系。 3.分析一个新的E调节基因ERG 1在早期妊娠中的功能。 一个新的基因（ERG 1），在两个关键的生殖组织，子宫和输卵管，是紧密调控的E，最近被分离出来。 ERG1在卵巢周期和妊娠早期以高度阶段特异性的方式在子宫表面上皮中表达。 为了确定ERG 1在早期妊娠期间的功能作用，将开发携带ERG 1的靶向生殖系突变的小鼠，并分析其潜在的生殖表型，例如通过输卵管的胚胎运输受损、上皮细胞形态和功能缺陷以及缺乏植入的子宫容受性。 拟议的研究将帮助我们确定在怀孕早期介导E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.