Interaction between blastocyst and Uterine endometrium

囊胚和子宫内膜之间的相互作用

• 批准号: 7802311
• 负责人: R. MICHAEL ROBERTS
• 金额: $ 28.74万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7802311
• 关键词: Animals; Binding; Biology; Cattle; Cell surface; Cells; Complementary DNA; Conceptus; Culture Media; Cytoplasmic Tail; Development; Embryo; Endometrial; Endometrium; Epithelial Cells; Epithelium; Estrous Cycle; Event; Evolution; Family; Florida; Funding; Genes; Goals; IFNAR2 gene; Implant; Inflammatory Response; Interferons; Knockout Mice; Length; Ligand Binding; Link; Mammalian Cell; Mammals; Missouri; Mothers; Mus; Mutation; Pathway interactions; Phenotype; Physiology; Placenta; Play; Pregnancy; Pregnancy loss; Procedures; Process; Proteins; Public Health; RNA Interference; Recruitment Activity; Research; Role; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Site; Staging; Stress; Testing; Time; Transcriptional Regulation; Universities; Uterus; Woman; Yeasts; biological adaptation to stress; blastocyst; corpus luteum; follow-up; homeodomain; implantation; insight; knock-down; pregnant; preimplantation; protein purification; receptor; response; secretory protein; stress activated protein kinase; tau interferon; transcription factor; trophoblast; yeast two hybrid system

DESCRIPTION (provided by applicant): Early pregnancy loss is a feature of all mammals and is particularly high in women. It is a major public health concern. More than 50% of normally conceived pregnancies fail in the period at about the time the embryo attaches to the uterus and begins to implant. Among the causes are developmental abnormalities of the embryo, weak or late signaling from the trophectoderm (the precursor of the placenta) and an ill- prepared maternal uterus. Here we shall study two aspects of conceptus-maternal relationship in early pregnancy. The first (goal 1) is to define the role of the transcription factor Cdx2 in trophectoderm specification and functional differentiation. Aim 1A is to confirm that Cdx2 plays a key role in the transcriptional control of the genes for the signaling molecule interferon-tau (IFN-T) and other genes characteristically up-regulated in trophectoderm. Aim 1B will test the hypothesis that Cdx2 is necessary in directing trophectoderm emergence in murine and bovine embryos and in maintaining a polarized epithelium. Aim 1C will confirm preliminary studies that Cdx2 is a marker of presumptive trophectoderm as early as the 2-cell stage of development. We shall also determine whether it is an analogous marker of cell fate in the bovine embryo. Goal 2 of this project is to define the function of Sin1, a newly discovered protein that associates with carboxyl end of the cytoplasmic domain of the IFN receptor subunit, IFNAR2. We hypothesize that maternal recognition of pregnancy involves a localized inflammatory response and the modulation of stress pathways in maternal endometrium at the site of implantation, and that Sin1 links the IFN response pathway to signal transduction pathways, including the stress-activated protein kinase (SAPK) pathway. Aim 2A is to define partners for Sin1 in endometrial signaling through the use of a yeast two-hybrid screen. Aim 2B will test the hypothesis that when IFN-T binds to the cell surface of uterine epithelial cells Sin 1 will be recruited, and this event will precede changes in the activities of stress-activated signal transduction pathways. Aim 2C will examine whether there is cellular relocation of Sin1 following IFN-T binding to the cell. Aim 2D will attempt to knock down Sin1 expression by RNAi and, by using this approach, determine whether Sin1 plays a role in some of the varied (pleiotropic) actions of type I IFN on mammalian cells. Aim 2E will explore whether the over-expression of different domains of the Sin1 protein can interfere with normal Sin1 function. Aim 2F is to create Sin1 null mice in order to gain a more complete insight into Sin1 function in the whole animal as well as in uterine endometrium. Together these studies are expected to provide information on two key events in early pregnancy, the formation of an early placenta and immediate maternal responses to the presence of a tiny embryo attaching to her uterine wall.

描述(申请人提供)：早孕丢失是所有哺乳动物的特征，在女性中尤为严重。这是一个重大的公共卫生问题。超过50%的正常妊娠在胚胎附着到子宫并开始植入的那段时间失败。其中的原因包括胚胎发育异常，滋养外胚层(胎盘的前身)发出的信号微弱或延迟，以及孕妇子宫准备不足。在这里，我们将研究怀孕早期母胎关系的两个方面。第一个(目标1)是确定转录因子CDX2在滋养外胚层规范和功能分化中的作用。目的1证实CDX2在滋养外胚层信号分子干扰素-tau(干扰素-T)和其他特征上调基因的转录调控中起关键作用。目的1B将验证这一假设，即在小鼠和牛胚胎中，CDX2在引导滋养外胚层的出现和维持极化上皮方面是必要的。目的1C将证实初步研究表明，早在2-细胞发育阶段，CDX2就可能是滋养外胚层的标志。我们还将确定它是否是牛胚胎中细胞命运的类似标记。该项目的目标2是确定SIN1的功能，SIN1是一种新发现的蛋白质，与干扰素受体亚单位IFNAR2的细胞质结构域的羧基末端相关。我们假设，母体对妊娠的识别涉及着床部位母体子宫内膜局部的炎症反应和应激通路的调节，SIN1将干扰素反应通路与信号转导通路联系起来，包括应激激活蛋白激酶(SAPK)通路。目标2A是通过使用酵母双杂交筛选来确定SIN1在子宫内膜信号传导中的伙伴。目的2B将验证这样一种假设，即当干扰素-T结合到子宫上皮细胞表面时，Sin1将被招募，这一事件将先于应激激活信号转导通路的活性变化。Aim 2C将检测在干扰素-T与细胞结合后是否存在SIN1的细胞重新定位。目的2D试图通过RNAi抑制SIN1的表达，并通过这种方法来确定SIN1是否在I型干扰素对哺乳动物细胞的一些不同的(多效性)作用中发挥作用。目的2E探讨SIN1蛋白不同结构域的过度表达是否会干扰SIN1的正常功能。目的2F建立SIN1缺失小鼠，以便更全面地了解SIN1在整个动物体内以及子宫内膜中的功能。这些研究预计将提供关于早期怀孕的两个关键事件的信息，即早期胎盘的形成和母亲对附着在子宫壁上的微小胚胎的即时反应。