Interaction between blastocyst and Uterine endometrium

囊胚和子宫内膜之间的相互作用

• 批准号: 7416588
• 负责人: R. MICHAEL ROBERTS
• 金额: $ 29.03万
• 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 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7416588
• 关键词: Animals; Binding; Biology; Bos taurus; 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; Localized; Mammalian Cell; Mammals; Missouri; Mothers; Mus; Mutation; Pathway interactions; Patient currently pregnant; 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; Standards of Weights and Measures; Stress; Testing; Time; Transcriptional Regulation; Universities; Uterus; Woman; Yeasts; biological adaptation to stress; blastocyst; corpus luteum; follow-up; homeodomain; implantation; insight; knock-down; 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（由申请人提供）：早孕丢失是所有哺乳动物的特征，在女性中尤其高。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时期失败。原因包括胚胎发育异常、滋养外胚层（胎盘的前身）信号微弱或延迟以及母体子宫准备不足。在这里，我们将研究两个方面的概念，母体关系在怀孕早期。第一个（目标1）是定义的作用，转录因子Cdx 2在滋养外胚层的规范和功能分化。目的1A是证实Cdx 2在信号分子干扰素-tau（IFN-T）的基因和滋养外胚层中特征性上调的其他基因的转录控制中起关键作用。目的1B将测试的假设，Cdx 2是必要的，在指导滋养外胚层出现在小鼠和牛胚胎和维持极化上皮。目的1C将证实Cdx 2是早在发育的2细胞阶段的假定滋养外胚层的标志物的初步研究。我们还将确定它是否是牛胚胎中细胞命运的类似标记。本项目的目标2是确定Sin 1的功能，Sin 1是一种新发现的蛋白质，与IFN受体亚单位IFNAR 2的胞质结构域的羧基端相关。我们推测，母亲识别怀孕涉及一个本地化的炎症反应和调制的压力途径在母体子宫内膜在植入的网站，和Sin 1连接的干扰素反应途径的信号转导途径，包括应激活化蛋白激酶（SAPK）途径。目的2A是通过使用酵母双杂交筛选来确定子宫内膜信号转导中Sin 1的伴侣。目的2B将验证以下假设：当IFN-T结合到子宫上皮细胞的细胞表面时，Sin 1将被募集，并且该事件将先于应激激活的信号转导通路的活性的变化。目的2C将检查在IFN-T结合到细胞后是否存在Sin 1的细胞重定位。目的2D将试图敲低Sin 1表达的RNAi，并通过使用这种方法，确定是否Sin 1发挥作用，在一些不同的（多效性）行动的I型干扰素对哺乳动物细胞。目的2 E探讨Sin 1蛋白不同结构域的过表达是否会干扰Sin 1的正常功能。目的2F是建立Sin 1基因敲除小鼠，以便更全面地了解Sin 1在整个动物以及子宫内膜中的功能。这些研究预计将共同提供关于妊娠早期两个关键事件的信息，即早期胎盘的形成和母体对附着在子宫壁上的微小胚胎的立即反应。