ASPECTS OF UTERINE CELL CYCLE REGULATION IN IMPLANTATION

描述：（改编自申请人摘要）胚胎的过程 着床与子宫间质细胞蜕膜化有关， 这对怀孕的成功至关重要。蜕膜化过程是 其特征在于基质细胞生长并转化为蜕膜细胞， 以及由于核内复制而发生的基质细胞多倍性的形成 没有细胞分裂。细胞分裂的过程受到严格的控制， 细胞周期在两个特定的检查点，G1-S和G2-M。几种细胞 已知细胞周期蛋白（A、B、D和E型细胞周期蛋白）在以下方面起关键作用： 在这些过渡期间以特定的方式进行细胞生长。在这 关于这一点，PI最近已经证明，细胞周期蛋白D3的上调， 与子宫的蜕膜化密切相关。人们普遍认为， 细胞周期的动态控制通过细胞周期蛋白的相互作用来执行， 细胞周期蛋白依赖性激酶（cdk）和cdk抑制剂（CKI）。的目标 该修订的申请集中于细胞周期调节剂， 参与的发病和发展的蜕膜化方面的 植入过程。假设细胞周期调节因子是 在子宫中以时空方式表达， 互动在这些过程中发挥关键作用。具体目标是：（1） 表征细胞周期蛋白（A、B、D2和E）、cdks (cdk1 cdk 2、cdk 4和cdk 6）和CKIs（p27和p21）在着床期的表达 (days 1-8）野生型小鼠子宫; 2）研究细胞周期的作用 表皮生长因子EGF家族的作用 探讨了脑缺血再灌注损伤的机制和功能状态， 着床期间子宫中的cdks、相应的细胞周期蛋白和CKI， 在细胞周期蛋白D2、D3或p27缺失小鼠中的蜕膜化;和4）评估状态 细胞周期蛋白（A、B、D2、D3和E）、cdks（cdk 1、cdk 2、cdk 4和cdk 6）和CKIs（p27 和p21）在Hoxa-10缺失小鼠子宫中的表达。调查人员将描述 通过北方和原位杂交检测mRNA表达， 免疫组化、Western印迹和免疫沉淀。功能 细胞周期蛋白-cdk-CKI相关复合物的活性将通过体外 磷酸化活性。从这些研究中获得的结果将提供 关于关键细胞周期调节因子的潜在作用的重要见解 在着床和蜕膜化过程中的子宫生物学。由于这些 过程类似于伪恶性肿瘤，结果还应提供 与癌症生物学相关的基本信息。

DESCRIPTION: (Adapted from the applicant's abstract) The process of embryo implantation is associated with uterine stromal cell decidualization, an event that is critical to the success of pregnancy. The decidualization process is characterized by stromal cell growth and transformation into decidual cells, and formation of stromal cell polyploidy which occurs due to endoreduplication without cell division. The process of cellular division is tightly regulated in the cell cycle at two particular checkpoints, G1-S and G2-M. Several cell cyclins (A, B, D and E type cyclins) are known to play key roles with regard to cellular growth in a phase specific manner during these transitions. In this regard, the PI has recently demonstrated that the upregulation of cyclin D3 is tightly associated with decidualization of the uterus. It is well accepted that the dynamic control of the cell cycle is executed by an interplay of cyclins, cyclin-dependent kinases (cdks) and cdk inhibitors (CKIs). The objectives of this revised application are to focus on the cell cycle regulators that participate in the onset and progression of decidualization aspects of the implantation process. The hypothesis is that cell cycle regulators are expressed in the uterus in a spatiotemporal manner and their coordinated interactions play key roles in these processes. The specific aims are: 1) To characterize the spatiotemporal expression of cyclins (A, B, D2 and E), cdks (cdk1, cdk2, cdk4 and cdk6) and CKIs (p27 and p21) in the periimplantation (days 1-8) uterus of wild-type mice; 2) To study the role of cell cycle molecules in decidualization and the effects of the EGF family of growth factors in this process; 3) To study the regulation and functional status of cdks, respective cyclins and CKIs in the uterus during implantation and decidualization in cyclin D2, D3 or p27 null mice; and 4) To assess the status of cyclins (A, B, D2, D3 and E), cdks (cdk1, cdk2, cdk4 and cdk6) and CKIs (p27 and p21) in the uteri of Hoxa-10 null mice. The investigators will characterize mRNA expression by Northern and in situ hybridization and protein expression by immunohistochemistry, Western blotting and immunoprecipitation. The functional activity of cyclin-cdk-CKI associated complexes will be determined by in vitro phosphorylation activity. The results obtained from these studies will provide important insights regarding the potential roles of key cell cycle regulators in uterine biology during implantation and decidualization. Since these processes are analogous to pseudomalignancy, the results should also provide basic information relevant to cancer biology.