TROPHOBLAST CELL MOTILITY

滋养层细胞活力

• 批准号: 2612055
• 负责人: Ann E Sutherland
• 金额: $ 21.5万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-06 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2612055
• 关键词: cell adhesion; cell cycle proteins; cell differentiation; cell growth regulation; cell motility; embryo implantation; gene expression; genetic transcription; genetic transduction; guanine nucleotide binding protein; laboratory mouse; microinjections; phase contrast microscopy; polymerase chain reaction; posttranslational modifications; protein kinase; protein structure function; trophoblast; vertebrate embryology; video microscopy; video recording system

Implantation of the embryo into the uterus and the resultant formation of the placenta are unique and critical features of mammalian development. Implantation is mediated by the invasive behavior of the trophoblast cells, which differentiate from the trophectoderm cells of the blastocyst. Trophoblast invasive behavior is developmentally regulated; the precursor trophectoderm cells are quiescent epithelial cells which acquire motile invasive behavior with differentiation. The goals of this proposal are to examine and define the changes in cell behavior that accompany trophoblast differentiation, and, following the paradigms recently defined for the control of cell motility in cultured cells, to test the role of the Rho family of small GTP-binding proteins in the developmental regulation of trophoblast cell behavior. Time-lapse video analysis of embryos cultured in normal medium and in the presence of inhibitors will be used to determine when motile behavior is normally initiated, and at what point in development transcription and translation are required for its onset. The results will be compared to analyses of trophoblast adhesivity and molecular differentiation, to determine the relationship between these components of invasive behavior. These results will better define the process of trophoblast differentiation, and provide the foundation for interpreting and extending the transgenic experiments. Members of the Rho family of GTP-binding proteins have been shown in cultured cells to regulate formation of filopodial protrusions (Cdc42Hs), Iamellipodia (Rac) and stress fibers and focal contacts (Rho). To determine the role of these three proteins in the regulation of trophoblast cell motility, gene transfer through recombinant adenovirus infection will be used to overexpress dominant negative or constitutively active forms of each protein in preimplantation stage embryos. The dominant negative overexpression experiments will demonstrate whether each of the Rho family members is involved in the control of trophoblast motility, while the constitutively active overexpression experiments will address whether these proteins serve as control points for temporal regulation of trophoblast motility. The results of these experiments will clarify the mechanisms regulating trophoblast behavior and differentiation, and will provide a model both for later, less accessible developmental events, and for invasive transformation in other systems.

将胚胎植入子宫和结果形成 胎盘的独特和关键特征 发展。 植入是由侵入性行为介导的 滋养细胞，它与滋养外胚层细胞不同 胚泡。 滋养细胞的侵入性行为是发展的 受监管；前体的滋养剂细胞是静止的上皮 通过分化获取运动侵入性行为的细胞。 这 该建议的目标是检查和定义单元的变化 伴随滋养细胞分化的行为，并遵循 最近为控制培养的细胞运动而定义的范例 细胞，测试小型GTP结合蛋白的Rho家族的作用 在滋养细胞行为的发展调节中。 在正常培养基和中培养的胚胎的延时视频分析 抑制剂的存在将用于确定何时运动 行为通常是启动的，并且在发展的何时 发作需要转录和翻译。 结果 将与滋养细胞粘合剂和分子的分析进行比较 分化，以确定这些组件之间的关系 侵入性行为。这些结果将更好地定义 滋养细胞的分化，并为解释奠定了基础 并扩展转基因实验。 GTP结合蛋白的Rho家族的成员已显示 培养的细胞调节丝虫突起的形成 （cdc42hs），iamellipodia（RAC）和应力纤维和焦点接触 （Rho）。 确定这三种蛋白在调节中的作用 滋养细胞细胞运动，基因通过重组转移 腺病毒感染将用于过表达显性阴性或 在植入阶段的组成型活性形式 胚胎。 主要的负过表达实验将 证明每个Rho家族成员是否参与 控制滋养细胞运动，而组成型活动 过表达实验将解决这些蛋白质是否用作 滋养细胞运动的时间调节的控制点。 这 这些实验的结果将阐明调节的机制 滋养细胞的行为和差异化，并将提供一个模型 稍后，易于访问的发展事件和侵入性 其他系统中的转换。