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.

将胚胎植入子宫并形成 胎盘的结构是哺乳动物独特且重要的特征 发展。 植入是由侵入行为介导的 滋养层细胞，由滋养外胚层细胞分化而来 囊胚。 滋养层的侵袭行为是发育性的 受监管；前体滋养外胚层细胞是静止的上皮细胞 通过分化获得运动侵袭行为的细胞。 这 该提案的目标是检查和定义细胞中的变化 伴随滋养层分化的行为，并且遵循 最近定义的用于控制培养细胞运动的范式 细胞，测试小 GTP 结合蛋白 Rho 家族的作用 滋养层细胞行为的发育调节。 对正常培养基和培养皿中培养的胚胎进行延时视频分析 抑制剂的存在将用于确定何时运动 行为通常是在什么时候开始的？ 它的发生需要转录和翻译。 结果 将与滋养层粘附性和分子分析进行比较 微分，以确定这些组件之间的关系 的侵略行为。这些结果将更好地定义过程 滋养层分化，并为解释提供基础 并扩大转基因实验。 GTP 结合蛋白 Rho 家族的成员已在 培养细胞调节丝状伪足突起的形成 (Cdc42Hs)、板状伪足 (Rac) 和应力纤维和焦点接触 （罗）。 确定这三种蛋白质在调节中的作用 滋养层细胞运动、通过重组进行基因转移 腺病毒感染将用于过度表达显性失活或 植入前阶段每种蛋白质的组成型活性形式 胚胎。 显性阴性过度表达实验将 证明 Rho 家族的每个成员是否都参与了 控制滋养层运动，而组成性活跃 过表达实验将解决这些蛋白质是否充当 滋养层运动的时间调节的控制点。 这 这些实验的结果将阐明调节机制 滋养层行为和分化，并将提供一个模型 对于后来的、不易接近的发育事件，以及侵入性的 其他系统的改造。