Body Plan Formation in Early Mouse Embryo

早期小鼠胚胎的身体计划形成

• 批准号: 6744094
• 负责人: YUSUKE MARIKAWA
• 金额: $ 18.4万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-08 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6744094
• 关键词: DNA binding protein; biological models; biological signal transduction; biomarker; cadherins; cell adhesion; cell differentiation; cell growth regulation; centrifugation; cytoplasm; cytoskeleton; early embryonic stage; egg /ovum; embryogenesis; embryonic stem cell; gene expression; laboratory mouse; leptin; membrane transport proteins; molecular dynamics; protein localization; protein structure function; trophoblast

DESCRIPTION: (Scanned from the applicant's abstract) The basic architecture of animal bodies is established during early embryogenesis. Compared to other vertebrate systems, little is known about how a mammalian egg creates the basic body plan. The major goal of this study is to elucidate the molecular mechanisms responsible for cell fate determination and body axis formation in the mouse embryo. The first specific aim of this study is to identify cytoplasmic and membrane components in the egg that are involved in the formation of the inner cell mass (ICM) and the trophectoderm (TE). Formation of the ICM and TE is the first cell differentiation event in mouse development. We will use lineage-specific molecular markers to identify the differentiation of these two distinct cell types in embryos treated with various inhibitors of the cytoskeleton, membrane trafficking and cell adhesion. We will also investigate the role of egg cytoplasmic components in the ICM and TE development. For this study, we will exploit the centrifugation techniques, which have been successfully used to identify developmentally important factors in the egg of other vertebrates and chordates. The second aim is to analyze the function of localized molecules in the egg, namely Stat3 and Leptin proteins. Since both Stat3 and Leptin act as regulators of cell differentiation and growth in many systems, we hypothesize that the localized distribution of Stat3 and Leptin in the egg plays an important role in the establishment of the body plan. We will test this hypothesis by overexpressing Stat3 and Leptin in the mouse egg to disturb their normal distribution. We will also use constitutive active and dominant negative constructs of Stat3 to examine the function of the localized Stat3 in more detail. The third aim is to define the role of the Wnt/b-catenin signaling pathway in body axis formation in the embryo. Although the signaling pathway is suggested to be involved in axis formation in the mouse embryo, its mode of action is not as clear as in other vertebrate systems, specifically the frog. We will define the role of the Wnt/b-catenin signaling pathway in the mouse axis formation in more detail by introducing inhibitory constructs of the signaling pathway, and by locally activating the signaling pathway in early embryos. The studies proposed in this application should enable us to better understand the molecular mechanisms of the earliest critical events in mammalian body plan formation, and provide clues to resolve many problems associated with human birth defects as well as biomedical technology.

描述：（从申请人的摘要扫描）的基本架构 动物的身体是在早期胚胎发育过程中形成的。相比其他 在脊椎动物系统中，很少有人知道哺乳动物的卵是如何产生基本的 身体计划。本研究的主要目的是阐明 细胞命运决定和体轴形成的机制 老鼠胚胎本研究的第一个具体目标是确定 卵细胞中的细胞质和膜成分参与了 内细胞团（ICM）和滋养外胚层（TE）的形成。形成 ICM和TE是小鼠发育中的第一个细胞分化事件。我们 将使用谱系特异性分子标记来鉴定 这两种不同的细胞类型在胚胎处理与各种抑制剂的 细胞骨架、膜运输和细胞粘附。我们亦会研究 卵细胞质成分在ICM和TE发育中的作用。为此 研究，我们将利用离心技术，这已经是 成功地用于确定发育的重要因素，在鸡蛋， 其他脊椎动物和脊索动物。第二个目的是分析 在鸡蛋中的局部分子，即Stat 3和瘦素蛋白。由于两 Stat 3和Leptin在许多肿瘤细胞中作为细胞分化和生长的调节因子发挥作用。 系统中，我们假设Stat 3和Leptin的局部分布， 鸡蛋在身体计划的制定中起着重要的作用。我们将 通过在小鼠卵细胞中过表达Stat 3和Leptin来验证这一假设， 扰乱其正常分布。我们还将使用结构活性和 Stat 3的显性负性结构，以检查定位的 第3章更详细第三个目的是确定Wnt/b-连环蛋白的作用， 胚胎中轴形成的信号通路。虽然信号 通路被认为参与了小鼠胚胎中轴的形成， 作用模式不像其他脊椎动物系统那样清楚，特别是 青蛙我们将明确Wnt/β-连环蛋白信号通路在细胞凋亡中的作用。 通过引入抑制性构建体，更详细地描述了小鼠轴形成。 信号通路，并通过早期局部激活信号通路 胚胎本申请中提出的研究应使我们能够更好地 了解最早的关键事件的分子机制， 哺乳动物的身体计划的形成，并提供线索，解决许多问题 与人类出生缺陷以及生物医学技术有关。