Morphogenesis of the paraxial mesoderm in mice

小鼠近轴中胚层的形态发生

• 批准号: 7143348
• 负责人: ANNA-KATERINA HADJANTONAKIS
• 金额: $ 39.31万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-07 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7143348
• 关键词: Wnt gene /protein; bioimaging /biomedical imaging; biological signal transduction; cell differentiation; cell migration; cell proliferation; congenital disorders; developmental genetics; embryogenesis; embryonic stem cell; gene mutation; genetically modified animals; green fluorescent proteins; histogenesis; laboratory mouse; mammalian embryology; mesoderm; molecular /cellular imaging; reporter genes

DESCRIPTION (provided by applicant): The broad aim of this project is to use a combination of molecular, embryological, genetic and optical techniques to understand the cellular and molecular events that direct the morphogenesis of mesoderm within the mouse embryo. Our central hypothesis is that Wnt3a is a key signaling molecule essential for several aspects of mesoderm specification and patterning. Recent advances in the isolation of proteins that naturally fluoresce, and the refinement of techniques for optical microscopy offer unprecedented opportunities for in vivo imaging to study cellular and molecular events directly within embryos. The application of optical imaging combined with a novel panel of transgenic reporter expressing mice that we have generated represent a unique platform for acquiring quantitative information on cell behavior and cell fate in vivo. We will exploit these reagents to investigate the dynamic cell behaviors integral to the morphogenesis of the paraxial mesoderm in wild type embryos and an allelic series of Wnt3a mutants that disrupt this process. The Specific Aims of this proposal are: (1) To use in vivo imaging to define the cell behaviors integral to the genesis of mesoderm within the primitive streak of the mouse embryo. (2) To use in vivo imaging to define the cell behaviors integral to the presomitic mesoderm. (3) To establish if paraxial mesoderm progenitor cells reside within the primitive streak of the mouse embryo. A role for signaling mediated by Wnt3a in these processes will be tested through use of the allelic series of Wnt3a mutants. This work will shed light on the cellular defects resulting from perturbations in mesoderm formation and will impact our understanding of the pathogenesis of human birth defects affecting the skeleton and musculature. Furthermore our work investigating mesodermal progenitor cells will contribute information towards the isolation of mesodermal stem cells, the determination of their intrinsic cell behaviors, their molecular signature and ultimately their directed differentiation for use in cell-based therapies.

描述（由申请人提供）：该项目的总体目标是结合分子、胚胎学、遗传和光学技术来了解指导小鼠胚胎内中胚层形态发生的细胞和分子事件。我们的中心假设是 Wnt3a 是对中胚层规范和模式的多个方面至关重要的关键信号分子。天然荧光蛋白质分离的最新进展以及光学显微镜技术的改进为体内成像直接研究胚胎内的细胞和分子事件提供了前所未有的机会。光学成像的应用与我们生成的新型转基因报告基因表达小鼠组相结合，代表了获取体内细胞行为和细胞命运定量信息的独特平台。我们将利用这些试剂来研究野生型胚胎中轴旁中胚层形态发生中不可或缺的动态细胞行为，以及破坏这一过程的一系列等位基因 Wnt3a 突变体。该提案的具体目标是：（1）利用体内成像来定义小鼠胚胎原条内中胚层发生所必需的细胞行为。 (2) 使用体内成像来定义前体中胚层的细胞行为。 (3) 确定近轴中胚层祖细胞是否存在于小鼠胚胎的原条内。将通过使用 Wnt3a 突变体的等位基因系列来测试 Wnt3a 介导的信号在这些过程中的作用。这项工作将揭示中胚层形成扰动引起的细胞缺陷，并将影响我们对影响骨骼和肌肉组织的人类出生缺陷发病机制的理解。此外，我们研究中胚层祖细胞的工作将为中胚层干细胞的分离、确定其内在细胞行为、分子特征以及最终用于细胞疗法的定向分化提供信息。