Inductive and morphogenetic processes shaping the zebrafish embryonic axes

塑造斑马鱼胚胎轴的诱导和形态发生过程

• 批准号: 10207136
• 负责人: LILIANNA SOLNICAKREZEL
• 金额: $ 67.32万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10207136
• 关键词: Adhesions; Award; Behavior; Cadherins; Cell Shape; Cells; Complex; Congenital Abnormality; Development; Diagnosis; Disease; Embryo; Embryonic Development; Ensure; Funding; Future; G-Protein-Coupled Receptors; Gastrula; Genetic; Genetic Screening; Germ Layers; Goals; Impairment; Membrane; Mesenchymal; Microtubules; Mission; Molecular; Morphogenesis; Morphology; Movement; Mutation; Pathway interactions; Pattern; Process; Proteomics; Research; Research Personnel; Shapes; Signal Pathway; Signal Transduction; Spontaneous abortion; Testing; Time; Work; Zebrafish; embryo tissue; embryonic cleavage; experimental study; gastrulation; gene product; genetic approach; genome editing; insight; planar cell polarity; polarized cell; receptor; therapy development; tool

ABSTRACT The overall goal of this Maximizing Investigator’s Research Award renewal application is to advance understanding of how inductive and morphogenetic processes during vertebrate embryogenesis are coordinated to ensure normal development. Early inductive processes controlled by maternal and zygotic gene products establish embryonic polarity and germ layers, while convergence and extension (C&E) gastrulation movements elongate embryonic tissues down the anteroposterior axis and narrow them mediolaterally. The noncanonical Wnt/Planar Cell Polarity (Wnt/PCP) signaling pathway polarizes morphologies and behaviors of mesenchymal gastrula cells that shape embryonic body. We previously proposed that Wnt/PCP signaling acts as a cellular compass that orients cells with respect to the anteroposterior embryonic axis, but how the compass is regulated is not understood. During the previous funding period, we surprisingly found that several pathways work in parallel to Wnt/PCP to polarize cells during C&E. We also implicated in C&E another conserved regulator of planar polarity, Dachsous atypical cadherin, which is essential for many processes, including embryonic cleavages and axis specification, by promoting microtubule dynamics. We invested significant effort in generating new genetic tools to probe the mechanisms of the Wnt/PCP compass and initiated a genetic screen for new gastrulation regulators. These findings and tools motivate our future studies in three research themes. In the first, we will investigate how numerous membrane receptors, including Gpr125 adhesion GPCR, which we implicated in the Wnt/PCP compass, interact during C&E. Using precise mutations and tags in the endogenous gpr125 locus, proteomic and genetic experiments, we will test whether Gpr125 promotes formation of Wnt/PCP complexes composed of select components. In the second theme, we will extend our studies of Dachsous and investigate whether it regulates gastrulation movements by promoting microtubule dynamics. Using Dachsous endogenously tagged with GFP, we will carry out proteomic studies to isolate endogenous interactors, and genome editing to define its critical regions. In a parallel unbiased genetic approach, we will continue a promising genetic screen for maternal and maternal-zygotic mutations that impair embryogenesis and gastrulation. Altogether, our mechanistic studies of Wnt/PCP and Dachsous regulators of planar cell polarity and unbiased genetic screens, will both advance the MIRA mission and understanding how inductive and morphogenetic processes are coordinated during vertebrate gastrulation. As mutations in the components of these pathways cause miscarriages, birth defects and diseases, our studies will provide insights into their understanding and diagnosis, and facilitate development of therapies.

抽象的 这项最大化调查员研究奖的续签申请的总体目标是促进 了解脊椎动物胚胎发生过程中的感应和形态发生过程如何协调 确保正常发展。由母体和二元基因产物控制的早期归纳过程 胚胎极性和细菌层，而收敛和延伸（C＆E）过度运动 延伸到前后轴的胚胎组织，并在中等范围内窄。非规范 Wnt/Planar细胞极性（WNT/PCP）信号通路途径极化间质的形态和行为 形成胚胎体的胃细胞。我们先前提出WNT/PCP信号作为细胞 指示细胞相对于前后胚胎轴的指南针，但如何调节指南针 不了解。在上一个资金期间，我们出人意料地发现，有几条途径在 与Wnt/PCP平行在C＆E期间偏振细胞。我们还在C＆E中实施了另一个组成的监管机构 平面极性，硫硫酸非典型钙粘蛋白，这对于许多过程至关重要，包括胚胎 通过促进微管动力学来切割和轴规范。我们投入了巨大的努力来产生 新的遗传工具来探测Wnt/PCP指南针的机制，并启动了新的遗传屏幕 过度调节器。 这些发现和工具激励我们以三个研究主题的未来研究。首先，我们将调查 我们在WNT/PCP中实现的膜受体有多少，包括GPR125粘合剂GPCR 指南针，在C＆E期间互动。在内源性GPR125基因座，蛋白质组学中使用精确的突变和标签 和基因实验，我们将测试GPR125是否促进了由Wnt/PCP组成的形成 选择组件。在第二个主题中，我们将扩展对采率的研究，并调查它是否 通过促进微管动力学来调节过失运动。使用二豆式内源标记 使用GFP，我们将进行蛋白质组学研究以分离内源性相互作用，并进行基因组编辑以定义 它的关键地区。在平行的公正遗传方法中，我们将继续一个有希望的遗传筛选 损害胚胎发生和过失的母体和母体 - 杂种突变。总共，我们的 Wnt/PCP和平面细胞极性和无偏基遗传筛查的辅助剂的机械研究， 既可以促进MIRA任务，又要理解归纳和形态发生过程 在脊椎动物过口期间协调。作为这些途径组件中的突变引起的 流产，出生缺陷和疾病，我们的研究将提供有关其理解和诊断的见解， 并促进疗法的发展。