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 最大化研究者研究奖续展申请的总体目标是推进 了解脊椎动物胚胎发生过程中的诱导和形态发生过程如何协调 确保由母体和合子基因产物控制的早期诱导过程。 建立胚胎极性和胚层，同时收敛和延伸 (C&E) 原肠胚形成运动 胚胎组织沿前后轴拉长，并向内侧变窄。 Wnt/平面细胞极性（Wnt/PCP）信号通路使间充质的形态和行为极化 我们之前提出 Wnt/PCP 信号传导充当细胞的角色。 相对于胚胎前后轴定向细胞的指南针，但指南针是如何调节的 在之前的资助期间，我们惊讶地发现有几种途径在起作用。 与 Wnt/PCP 平行以在 C&E 期间极化细胞我们还涉及 C&E 的另一个保守调节因子。 平面极性，Dachsous 非典型钙粘蛋白，对于许多过程至关重要，包括胚胎 通过促进微管动力学，我们投入了大量精力来生成裂解和轴规格。 新的遗传工具来探测 Wnt/PCP 指南针的机制，并启动了新的遗传筛选 原肠胚形成调节剂。 这些发现和工具激发了我们未来在三个研究主题上的研究。 有多少膜受体，包括我们在 Wnt/PCP 中涉及的 Gpr125 粘附 GPCR 指南针，在 C&E 过程中相互作用，在内源性 gpr125 基因座、蛋白质组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Wnt/PCP复合物的形成 在第二个主题中，我们将扩展对 Dachsous 的研究并调查它是否有效。 通过使用 Dachsous 内源标记来调节原肠胚形成运动。 借助 GFP，我们将进行蛋白质组学研究以分离内源性相互作用因子，并进行基因组编辑以定义 在一个平行的、无偏见的遗传方法中，我们将继续对它的关键区域进行有希望的遗传筛选。 母体和母体合子突变会损害胚胎发生和原肠胚形成。 Wnt/PCP 和 Dachsous 平面细胞极性调节剂的机制研究和公正的遗传筛选， 将推进 MIRA 任务并了解归纳和形态发生过程是如何进行的 由于这些途径的组成部分的突变导致了脊椎动物原肠胚形成过程中的协调。 流产、出生缺陷和疾病，我们的研究将提供对它们的理解和诊断的见解， 并促进疗法的开发。