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: 9918423
• 负责人: LILIANNA SOLNICAKREZEL
• 金额: $ 63.29万
• 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 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9918423
• 关键词: Adhesions; Anterior; Behavior; Cadherins; Calcium Signaling; Cell Polarity; Cell membrane; Cells; Complex; Congenital Abnormality; Cytoskeleton; Development; Diagnosis; Disease; Dorsal; Embryo; Embryonic Development; Ensure; Fatty acid glycerol esters; G-Protein-Coupled Receptors; Genes; Germ Layers; Individual; Link; Membrane; Mesenchymal; Microtubules; Morphogenesis; Movement; Mutation; Pathway interactions; Pattern; Phenotype; Phosphotransferases; Process; Proteins; Research; Role; Shapes; Signal Transduction; Spontaneous abortion; System; Testing; Time; Vertebrates; Work; Zebrafish; beta catenin; chemokine; driving behavior; gastrulation; interest; loss of function; mutant; planar cell polarity; polarized cell; public health relevance; therapy development; zygote

 DESCRIPTION (provided by applicant): How does a vertebrate animal body develop from a single cell is an overarching question of my research. Despite much progress, many basic mechanisms underlying embryogenesis remain poorly understood. I am particularly interested how the inductive and morphogenetic processes that establish the embryonic body plan are coordinated in space and time to ensure normal development. Specification of anteroposterior and dorsoventral embryonic axis in the zebrafish zygote involves microtubule dependent transport of factors promoting dorsal Wnt/β-catenin activity, which patterns germ layers and coordinates gastrulation movements. We showed that Ccl19.1 chemokine signaling through Ccr7 G protein-coupled receptor (GPCR) limits axis formation by downregulating β-catenin. Using mutations in ccl19.1 and ccr7 zebrafish genes we will delineate the pathways through which they regulate calcium signaling and limit axis formation. We recently discovered that Dachsous (Dchs) cadherins regulate cytoskeleton in the zygote and during axis formation. Whereas Dchs is commonly known to interact with Fat cadherin and Fjx1 kinase, we will characterize Fat/Fjx1-dependent and independent roles of Dchs. We will identify proteins interacting with Dchs to define its functional links to cytoskeleton and other effectors. During gastrulation, convergence and extension (C&E) movements elongate the germ layers anteroposteriorly and narrow them dorsoventrally. Wnt/Planar Cell Polarity (Wnt/PCP) signaling is a conserved regulator of polarized cell behaviors driving C&E in vertebrates. Our studies posit that Wnt/PCP signaling works during gastrulation as a cellular compass coordinating polarity and behaviors of individual cells with anteroposterior embryonic polarity, in part by asymmetric distribution of PCP components at anterior and posterior cell membranes. To understand how the planar asymmetries of PCP components arise in dynamic mesenchymal cells, we will test the hypothesis that adhesion GPCR, Gpr125, a new PCP component, promotes formation of the posteriorly enriched membrane PCP complexes. We will determine whether Dchs works with Fat and Fjx1 during gastrulation to polarize microtubules and regulate Wnt/PCP signaling-dependent cell polarity. Finally, we will test whether the Dchs/Fat/Fjx1 system provides a mechanism to link Wnt/PCP signaling to global embryo patterning. To uncover additional chemokine GPCRs involved in axis formation and gastrulation, we will interrogate functions of the subset of 21 chemokine GPCRs expressed during embryogenesis by characterizing loss-of-function single and compound mutant phenotypes. As mutations in the components of the Wnt, Dchs/Fat and GPCR pathways cause miscarriages, birth defects and diseases; our studies can advance their understanding, diagnosis and facilitate development of therapies.

 描述（由申请人提供）：脊椎动物身体如何从单细胞发育是我研究的一个首要问题，尽管取得了很大进展，但胚胎发生的许多基本机制仍然知之甚少，我特别感兴趣的是诱导和形态发生过程。建立在空间和时间上协调的胚胎身体计划，以确保斑马鱼受精卵的前后和背腹胚胎轴的正常发育涉及微管依赖的因子运输。我们发现，Ccl19.1 趋化因子通过 Ccr7 G 蛋白偶联受体 (GPCR) 信号通过下调 β-catenin 来限制轴的形成。 1 和 ccr7 斑马鱼基因，我们将描述它们调节钙信号传导和限制轴形成的途径。我们最近发现了 Dachsous (Dchs) 钙粘蛋白。众所周知，Dchs 与脂肪钙粘蛋白和 Fjx1 激酶相互作用，在受精卵中调节细胞骨架，我们将鉴定与 Dchs 相互作用的蛋白质，以确定其功能联系。在原肠胚形成期间，收敛和伸展（C&E）运动使胚层前后伸长并变窄。我们的研究表明，Wnt/平面细胞极性 (Wnt/PCP) 信号是驱动脊椎动物 C&E 的极化细胞行为的保守调节因子，在原肠胚形成期间作为细胞指南针协调单个细胞的极性和行为。胚胎极性，部分通过 PCP 成分在前细胞膜和后细胞膜的不对称分布来了解 PCP 成分的平面不对称性。动态间充质细胞中产生的，我们将测试粘附 GPCR、Gpr125（一种新的 PCP 成分）促进后部富集膜 PCP 复合物的形成的假设。我们将确定 Dchs 在原肠胚形成过程中是否与 Fat 和 Fjx1 一起作用以极化微管并调节 Wnt。 /PCP信号依赖的细胞极性最后，我们将测试Dchs/Fat/Fjx1系统是否提供了链接Wnt/PCP信号的机制。为了揭示参与轴形成和原肠胚形成的其他趋化因子 GPCR，我们将通过表征功能缺失的单一和复合突变体表型来探究胚胎发生过程中表达的 21 种趋化因子 GPCR 的功能。 Wnt、Dchs/Fat 和 GPCR 通路导致流产、出生缺陷和疾病；我们的研究可以增进对它们的理解、诊断并促进治疗方法的开发。