Toddler and Apelin Signaling During Zebrafish Gastrulation

斑马鱼原肠胚形成过程中的幼儿和 Apelin 信号传导

• 批准号: 8784314
• 负责人: Megan L Norris
• 金额: $ 3.49万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8784314
• 关键词: Address; Adhesions; Affect; Behavior; Cardiovascular system; Cell Adhesion; Cell Adhesion Molecules; Cells; Complex; Cues; Defect; Development; Disease; Drug Targeting; E-Cadherin; Embryo; Embryo Deaths; Embryonic Development; Erinaceidae; Failure; Fibroblast Growth Factor; G-Protein-Coupled Receptors; Gene Expression; Generations; Genes; Genetic; Genetic Engineering; Genetic Epistasis; Germ Layers; Heart; Human; Immigration; Immunohistochemistry; Knowledge; Label; Laboratories; Ligands; Light; Messenger RNA; Microscopy; Molecular; Morphology; Movement; Names; Neurologic; Organ failure; Organism; Pathway interactions; Peptides; Phenotype; Process; Regulation; Reporter; Reporter Genes; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Syndrome; System; Testing; Therapeutic; Time; Toddler; Transplantation; Vertebrates; Western Blotting; Work; Zebrafish; cell motility; cell type; dosage; gastrulation; migration; mutant; novel; overexpression; public health relevance; receptor; research study; spatiotemporal

DESCRIPTION (provided by applicant): Vertebrate development depends on precise proliferation, migration and specification of cells to generate distinct and complex parts of an organism. Failure in any of these processes can cause defects such as organ failure, neurological syndromes or death of the embryo. Despite the importance of these processes, their pathways and regulators are not completely understood. This project will address this disparity by characterizing Toddler, a novel, highly conserved secreted peptide involved in embryogenesis and required for proper migration of the endodermal and mesodermal germ layers during gastrulation in zebrafish. Previous work in the laboratory demonstrated that Toddler regulates cell migration, though the change in cellular behavior and molecular mechanism stimulated by Toddler is not known. This work will test the hypothesis that Toddler is required for cell internalization during gastrulation (Aim 1) due to its regulation of E-cadherin (Aim 2) and partially overlaps in function with the known secreted peptide Apelin (Aim 3). First, genetic engineering and microscopy analysis will be used to label and track Toddler-responsive cells during development to identify the specific cellular behavior affected by Toddler signaling. Next, this work will use Western blotting, immunohistochemistry and epistatic analysis to analyze how Toddler regulates E-cadherin, an adhesion molecule known to be active during early development. Finally, this work will examine Toddler's relationship with Apelin, a conserved peptide active in development that, like Toddler, signals through the Apelin Receptors A and B. It is likely that the Toddler and Apelin pathways are partially overlapping, but the degree and requirement for this is unclear. This relationship will be assessed by generation of an apelin mutant followed by single and double mutant analysis including cell tracking and comparisons of morphology and reporter gene expression. As a result, this study will increase our knowledge of cell migration during development thereby enhancing our understanding of how organisms transition from single cells to complex systems. Such knowledge will aid in our ability to understand and address development related diseases. Additionally, the peptide Apelin is known to be involved in cardiovascular development and is currently being tested as a therapeutic in humans and the work carried out in this project may reveal Toddler to be a similarly promising drug target.

描述（由申请人提供）：脊椎动物发育依赖于细胞的精确增殖、迁移和特化，以产生生物体的独特和复杂部分。这些过程中的任何一个失败都可能导致缺陷，如器官衰竭，神经系统综合征或胚胎死亡。尽管这些过程的重要性，他们的途径和监管机构没有完全理解。该项目将通过表征Toddler来解决这一差异，Toddler是一种新型、高度保守的分泌肽，参与胚胎发生，并且是斑马鱼原肠胚形成期间内胚和中胚层正确迁移所需的。实验室先前的工作表明，Toddler调节细胞迁移，尽管Toddler刺激的细胞行为变化和分子机制尚不清楚。这项工作将测试的假设，幼儿是所需的细胞内化原肠胚形成（目标1），由于其调节E-钙粘蛋白（目标2）和部分重叠的功能与已知的分泌肽爱帕琳（目标3）。首先，基因工程和显微镜分析将用于标记和跟踪发育过程中的幼儿反应细胞，以确定受幼儿信号影响的特定细胞行为。接下来，这项工作将使用蛋白质印迹，免疫组织化学和上位性分析来分析幼儿如何调节E-钙粘蛋白，一种已知在早期发育过程中活跃的粘附分子。最后，这项工作将研究幼儿与爱帕琳的关系，爱帕琳是一种在发育中活跃的保守肽，像幼儿一样，通过爱帕琳受体A和B发出信号。很可能幼儿和爱帕琳途径部分重叠，但程度和要求尚不清楚。将通过产生爱帕琳突变体，然后进行单突变体和双突变体分析，包括细胞追踪和形态学与报告基因表达的比较，来评估这种关系。因此，这项研究将增加我们对发育过程中细胞迁移的了解，从而增强我们对生物体如何从单细胞过渡到复杂系统的理解。这些知识将有助于我们了解和应对与发育有关的疾病。此外，已知肽Apelin参与心血管发育，目前正在测试作为人类治疗药物，该项目中进行的工作可能揭示Toddler是一个类似的有前途的药物靶点。