G-Protein Coupled Receptors and Axon Guidance in the Drosophila Embryonic Nerve C

果蝇胚胎神经 C 中的 G 蛋白偶联受体和轴突引导

• 批准号: 8051863
• 负责人: MARK F VANBERKUM
• 金额: $ 7.06万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8051863
• 关键词: Address; Affect; Arrestins; Axon; Behavioral; Biological Assay; Biological Models; Cells; Cerebellum; Complex; Congenital Abnormality; Cues; Cyclic AMP; Data; Defect; Development; Developmental Process; Drosophila genus; Embryo; Embryonic Nervous System; Endocytosis; Event; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Genes; Genetic; Genetic Techniques; Goals; Height; Hippocampus (Brain); Homologous Gene; Human Development; In Situ Hybridization; Individual; Laboratories; Lead; Life Style; Ligands; Link; Membrane; Methods; Microarray Analysis; Mission; Modeling; Molecular; Molecular Genetics; Mutation; Nerve; Nervous system structure; Neurologic; Operating System; Optic tract structure; Pathology; Pathway interactions; Persons; Process; Psyche structure; Quality of life; RNA; Research Personnel; Role; Second Messenger Systems; Signal Pathway; Signal Transduction; Specificity; Spinal Cord; Spinal Ganglia; Staging; Techniques; Testing; Therapeutic Intervention; United States National Institutes of Health; axon guidance; base; behavioral impairment; extracellular; gene conservation; gene function; genetic analysis; improved; in vivo; nervous system development; receptor; receptor function; scaffold; second messenger; tool

DESCRIPTION (Provided by Applicant): The development of axon pathways is a complex signal transduction process linking extracellular guidance cues to intracellular signals regulating axon outgrowth and steering. Guidance errors lead to structural birth defects in the nervous system that cause physical, mental, or behavioral impairment. The long-term goal of the investigator's laboratory is to understand how signaling events lead to the development of axon pathways and functional connectivity. Drosophila will be used as the model system as it allows molecular, genetic, and cellular tools to be applied to the in vivo study of axon guidance, while gene conservation makes the study relevant to human development. The goal of this application is to establish a role for G-protein coupled receptors (GPCRs) in axon pathway formation during development of the Drosophila embryonic nervous system. GPCRs are transmembrane receptors that activate trimeric G-proteins to regulate several key intracellular signaling pathways. GPCRs may directly function as guidance cue receptors, or modify second messenger levels to indirectly affect the signaling potential of other ligand-receptor systems operating at a guidance choice point. Based on vertebrate and Drosophila studies, it is hypothesized that GPCRs function in the developing Drosophila nerve cord to guide axons to their targets. Specific Aim 1 will test this hypothesis by systematically evaluate the role of GPCRs in axon pathway formation in the Drosophila embryo by genetically altering components of the canonical GPCR signaling pathway; and Specific Aim 2 will identify GPCRs that guide the development of axon pathways. Aim 1 tests how mutations in various genes function in GPCR signaling pathways alter axon pathway formation alone or in combination with other guidance molecules. It documents the importance of GPCR signaling to pathway formation and addresses how they do so. Aim 2 uses standard RNA hybridization methods to identify GPCRs expressed in the nerve cord. New preliminary data have led the investigator to focus on Methuselah and its Methuselah-like cousins, two of which are expressed in the nerve cord during axonogenesis. Mutations in these genes are used to assess the importance of expression to axon guidance. Future R01 applications will harness the Drosophila "toolbox" to delineate the molecular mechanism by which a GPCR participates in axon guidance decisions. Understanding the fundamental processes underlying axon pathway formation, and their association with structural birth defects, is a critical component of the NIH mission. Elucidating the mechanisms of GPCR activity also contributes to our understanding of several neurological and behavioral pathologies. PROJECT NARRATVE: Errors in the development of axon pathways lead to structural birth defects, which in turn lead to physical, mental, and behavioral impairments significantly impacting a person's lifestyle and the national economy. This application uses the development of the Drosophila embryo as a model system to understand the fundamental processes underlying development of axon pathways in the spinal cord. Understanding these processes is an important step in devising therapeutic interventions and improving the quality of life of affected individuals.

描述（由申请人提供）：轴突通路的发育是一个复杂的信号转导过程，将细胞外引导信号与调节轴突生长和转向的细胞内信号联系起来。 指导错误会导致神经系统结构性先天缺陷，从而导致身体、精神或行为障碍。 研究人员实验室的长期目标是了解信号事件如何导致轴突通路和功能连接的发展。 果蝇将被用作模型系统，因为它允许将分子、遗传和细胞工具应用于轴突引导的体内研究，而基因保守使该研究与人类发育相关。 本申请的目的是确定 G 蛋白偶联受体 (GPCR) 在果蝇胚胎神经系统发育过程中轴突通路形成中的作用。 GPCR 是跨膜受体，可激活三聚体 G 蛋白以调节多个关键的细胞内信号传导途径。 GPCR 可以直接充当引导信号受体，或修改第二信使水平以间接影响在引导选择点运行的其他配体-受体系统的信号传导潜力。 基于脊椎动物和果蝇研究，假设 GPCR 在发育中的果蝇神经索中发挥作用，引导轴突到达其目标。 具体目标 1 将通过遗传改变经典 GPCR 信号通路的组成部分，系统地评估 GPCR 在果蝇胚胎轴突通路形成中的作用，从而检验这一假设；具体目标 2 将确定指导轴突通路发育的 GPCR。 目标 1 测试 GPCR 信号通路中各种基因的突变如何单独或与其他引导分子结合改变轴突通路的形成。 它记录了 GPCR 信号传导对通路形成的重要性，并阐述了它们如何做到这一点。 目标 2 使用标准 RNA 杂交方法来识别神经索中表达的 GPCR。 新的初步数据使研究人员将注意力集中在玛土撒拉及其类似玛土撒拉的表亲上，其中两种在轴突发生过程中在神经索中表达。 这些基因的突变用于评估表达对轴突引导的重要性。 未来的 R01 应用将利用果蝇“工具箱”来描述 GPCR 参与轴突引导决策的分子机制。 了解轴突通路形成的基本过程及其与结构性出生缺陷的关联是 NIH 使命的重要组成部分。 阐明 GPCR 活性的机制也有助于我们理解几种神经和行为病理学。 项目叙述：轴突通路发育错误会导致结构性出生缺陷，进而导致身体、精神和行为障碍，严重影响一个人的生活方式和国民经济。 该应用程序使用果蝇胚胎的发育作为模型系统来了解脊髓中轴突通路发育的基本过程。 了解这些过程是设计治疗干预措施和改善受影响个体生活质量的重要一步。