Genetic Tools for the Analysis of Nervous Systems Functions

用于分析神经系统功能的遗传工具

• 批准号: 7869568
• 负责人: HERWIG BAIER
• 金额: $ 12.36万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7869568
• 关键词: Affect; Animals; Architecture; Atlases; Automobile Driving; Behavior; Behavioral; Behavioral Assay; Behavioral Paradigm; Biological Assay; Biomedical Research; Brain; Caenorhabditis elegans; Chemicals; Chloride Channels; Communities; Data; Databases; Enhancers; Fishes; Functional disorder; Genetic; Genetic Techniques; Goals; Human; Image; Individual; Internet; Ivermectin; Label; Larva; Mediating; Morphology; Nervous System Physiology; Nervous system structure; Neuraxis; Neurobiology; Neurons; Pattern; Phenotype; Positioning Attribute; Proteins; Publishing; Reagent; Research; Retinal Ganglion Cells; Shipping; Ships; Structure; System; Techniques; Testing; Tissues; Transgenes; Transgenic Organisms; Visual system structure; Work; Zebrafish; base; cell type; insight; neural circuit; offspring; postsynaptic; presynaptic; programs; promoter; protein function; relating to nervous system; tool

DESCRIPTION (provided by applicant): In this application, we propose to develop genetic tools for the conditional, reversible, and tissue-specific silencing of neuronal activity in zebrafish. We then intend to use these new tools to carry out an enhancer trap screen with the aim of characterizing the zebrafish central nervous system in terms of its connectivity and functional architecture. This program will initially entail the use of the bipartite GAL4/UAS system, driving a chemically gated chloride channel in subsets of neurons. This will allow both spatial control (using tissue specific promoters) and temporal control (based on the presence or absence of the chemical gate) over protein function. A major goal of this proposal is to develop and characterize these new genetic techniques and to make the reagents available to the research community. With the ability to silence neurons conditionally and reversibly, we will then undertake an enhancer trap screen, aiming to identify a variety of expression patterns in the central nervous system. An early goal of this screen will be to characterize the expression patterns in terms of their cellular compositions, the fine structures of their individual neurons, and the pre- and postsynaptic connections that the neurons make. This preliminary work will provide important information about the connectivity within and among distinct structures in the zebrafish brain. With genetic access to these tissues, we will then silence neurons composing the trapped expression patterns, and perform behavioral analyses of the affected fish. By systematically assaying for a variety of behaviors in fish with a wide variety of expression patterns, we will eventually develop a functional atlas of the zebrafish brain, relating neural circuits to the behaviors that they mediate. Various tissues and cell types involved in the same behaviors will likely be connected functionally, and we will investigate such possible relationships anatomically. Health-relatedness: The connectivity of the human and fish nervous systems are very similar. By functionally characterizing the circuitry of the zebrafish brain, we also hope to provide insights into the pathophysiology of human behavior. The reagents and techniques developed under this proposal will advance biomedical research throughout the zebrafish community.

描述（由申请人提供）：在本申请中，我们建议开发遗传工具，用于斑马鱼神经元活动的条件性、可逆性和组织特异性沉默。然后，我们打算使用这些新工具进行增强子陷阱筛选，目的是表征斑马鱼中枢神经系统的连接性和功能架构。该计划最初需要使用二分 GAL4/UAS 系统，驱动神经元子集中的化学门控氯离子通道。这将允许对蛋白质功能进行空间控制（使用组织特异性启动子）和时间控制（基于化学门的存在或不存在）。该提案的一个主要目标是开发和表征这些新的遗传技术，并将这些试剂提供给研究界。凭借有条件和可逆地沉默神经元的能力，我们将进行增强子陷阱筛选，旨在识别中枢神经系统中的各种表达模式。该筛选的早期目标是根据细胞组成、单个神经元的精细结构以及神经元形成的突触前和突触后连接来表征表达模式。这项初步工作将提供有关斑马鱼大脑不同结构内部和之间连接性的重要信息。通过对这些组织的遗传访问，我们将沉默构成被捕获表达模式的神经元，并对受影响的鱼进行行为分析。通过系统地分析具有多种表达模式的鱼类的各种行为，我们最终将开发出斑马鱼大脑的功能图谱，将神经回路与其介导的行为联系起来。参与相同行为的各种组织和细胞类型可能会在功能上相互连接，我们将从解剖学上研究这种可能的关系。健康相关性：人类和鱼类神经系统的连通性非常相似。通过对斑马鱼大脑回路的功能表征，我们还希望能够深入了解人类行为的病理生理学。根据该提案开发的试剂和技术将推动整个斑马鱼界的生物医学研究。