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: 7661421
• 负责人: HERWIG BAIER
• 金额: $ 30.9万
• 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/7661421
• 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，我们建议开发斑马鱼中神经元活性的条件，可逆和组织特异性沉默的遗传工具。然后，我们打算使用这些新工具来执行增强器陷阱屏幕，目的是在其连接性和功能架构方面表征斑马鱼中央神经系统。该程序最初将需要使用两分的GAL4/UAS系统，从而在神经元子集中驱动化学门控氯化物通道。这将允许空间对照（使用组织特异性启动子）和时间对照（基于化学栅极的存在）在蛋白质功能上。该提案的主要目的是开发和表征这些新的遗传技术，并使研究界可用的试剂可用。凭借有条件和可逆的神经元沉默的能力，我们将进行增强子陷阱屏幕，旨在识别中枢神经系统中的各种表达模式。该屏幕的一个早期目标是根据其细胞组成，单个神经元的细胞结构以及神经元产生的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内部和不同结构之间的连通性的重要信息。随着对这些组织的遗传获取，我们将使组成被捕获的表达模式的神经元沉默，并对受影响的鱼进行行为分析。通过系统地分析具有各种表达模式的鱼类的各种行为，我们最终将开发斑马鱼大脑的功能性地图集，将神经回路与它们介导的行为有关。涉及相同行为的各种组织和细胞类型可能会通过功能连接，我们将在解剖学上研究这种可能的关系。与健康相关性：人类和鱼神经系统的连通性非常相似。通过功能表征斑马鱼大脑的电路，我们还希望提供有关人类行为病理生理学的见解。根据该提案开发的试剂和技术将推进整个斑马鱼社区的生物医学研究。