Developing Genetic Reagents for the Dissection of Dopaminergic Circuitry

开发用于解剖多巴胺能回路的遗传试剂

基本信息

批准号：
8996213
负责人：
Christopher John Potter
金额：
$ 20.25万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8996213
关键词：
Adult Animal Model Animals Arousal Attention Behavior Biological Neural Networks Brain Caenorhabditis elegans Carboxy-Lyases Cells Communities Complex Computers Coupled Courtship Databases Development Disease Dissection Dopa Dopamine Drosophila genus Drosophila melanogaster Drug Addiction Engineering Enhancers Exhibits Functional disorder Generations Genes Genetic Genetic Enhancer Element Genetic Models Genetic Techniques Genomics Goals Health Human Hyperactive behavior Image Individual Label Lead Learning Link Locomotion Mammals Maps Membrane Memory Mental Depression Mental disorders Methods Modeling Mus Names Neurologic Neuromodulator Neurons Neurophysiology - biologic function Neurotransmitters Other Genetics Parkinson Disease Pattern Reagent Reporter Restless Legs Syndrome Rewards Schizophrenia Signal Transduction Sleep Staging System Techniques Therapeutic Intervention Time Tracer Transgenic Organisms Tyrosine 3-Monooxygenase Work addiction base combinatorial dopamine transporter dopaminergic neuron fly genetic manipulation genetic resource in vivo insight man nervous system disorder neural circuit novel pleasure research study tool

项目摘要

DESCRIPTION (provided by applicant): Dopamine (DA) is a critical neurotransmitter, conserved from C. elegans to man, that regulates a wide variety of behaviors, including learning and memory, courtship behaviors, reward-seeking, and sleep/wake rhythms. Dysfunction of DA neural circuits in turn contributes to disorders such as Parkinson's disease, depression, and drug addiction. Yet how DA neural circuits contribute to these behaviors or disorders is not well-understood. The goal of this work is to develop, characterize, and utilize novel genetic reagents for the dissection of the DA neural network in Drosophila melanogaster. Drosophila is an ideal system to carry out these studies, as fruit flies have only ~250 DA neurons to drive many of the same DA-dependent behaviors found in mammals. Moreover, flies are highly tractable to neural circuit analyses, as they have a short generation time, well- developed genetic techniques and resources, and can be easily maintained in large numbers. In previous work, we generated novel transgenic fly lines that allowed us to manipulate distinct subsets of DA neurons. Using these lines, we identified a single pair of DA neurons that promote arousal by projecting to and directly inhibiting a sleep-promoting circuit. In addition, we have recently developed a novel genetic method, CLAMP (Cell Labeling Across Membrane Partners), which allows for identification, morphological characterization, and functional manipulation of neurons based solely on connectivity patterns. Here, we propose to generate novel transgenic DA driver lines, which will be used for the identification, characterization, and connectivity mapping of the DA neural network in the fly brain. First, we will generate new DA transgenic fly lines, based on the genomic enhancers from different genes that express in DA cells. Second, we will screen established Gal4 lines for expression in subsets of DA cells. By using a combinatorial genetic intersectional approach, these fly lines will collectively generate ~22,600 distinct labeling patterns containing small subsets of DA neurons. These lines will be made available to the scientific community to facilitate functional analyses of the DA neural network. Third, we will create a comprehensive database of DA neurons in the fly brain by 1) identifying and naming individual DA cells and 2) by using computer tracing techniques combined with registration to a standard brain model to label projection patterns. Fourth, by using the CLAMP method to systematically map the connectivity of these DA neurons, we will develop a detailed model of the DA neural network in Drosophila. Understanding how DA circuits in Drosophila function to regulate different behaviors would provide insights into related mechanisms in mammals, including humans, and thus set the stage for circuit-based therapeutic interventions for specific neurological and psychiatric diseases.

描述（由申请人提供）：多巴胺（DA）是一个关键的神经递质，从秀丽隐杆线到人类，包括学习和记忆，求爱行为，寻求奖励和睡眠/唤醒节奏。有助于帕金森氏症，抑郁症和毒品的疾病果蝇只有〜250个DA神经元，可以在哺乳动物中发现许多相同的DA依赖性行为。。开发了一种新型的遗传方法，即跨膜伴侣的细胞标记（细胞标记），它允许仅在苍蝇中的DA神经网络绘制的神经元的鉴定，形态表征和功能操纵。在DA细胞中。蝇脑中的DA神经元的数据库通过1）识别和命名单个DA细胞，以及使用计算机跟踪技术使用注册到标准大脑模型的注册来标记投影模式。 DA神经元，我们将开发果蝇中DA神经网络的详细模型。