Delineating the genetic basis of amphetamine sensitivity using a Drosophila behavioral model

项目摘要 滥用精神兴奋剂，包括安非他明（AMPH），是一个重大的公共卫生问题， 精神、医疗和心理社会并发症。遗传因素在很大程度上决定了一个人的 然而，对风险等位基因的研究取得了有限的成功。的 对精神兴奋剂的初始敏感性差异很大，并与持续使用和 虐待这种特性可以在动物模型中进行研究，动物模型已经成为研究这种疾病的有力工具。 对药物的行为反应是有控制的和系统的。各种方法的结合， 在这个应用程序中提出的一个建议将使我们能够利用果蝇遗传学的力量来发现新的基因 和赋予对AMPH敏感性的基因变异。由于其快速的生命周期和遗传、细胞 和分子分析，苍蝇使行为研究领域，更难以调查 在脊椎动物模型中。本申请中提供的初步数据表明，苍蝇对 通过增加他们的运动活动和减少他们的睡眠。基因突变会破坏 多巴胺（DA）合成或多巴胺转运蛋白基因（DAT）功能抑制这些行为反应， 这表明我们已经开发出一种强大的行为工具，将遗传变异与表型相关联， 变化我们已经制定了一项战略，将这种行为分析与下一代（Next-Gen） 测序技术和系统遗传学方法研究AMPH的遗传结构 敏感性，并确定新的基因变异，影响这一特点。这一综合办法的实现是因为 我们与神经遗传学实验室的大卫戈德曼博士和科林霍奇金森博士的积极合作 在NIAAA，专家在国家的最先进的下一代技术，遗传连锁研究和功能基因组学 行为特征的研究方法。我们提出以下具体目标：1）鉴定基因 在野生型AMPH的亚株内和亚株之间， 型，非等基因果蝇株Canton S（CS）。我们将（a）使用选择性育种，结合 基因组方法，如全基因组测序和缺陷图谱，以确定基因变异， 改变不同CS亚株中的AMPH敏感性，和B）使用RNA测序（RNA-seq）来分析基因 在不同的亚株中与AMPH敏感性改变相关的表达变化和2）筛选一个 大群体的近交系，以确定与AMPH敏感性改变相关的遗传位点。 我们将（a）筛选果蝇遗传参考组，其中包括203个基因型近交系， 对AMPH的反应和（B）使用系统遗传学方法，包括数量性状基因座（QTL）分析 和极端QTL定位，关联表型变异的敏感性与DNA多态性的AMPH。

Project Summary Abuse of psychostimulants, including amphetamines (AMPHs), is a major public health problem with profound psychiatric, medical and psychosocial complications. Genetic factors contribute substantially to an individual's susceptibility to developing addiction; however, the search for risk alleles has yielded limited success. The initial sensitivity to psychostimulants varies significantly, and has been associated with continued use and abuse. This trait can be studied in animal models, which have emerged as powerful tools to investigate the behavioral response to drugs in a controlled and systematic manner. The combination of approaches we propose in this application will allow us to harness the power of Drosophila genetics to uncover novel genes and gene variants that confer sensitivity to AMPH. With its rapid life cycle and accessibility to genetic, cellular and molecular analyses, the fly has enabled behavioral studies in areas that are far more difficult to investigate in vertebrate animal models. The preliminary data presented in this application show that that flies respond to AMPH by increasing their locomotor activity and decreasing their sleep. Genetic mutations that disrupt dopamine (DA) synthesis or dopamine transporter gene (DAT) function inhibit these behavioral responses, demonstrating that we have developed a robust behavioral tool to associate genetic variations with phenotypic changes. We have developed a strategy combining this behavioral analysis with next-generation (Next-Gen) sequencing technology and systems genetics approaches to investigate the genetic architecture of AMPH sensitivity and identify new gene variants that influence this trait. This integrated approach is made possible by our active collaboration with Dr. David Goldman and Dr. Colin Hodgkinson at the Laboratory of Neurogenetics at NIAAA, experts in state-of-the-art Next-Gen technologies, genetic linkage studies and functional genomics approaches to the study of behavioral traits. We propose the following specific aims: 1) To identify gene variants that underlie wide variation in AMPH sensitivity within and between substrains of the wild- type, non-isogenic Drosophila strain Canton S (CS). We will (a) use selective breeding, combined with genomic approaches such as whole genome sequencing and deficiency mapping, to identify gene variants that alter AMPH sensitivity in different CS substrains and b) use RNA-sequencing (RNA-seq) to profile gene expression changes associated with altered sensitivity to AMPH in the different substrains and 2) To screen a large population of inbred strains to identify genetic loci associated with altered sensitivity to AMPH. We will (a) screen the Drosophila Genetic Reference Panel, which consists of 203 genotyped inbred lines, for response to AMPH and (b) use systems genetics approaches, including quantitative trait locus (QTL) analysis and extreme QTL mapping, to associate phenotypic variation in sensitivity to AMPH with DNA polymorphisms.