Neural Circuits of Reward and Drug-Seeking Behavior in Zebrafish

在美国，估计有210万人对阿片类药物上瘾，阿片类药物被认为是最致命的滥用药物。目前的研究已经在药物滥用的细胞和分子机制方面取得了重大进展，并确定了受成瘾影响的神经底物。然而，要开发改进的治疗方法，需要更深入地了解阿片类药物是如何影响大脑可塑性的。为了研究阿片类药物如何影响大脑可塑性，该项目使用了 斑马鱼与强大的体积(3D)成像方法相结合，包括基于荧光的光片显微镜(LSM)和磁共振成像(MRI)技术以及分子图谱和基因表达分析。这项研究的首要目标是开发一个控制斑马鱼寻求奖励和动机行为的神经系统的网络模型。为了剖析这些电路并了解阿片类药物如何影响大脑的可塑性，该项目的两个目标是将强大的成像技术与行为实验、神经活动读数、轨迹跟踪以及斑马鱼的分子和基因表达分析相结合。 目的1将行为学实验与免疫组织学检测相结合，以磷酸化核糖体蛋白(P)S6为读数，定位成年斑马鱼联合寻找奖赏和成瘾行为过程中的功能回路。激活神经元的分子图谱(荧光陷阱)和RNAseq实验将揭示成瘾状态下基因表达的定量和定性变化。AIM 2结合了几种磁共振成像方法来成像整个大脑电路和 神经化学特征。来自LSM和表观荧光显微镜的组织学数据将被整合，以生成3D分段的MRI图谱，该图谱可用于使用活斑马鱼的功能MRI研究。 该项目的结果将导致一个全面的奖励和决策电路网络模型，这对使用斑马鱼作为成瘾模型至关重要。最终，该项目将确定阿片类药物的使用如何影响组织各级的大脑可塑性。

Within the United States, an estimated 2.1 million people suffer from addiction to opioids, considered the most lethal drugs of abuse. Current research has made significant progress with regard to the cellular and molecular mechanisms of drugs of abuse and identified the neural substrates that are affected by addiction. To develop improved therapeutic treatments, however, requires a deeper understanding of how opioids compromise brain plasticity. To examine how opioids affect brain plasticity, the project uses zebrafish in conjunction with powerful volumetric (3D) imaging approaches including fluorescence-based Light Sheet Microscopy (LSM) and Magnetic Resonance Imaging (MRI) technologies as well as molecular profiling and gene expression analyses. The overarching goal of the study is to develop a network model of those neural systems that control reward-seeking and motivated behaviors in zebrafish. To dissect these circuits and understand how opiods affect the plasticity of the brain, the two aims of the project combine powerful imaging technologies with behavioral experiments, neural activity readouts, tract tracing, and molecular and gene expression analyses in zebrafish. Aim 1 integrates behavioral experiments with immunohistological detection of phosphorylated ribosomal protein (p)S6 as a readout to map functional circuitry during associative reward-seeking and addiction behavior in adult zebrafish. Molecular profiling (phosphor-trap) of activated neurons and RNAseq experiments will reveal quantitative and qualitative gene expression changes during addicted states. Aim 2 combines several magnetic resonance imaging methodologies to image whole brain circuitry and neurochemical characteristics. Histological data derived from both LSM and epi-fluorescence microscopy will be integrated to generate a 3D-segemented MRI atlas that can be used in functional MRI studies employing live zebrafish. The results of the project will lead to a comprehensive network model of reward and decision making circuitry critical for using zebrafish as a model for addiction. Ultimately, the project will identify how opioid use affect brain plasticity across levels of organization.