Ultra-fast high-resolution imaging of whole mouse brain for the study of drug addiction

用于药物成瘾研究的小鼠全脑超快高分辨率成像

基本信息

批准号：
10359049
负责人：
ZACHARY FREYBERG
金额：
$ 19.8万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10359049
关键词：
3-Dimensional Acute Address Affect Anatomy Atlases Behavior Behavioral Biological Assay Brain Brain imaging Brain region Cells Chemicals Cocaine Cocaine Abuse Color Complex Corpus striatum structure Development Disease Dopamine Drug Addiction Epidemic Exhibits Fentanyl Fostering Future Genetic Global Change Glutamates Heterogeneity Hour Image Imaging technology Individual Laboratories Laser Scanning Confocal Microscopy Light Maps Medial Midbrain structure Morphologic artifacts Mosaicism Movement Mus Neuroanatomy Neurons Opioid Pharmaceutical Preparations Population Prevalence Process Property Public Health Resolution Rewards Rodent Role Scanning Site Societies Speed System Techniques Technology Time Tyrosine 3-Monooxygenase United States Ventral Tegmental Area Work activity marker addiction base cell type combinatorial conditioned place preference confocal imaging dopamine system dopaminergic neuron drug of abuse drug reinforcement drug reward effective therapy family burden genetic approach high resolution imaging imaging approach neural circuit neuronal patterning neurotransmission new therapeutic target opioid abuse overdose death recombinase regional difference response tool vesicular glutamate transporter 2

项目摘要

PROJECT SUMMARY Opioid and cocaine abuse prevalence has skyrocketed in the United States, fueling the current epidemic of overdose deaths. Despite the public health impact of opioids and cocaine, we still lack a fundamental understanding of the mechanisms by which these drugs work, particularly across cellular and circuit levels. Further understanding of the neuroanatomy of the neural circuitry underlying opioid and cocaine reward is a critical initial step in targeting and elucidating their mechanisms. However, comprehensively visualizing relevant circuits in drug reward has been limited by approaches to contextualize these circuits and their response to drugs of abuse in the whole brain. We developed an approach to rapidly image the whole brain in three-dimensional (3D) space using ultra-fast high-resolution ribbon-scanning confocal microscopy. Our ribbon-scanning confocal imaging approach can image and visualize an entire rodent brain in less than 24 hours, where more conventional approaches (e.g., light-sheet) currently require days or even weeks. Furthermore, our ribbon-scanning confocal approach reaches diffraction-limited resolutions (~200-300nm), enabling us to visualize individual cells in the brain and their ultrastructure. We can apply these unique tools to begin solving the fundamental questions: 1) What is the precise circuitry that defines drug reward? And 2) What are the differential effects of cocaine and opioids on this circuitry? Like many drugs of abuse, cocaine and opioids rely on neurotransmission from dopamine (DA) neurons in the ventral tegmental area (VTA). However, until recently, parsing the connectivity of unique subpopulations of DA neurons and their potential roles in drug reward has been difficult. We developed a suite of intersectional genetic tools to definitively dissect the anatomical and functional properties of these different subpopulations within the same brain. We will integrate our 3D ribbon-scanning confocal imaging of DA neuron subpopulations with immunolabeling of neuronal activity markers to visualize precisely which DA neurons are activated in response to cocaine and opioids. Using whole brain immunolabeling and imaging, we will also visualize and map drug-dependent neuronal activity changes in the whole brain with the potential to reveal new populations of neurons differentially response to cocaine and opioids. Our overall objectives are to: Comprehensively map the distribution of DA neuron subpopulations including DA/glutamate co-transmitting cells relative to the overall DA system within whole brain (Aim 1); and to determine how cocaine and opioids differentially affect the activity of these DA neuron subpopulations (Aim 2). We will generate a comprehensive 3D brain atlas to identify the roles of unique subpopulations of DA neurons highly relevant to cocaine and opioids, which will serve as a proof of principle for the implementation of our ultra-fast high-resolution 3D ribbon-scanning confocal microscopy. Our proposal will foster future development of the first 3D high-resolution comprehensive maps of neurotransmission within in whole brain to study addiction.

项目摘要阿片类药物和可卡因滥用率在美国飙升，加剧了当前的流行病过量的死亡。尽管阿片类药物和可卡因对公共卫生的影响，但我们仍然缺乏基本了解这些药物起作用的机制，尤其是在细胞和电路水平上。对阿片类药物和可卡因奖励的神经回路的神经解剖学的进一步了解是靶向和阐明其机制的关键初步步骤。但是，全面可视化相关药物奖励的电路受到将这些电路及其对毒品的反应进行情境化的方法的限制整个大脑的虐待。我们开发了一种快速在三维中迅速对整个大脑形象的方法（3D）使用超快速的高分辨率丝带扫描共聚焦显微镜空间。我们的丝带扫描共焦成像方法可以在不到24小时内形象和可视化整个啮齿动物的大脑，其中更常规目前需要几天甚至几周的方法（例如，灯页）。此外，我们的丝带扫描共焦方法达到衍射有限的分辨率（〜200-300nm），使我们能够可视化单个细胞大脑及其超微结构。我们可以应用这些独特的工具开始解决基本问题：1）定义药物奖励的确切电路是什么？ 2）可卡因和该电路上的阿片类药物？像许多滥用药物一样，可卡因和阿片类药物都依赖于腹侧盖区域（VTA）的多巴胺（DA）神经元。但是，直到最近，解析 DA神经元及其在药物奖励中的潜在作用的独特亚群很困难。我们开发了一套交叉遗传工具，以确定剖析这些解剖学和功能特性同一大脑中的不同亚群。我们将整合DA的3D胶带扫描共聚焦成像神经元的神经元亚群，对神经元活性标志物进行免疫标记，以准确地可视化哪个神经元响应可卡因和阿片类药物而激活。使用整个大脑免疫标记和成像，我们也将可视化和绘制依赖药物依赖性神经元活性在整个大脑中的变化，有可能揭示新的神经元的种群对可卡因和阿片类药物有差异反应。我们的整体目标是：全面绘制DA神经元亚群的分布，包括DA/谷氨酸共转移细胞相对于整个大脑内的整个DA系统（AIM 1）；并确定可卡因和阿片类药物差异影响这些DA神经元亚群的活性（AIM 2）。我们将产生一个全面的 3D脑图集以确定与可卡因和阿片类药物高度相关的DA神经元的独特亚群的作用，这将作为实施我们的超快速高分辨率3D丝带扫描的原理证明共聚焦显微镜。我们的建议将促进第一个3D高分辨率综合的未来发展整个大脑中神经传递的地图研究成瘾。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Roles of VGLUT2 and Dopamine/Glutamate Co-Transmission in Selective Vulnerability to Dopamine Neurodegeneration.

DOI：
10.1021/acschemneuro.1c00741
发表时间：
2022-01-19
期刊：
ACS CHEMICAL NEUROSCIENCE
影响因子：
5
作者：
Buck, Silas A.;Erickson-Oberg, M. Quincy;Bhatte, Sai H.;McKellar, Chase D.;Ramanathan, Vishan P.;Rubin, Sophie A.;Freyberg, Zachary
通讯作者：
Freyberg, Zachary

Relevance of interactions between dopamine and glutamate neurotransmission in schizophrenia.