Cell-type specific central amygdala neurotransmission in alcohol dependence

酒精依赖中细胞类型特异性中央杏仁核神经传递

• 批准号: 10315123
• 负责人: Geoffrey A Dilly
• 金额: $ 3.66万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-07 至 2023-09-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10315123
• 关键词: Acute; Affect; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohols; Algorithms; Amygdaloid structure; Animals; Bacterial Artificial Chromosomes; Behavior; Bioinformatics; Brain region; CRF receptor type 1; Cell Nucleus; Cells; Chronic; Complex; Corticotropin-Releasing Hormone; Drug usage; Dynorphins; Ethanol; Future; Gene Expression; Genes; Genetic; Goals; Heavy Drinking; Human; Individual; Lateral; Lead; Modeling; Modernization; Molecular; Neurons; Neuropeptides; Neurotransmitters; Pathologic; Pharmacological Treatment; Pharmacology Study; Population; Production; Proteins; RNA; RNA Interference; Rattus; Research Training; Rodent; Role; Sampling; Signal Transduction; Small Nuclear RNA; Source; Stress; Techniques; Technology; Testing; Time; Tissues; Training; Training Programs; Transcript; Transgenic Organisms; Validation; Withdrawal; Work; Writing; alcohol abuse therapy; alcohol effect; alcohol exposure; alcohol measurement; alcohol relapse; alcohol research; alcohol use disorder; behavior test; behavioral response; cell type; cost; differential expression; drinking; drinking behavior; emotional stimulus; experimental study; gamma-Aminobutyric Acid; knock-down; mRNA sequencing; neurotransmission; neurotransmitter release; next generation sequencing; optogenetics; prodynorphin; release factor; skills; small hairpin RNA; small molecule; tool; transcriptome; transcriptome sequencing; vapor

Project Summary/Abstract: Research: Alcohol use disorder is highly prevalent and costly, and only a few modestly effective pharmacological treatments are available. Current evidence indicates that repeated consumption of alcohol induces changes in gene expression within neurons in the central amygdala (CeA), and as animals become alcohol dependent, these changes drive pathological alcohol consumption. For example, pharmacological studies have shown that repeated alcohol exposure increases corticotropin-releasing factor (CRF) signaling in the CeA, which promotes excessive alcohol consumption in rodents. However, the source of this CRF is not certain and it is also unlikely that increased CRF signaling is the sole driver of increased alcohol consumption in these animals. This project will test the hypothesis that neurons within the lateral CeA are the source of this CRF and that these neurons produce additional neuropeptides that drive excessive alcohol drinking. This hypothesis will be tested by examining drinking behavior after individually downregulating the neurotransmitters CRF, dynorphin, or GABA in CeA-CRF neurons using a BAC transgenic Crh-Cre rat and Cre-dependent RNA interference. The project will also use an unbiased approach to explore alcohol-induced gene expression in all neurons of the CeA by using single nuclei RNA-seq to identify neuronal populations and the genes within these populations that are most affected as animals become alcohol dependent. The results of this work will lead to new, testable hypotheses about specific proteins that could be targeted to reduce excessive drinking in alcohol dependent individuals. Training: This research will train the applicant in several experimental techniques including rodent drinking behavior, testing and validation of genetic tools, single-nuclei sequencing, RNA quantification, and use of bioinformatics analysis tools. Through this training, the applicant will develop expertise in the use of molecular tools to manipulate gene expression and in bioinformatics approaches to analyze transcriptomes. The training program will also develop professional skills, including scientific writing and presentation, and programming, to further the applicant’s goal of becoming an independent academic neuroscientist.

项目摘要/摘要： 研究：饮酒障碍高度普遍且昂贵，只有少数几乎是有效的 提供药理治疗。当前的证据表明饮酒反复消费 诱导中央杏仁核（CEA）神经元内基因表达的变化，并随着动物的变化 酒精依赖于这些变化促进病理饮酒。例如，药物 研究表明，重复的酒精暴露增加了促肾上腺素释放因子（CRF）信号传导 CEA可促进啮齿动物的过量饮酒。但是，此CRF的来源不是 确定CRF信号的增加是饮酒量增加的唯一驱动力，也不太可能 在这些动物中。该项目将检验以下假设：侧CEA内的神经元是其中的根源 CRF并且这些神经元产生额外的神经肽，以驱动多余的饮酒。这 假设将在单独下调后检查饮酒行为来检验 使用BAC转基因CRH-CRE大鼠和 CRE依赖性RNA干扰。该项目还将采用公正的方法来探索酒精引起的 通过使用单核RNA-Seq鉴定神经元群体和CEA的所有神经元中的基因表达 这些人群中的基因受到动物的依赖而受到影响的最大影响。结果 这项工作将导致有关特定蛋白质的新的，可检验的假设 依赖酒精的人过量饮酒。 培训：这项研究将培训申请人的几种实验技术，包括啮齿饮用 遗传工具的行为，测试和验证，单核测序，RNA定量和使用 生物信息学分析工具。通过这项培训，申请人将在使用分子方面发展专业知识 操纵基因表达和生物信息学方法的工具以分析转录组。培训 计划还将发展专业技能，包括科学写作和演讲以及编程 进一步，申请人成为独立的学术神经科学家的目标。