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: 10488632
• 负责人: Geoffrey A Dilly
• 金额: $ 3.7万
• 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 2023-09-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10488632
• 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.

项目概要/摘要： 研究：酒精使用障碍是非常普遍和昂贵的，只有少数适度有效 药物治疗是可用的。目前的证据表明，反复饮酒 诱导中央杏仁核（CeA）神经元内基因表达的变化， 酒精依赖，这些变化驱动病理性酒精消费。例如，药理学 研究表明，反复的酒精暴露会增加促肾上腺皮质激素释放因子（CRF）信号， CeA，它促进啮齿动物过度饮酒。然而，本CRF的来源并非 这是肯定的，也不太可能增加CRF信号是增加酒精消费的唯一驱动力 在这些动物身上。这个项目将测试的假设，外侧CeA内的神经元的来源， CRF和这些神经元产生额外的神经肽，驱动过量饮酒。这 将通过检查个体下调后的饮酒行为来检验这一假设。 使用BAC转基因Crh-Cre大鼠的CeA-CRF神经元中的神经递质CRF、强啡肽或GABA， 依赖Cre的RNA干扰该项目还将使用无偏见的方法来探讨酒精引起的 通过使用单个核RNA-seq鉴定神经元群体， 这些种群中受影响最大的基因会成为酒精依赖者。结果 这项工作将导致新的，可测试的假设，特定的蛋白质，可以有针对性地减少 酒精依赖者过度饮酒。 培训：本研究将对申请人进行几项实验技术培训，包括啮齿动物饮酒 行为，遗传工具的测试和验证，单核测序，RNA定量，以及 生物信息学分析工具。通过这项培训，申请人将发展在使用分子生物学方面的专业知识。 操纵基因表达的工具和分析转录组的生物信息学方法。培训 该计划还将培养专业技能，包括科学写作和演示，以及编程， 进一步实现申请人成为独立学术神经科学家的目标。