Uncovering links between neuronal transcriptomic and functional profiles in opioid addiction

项目摘要 单细胞转录方法的最新进展揭示了令人难以置信的多样性 神经元亚型。这对理解药物使用障碍提出了挑战。 因为这些亚型只能在死后的脑组织中识别，但物质使用 疾病只发生在活人身上。尚未解决的关键问题之一是如何将这些 多种转录定义的神经元亚型及其在药物成瘾中的功能作用 在完整的大脑中。在这项提案中，我们迈出了弥合这一差距的第一步。首先，我们将使用 创新的光遗传学和电生理学技术记录来自 口服阿片类药物自身给药过程中伏隔核细胞类型的遗传学鉴定 在老鼠身上。我们还将采取相反的方法，使用创新的光学方法来标记 在阿片类药物自我给药的不同阶段活跃的神经元亚群， 然后使用一种新的原位测序技术在死后鉴定它们的转录图谱 方法。我们还描述了扩展这些技术以与高级In 活体多光子成像、单细胞转录和表观基因组研究。我们预料到了这一点 该项目将在药物使用障碍方面开辟新的探索路线，并为 了解神经基因调控与阿片类药物功能作用的关系 上瘾，这可能会确定新的治疗靶点。

Project Summary Recent advances in single-cell transcriptomic methods have revealed an incredible diversity of neuronal subtypes. This presents a challenge for understanding substance use disorders because these subtypes can only be identified in post mortem brain tissue, but substance use disorders occur only in living humans. One of the key unsolved problems is how to link these numerous transcriptomically-defined neuronal subtypes to their functional roles in drug addiction in the intact brain. In this proposal, we take first steps toward bridging this gap. First, we will use innovative optogenetic and electrophysiological techniques to record neuronal activity from genetically identified cell types in the nucleus accumbens during oral opioid self-administration in mice. We will also take the converse approach, using innovative optical methods to label neuronal subpopulations that are active during different phases of opioid self-administration, then identifying their transcriptomic profiles post mortem using a novel in situ sequencing method. We also describe plans to extend these techniques for compatibility with advanced in vivo multiphoton imaging and single-cell transcriptomic and epigenomic studies. We expect this project will open new lines of exploration in substance use disorders and contribute broadly to understanding the relationship between neuronal gene regulation and functional roles in opioid addiction, which may identify new therapeutic targets.