Uncovering links between neuronal transcriptomic and functional profiles in opioid addiction

项目摘要 单细胞转录组学方法的最新进展揭示了一个令人难以置信的多样性， 神经元亚型这对理解物质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， 疾病只发生在活人身上一个关键的未解决的问题是如何将这些联系起来 许多转录组学定义的神经元亚型在药物成瘾中的功能作用 在完整的大脑中在这一建议中，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， 口服阿片类药物自我给药过程中丘脑核中遗传鉴定的细胞类型 对小鼠我们也将采取匡威的方法，使用创新的光学方法来标记 在阿片样物质自我给药的不同阶段活跃的神经元亚群， 然后用一种新的原位测序方法鉴定他们死后的转录组谱， 法我们还描述了扩展这些技术的计划，以便与先进的 体内多光子成像和单细胞转录组学和表观基因组学研究。我们预计这一 该项目将开辟物质使用障碍的新探索途径，并广泛促进 了解阿片类药物中神经元基因调控和功能作用之间的关系 成瘾，这可能会发现新的治疗目标。

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.