Cholinergic mechanisms of cocaine reinforcement probed with nicotinic receptor gene editing

项目摘要 滥用障碍是一个重大的公共卫生挑战，迫切需要新的治疗方法。 needed.烟草使用与可卡因使用障碍高度共病，目前尚不清楚 烟碱信号转导是否/如何调节大脑对可卡因的反应。我们假设 烟草的使用有助于可卡因的使用，我们将使用大鼠模型系统来模拟这个研究问题。我们 将探讨尼古丁调节可卡因强化的机制基础，重点是 识别大脑奖赏回路中的关键尼古丁受体和神经细胞类型。在R21阶段， 验证我们在以前资助的项目中开发的一套分子工具。这些工具使我们能够减少 或消除大脑中特定细胞类型中特定尼古丁受体的表达。我们会核实这些 工具在这个项目中发挥了作用，研究它们如何降低细胞中尼古丁受体的功能。 测定（目的1）。这涉及单细胞膜片钳记录和纤维光度测定。在R21目标2中，我们将 确定尼古丁受体功能衰减是否/如何影响可卡因自我给药行为。 影响可卡因强化的具体操作将通过复制研究和其他研究来证实。 然后，对照实验进行到R33阶段以进行进一步研究。在R33中，我们将进一步探索 尼古丁受体调节可卡因强化的机制。R33 Aim 3将探测电路 参与可卡因自我给药的尼古丁受体调节。R33 Aim 4将使用生物物理 一种检验可卡因与尼古丁具有直接急性和长期相互作用的新假设的方法 受体。总之，这些AIM将帮助我们了解尼古丁受体活性如何调节可卡因摄入。 相关的神经生物学机制将被确定，可能导致新的方法来治疗 烟草和/或可卡因依赖

PROJECT SUMMARY Cocaine use disorder represents a major public health challenge, and novel treatment approaches are urgently needed. Tobacco usage is highly co-morbid with cocaine use disorder, and at present, it is not known whether/how nicotinic signal transduction modulates the brain's response to cocaine. We hypothesize that tobacco usage contributes to cocaine use, and we will model this research question using a rat model system. We will probe the mechanistic underpinnings of nicotinic modulation of cocaine reinforcement, focusing on identifying key nicotine receptors and nerve cell types in the brain's reward circuit. In the R21 phase, we will validate a set of molecular tools that we developed in a previously funded project. These tools allow us to reduce or eliminate the expression of specific nicotine receptors in precise cell types in the brain. We'll verify that these tools work for their purpose in this project, examining how well they reduce nicotine receptor function in cellular assays (Aim 1). This involves single-cell patch-clamp recordings and fiber photometry. In R21 Aim 2, we will determine whether/how nicotine receptor functional attenuation impacts cocaine self-administration behavior. Specific manipulations that impact cocaine reinforcement will be confirmed with replication studies and other control experiments then advanced to the R33 phase for further investigation. In the R33, we will further probe the mechanisms by which nicotine receptors modulate cocaine reinforcement. R33 Aim 3 will probe the circuits involved in nicotine receptor modulation of cocaine self-administration. R33 Aim 4 will use a biophysical approach to examine the novel hypothesis that cocaine has direct acute and long-term interactions with nicotine receptors. Together, these AIMs will help us understand how nicotine receptor activity modulates cocaine intake. Associated neurobiological mechanisms will be identified, potentially leading to novel approaches to treat tobacco and/or cocaine dependence.