Single-cell whole brain imaging of nicotine intoxication, dependence, and abstinence

摘要 在过去的20年里，神经学家们一直专注于大脑的特定电路操作，以识别 使用生物工程方法控制尼古丁依赖和复发的神经元通路，如 光遗传学和钙成像。这些方法已经确定了两者之间的因果关系 尼古丁对局部神经元的激活和特定的尼古丁相关行为。但是我们仍然有非常多的 由于技术上的差距，几乎不了解大脑作为一个整体是如何处理这些信息的 很难以单细胞分辨率对整个大脑进行成像。这是该领域的一个关键问题，因为尼古丁， 药物和行为治疗影响的是整个大脑，而不仅仅是特定的回路。最近的 即刻早期基因单细胞全脑成像的光片显微镜研究进展 Clear Brains(IDESCO+)使以单细胞分辨率研究全脑功能网络成为可能。 最重要的假设是，长程胆碱能神经元的协调激活与 全脑模块化程度降低，戒断相关行为可部分正常化 FDA批准的药物。该提案的第一个目标是在以下位置确定全脑功能网络 与急性尼古丁中毒、慢性尼古丁依赖、戒断和 使用即刻早期基因的IDESCO+成像技术实现持久戒酒。第二个目标是识别大脑 控制这些状态并预测急性和长期生理和行为的活动模式 使用高级神经网络分析对尼古丁的反应。第三个目标是检验假设 FDA批准的药物将使这些网络变化正常化。这个项目将产生四个结果， 可能会对该领域产生长期的影响：1)识别急性脑损伤的脑功能网络 尼古丁、尼古丁依赖、急性戒断和长期戒断。2)细菌的分子表型 功能网络。3)确定预测戒断相关行为的主要中枢区域。4) FDA批准的治疗烟草使用障碍药物的脑纹特征。5)测试 长程胆碱能神经元协同激活与全脑功能减退有关的理论 模块化和戒断相关行为。

Abstract In the past 20 years, neuroscientists have focused on circuit-specific manipulations of the brain to identify neuronal pathways controlling nicotine dependence and relapse using bioengineering approaches, such as optogenetics and calcium imaging. These approaches have determined a causal relationship between activation of localized neurons by nicotine and specific nicotine-related behaviors. However, we still have very little understanding of how the brain, as a whole, processes this information because of a technical gap making it difficult to image the whole brain at single-cell resolution. This is a critical problem for the field, as nicotine, pharmacological and behavioral treatments affect the brain as a whole and not just specific circuits. The recent development of single-cell whole-brain imaging of immediate-early genes using light-sheet microscopy on cleared brains (iDisco+) has made the study of brain-wide functional networks at single-cell resolution possible. The overarching hypothesis is that coordinated activation of long-range cholinergic neurons is associated with decreased whole-brain modularity and withdrawal-related behaviors that can be partially normalized using FDA-approved medications. The first goal of this proposal is to identify brain-wide functional networks at single-cell resolution associated with acute nicotine intoxication, chronic nicotine dependence, withdrawal, and protracted abstinence using iDisco+ imaging of immediate-early genes. The second goal is to identify the brain activity patterns that control these states and predict the acute and long-term physiological and behavioral response to nicotine using advanced neural network analyses. The third goal is to test the hypothesis that FDA-approved medications will normalize these network changes. This project will lead to four outcomes that will likely produce a long-lasting impact on the field: 1) Identification of the functional brain networks of acute nicotine, nicotine dependence, acute withdrawal, and protracted abstinence. 2) Molecular phenotyping of the functional network. 3) Identification of the major hub regions that predict withdrawal-related behaviors. 4) Characterization of the brainprints of FDA-approved medications for tobacco use disorder. 5) Testing the theory that coordinated activation of long-range cholinergic neurons is associated with decreased whole-brain modularity and withdrawal-related behaviors.