Single-cell causality in origination, propagation, and resolution of drug-altered brain states

项目概要（项目1） 在这里，我们实施了一种技术驱动的方法来识别和控制潜在的药物回路动力学， 调节行为，在带有奖励和风险的环境中。我们应用的技术代表了一个主要的对齐 药物滥用研究的机会（脑电图或OEG，框架投射独立纤维 光度法或FIP以及水凝胶组织化学（包括STARmap），不仅可以进行观察， 控制遗传定义的电路，但同时和独立的多个电路元件-之前， 在药物改变状态下的行为期间和之后。在目标1中，我们开始在最广泛的（全脑）规模， 清醒的啮齿动物，以公正的方式确定关键球员和原则，同时保持电路元件- 观察和控制活性的特异性。从技术上讲，目标1实验将包括光遗传学- 驱动针对特定电路和投影的精确脉冲模式， 全脑活动模式的评估。这些初步的公正的全球评估将有力地提供信息， 在目标2和目标3中集中更多的空间限制调查，以解决急性改变的基本特征， states.在目标2中，我们以更高的空间和时间分辨率进行操作，下一步是详细的 阐明急性药物改变状态下的因果回路动力学。同样的药物和行为 目标1中的条件，现在在个体动物水平上就神经元活动水平进行定量指导， 使用我们的人口和项目特定的记录和干预能力，我们发挥在模式 人口和投射活动来测试对行为的因果影响。在目标3中，我们利用最高的- 我们的新方法的分辨率，实现单细胞分辨率，同时保持地图般的广阔前景， 在行为和暴露于滥用药物的过程中。因果关系的干预性检验将直接指导 通过这些观察，并将建立在单细胞分辨率集合控制和深层转录组学基础上， 分析这些细胞。这种对不同神经回路通路的精确观察和控制， 与项目2-4中描述的研究计划以及技术和培训核心交织在一起。 总之，项目1中的这些实验将测试多功能，功能强大的新电路动力学工具，用于 NIDA中心和更广泛的药物滥用社区，并将应用这些工具，以深化我们的 理解急性或慢性改变的药物改变状态，以及大脑本身作为一个动力系统。

PROJECT SUMMARY (Project 1) Here we implement a technology-driven approach to identify and control circuit dynamics underlying drug- modulated behaviors, in contexts carrying reward and risk. We apply technology representing a major alignment of opportunity for drug abuse research (optoencephalography or OEG, frame-projected independent-fiber photometry or FIP, and hydrogel-tissue chemistry including STARmap) which allow not only observation and control of genetically-defined circuitry, but multiple circuit elements simultaneously and independently-- before, during and after behaviors in the drug-altered state. In Aim 1, we begin at the broadest (brainwide) scale in awake rodents, to identify key players and principles in unbiased fashion, while maintaining circuit element- specificity for observation and control of activity. Technologically, Aim 1 experiments will include optogenetically- driven precise pulse patterns targeted to specific circuits and projections, as well as rapid and quantitative assessment of brainwide activity patterns. These initial unbiased global assessments will powerfully inform and focus more spatially-restricted investigations in Aims 2 and 3 that resolve essential features of acutely altered states. In Aim 2 we operate at much higher spatial and temporal resolution, the next step toward detailed elucidation of causal circuit dynamics in the acutely drug-altered state. For the same drug and behavioral conditions in Aim 1, now quantitatively guided at the individual-animal level in terms of neuronal activity levels to be targeted using our population and projection-specific recording and intervention capability, we play-in patterns of population and projection activity to test causal impact on behavior. And in Aim 3, we leverage the highest- resolution of our new methods, that achieve single-cell resolution while maintaining map-like broad perspective, during behavior and during exposure to drugs of abuse. Interventional tests for causality will be directly guided by these observations, and will build upon both single-cell-resolution ensemble control and deep transcriptomic analysis of those individual cells. This precise observation and control of distinct neural circuit pathways is tightly intertwined with the research programs described in Projects 2-4, and the Technology and Training Cores. Together, these experiments in Project 1 will test versatile, powerful new circuit-dynamics tools for use in the NIDA Center and for the drug abuse community more broadly, and will also apply these tools to deepen our understanding of acute or chronically-altered drug altered states, and of the brain itself as a dynamical system.