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

项目总结(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电路动力学- 调节行为，在有回报和风险的情况下。我们应用了代表主要路线的技术 药物滥用研究的机会(光脑成像或OEG，帧投影的独立纤维 光度学或FIP和水凝胶-组织化学，包括星图)，不仅允许观察和 控制基因定义的电路，但同时和独立地控制多个电路元件--在此之前， 在药物改变状态中和之后的行为。在目标1中，我们从最广泛的(全脑)范围开始 唤醒啮齿动物，以不偏不倚的方式识别关键角色和原则，同时保持电路元素- 观察和控制活动的特异性。在技术上，Aim 1的实验将包括光遗传- 针对特定电路和投影以及快速和定量的驱动精确脉冲模式 全脑活动模式的评估。这些不偏不倚的初步全球评估将有力地为 在AIMS 2和AIMS 3中，重点关注更多空间受限的调查，以解决剧烈改变的基本特征 各州。在目标2中，我们在更高的空间和时间分辨率下操作，下一步是详细 急性药物改变状态下因果回路动力学的阐明。因为同样的药物和行为 目标1的条件，现在根据神经元活动水平在个体-动物水平进行定量指导，以 使用我们的人口和特定于预测的记录和干预能力成为目标，我们参与模式 人口和预测活动，以测试对行为的因果影响。在目标3中，我们利用了最高的- 我们新方法的分辨率，在保持地图般的广阔视角的同时实现了单细胞分辨率， 在行为期间和在接触滥用药物期间。对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系综控制和深度转录 对这些单个细胞进行分析。这种对不同神经回路通路的精确观察和控制 与项目2-4中描述的研究方案以及技术和培训核心相互交织。 项目1中的这些实验将共同测试多功能、强大的新电路动力学工具 NIDA中心和更广泛的药物滥用社区，并将应用这些工具来深化我们的 了解急性或慢性改变的药物变化状态，以及将大脑本身作为一个动态系统。

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.