Brain-spanning and scale-crossing circuitry mediating drive function and dysfunction

这项建议旨在提供电路动力学的快感缺乏，精神症状的理解 具有巨大临床意义的领域，非常适合在实验室动物中进行研究。这项工作， 除了我们最近开发的用于获得全脑细胞分辨率活动读出的方法， 控制，创造了一个强大的和偶然的对齐，使我们能够连接本地和全球神经元 动态，并确定跨脑电路介导的行为驱动器，冲突和决议。 在目标1中，我们确定了不同的完善性基础上的单细胞分辨眶额（OFC）动力学， 行为。我们已经开发出一种时间精确的替代选择老鼠行为模式， 设计与我们的宽视场细胞分辨率成像/记录方法兼容，其中小鼠 在多种动机驱动中进行选择，并根据内部或外部环境调整行动计划。 我们将这种范例沿着我们的细胞分辨率读数和分析，从寻址开始， 在眶额皮层中饥渴难耐我们确定了动机驱动力解决的动力学， 缺乏控制的内部状态，并在存在或不存在外部（社会）背景下，使用我们的新 方法;我们从先前的工作中假设（Jennings等人，《自然》杂志2019年报道，解决这些冲突将 这不仅取决于动物的动机（内部）状态，还取决于外部环境。 在目标2中，我们绘制了动机驱动冲突和解决的因果全球动态图，量化了 这些动机驱动相互作用背后的高速细胞分辨率脑回路动力学 （驱动自然发生;或者，为了利用我们的快速电生理读数，而是在时间上诱导- 在饥饿的情况下，通过光遗传学驱动AGRP神经元，和/或SFO输入到 MnPO在口渴的情况下，使用我们建立的模型和方法;艾伦等人，Science 2019; Jennings et 例如，Nature 2019; Marshel等人，Science 2019）。识别新的区域特定的动力学条件下， 不同的动机驱动和社会背景将反馈给Aim 1成像工作流程， 从我们先前的工作中获得了与社交和口渴驱动相互作用相对应的OFC状态成像的坚实基础。 在目标3中，我们定义了驱动间竞争和相应的全脑动力学的基础细胞。 通过天然活性鉴定的多个单细胞将用我们独特的宽视野靶向光遗传学。 和高分辨率空间光引导技术。我们观察到的细胞系 和因果关系，从相同的细胞详细的三维完整组织（STARmap）转录组学信息 在同一个有机体中。与基于解剖学的解剖学和深层分子数据流的比对允许细胞类型- 解决和单细胞水平的洞察力，并针对，生存驱动竞争和解决过程， 对脑部疾病有着重要的意义和相关性。总之，这里提出的方法将 整合新技术，探索自由移动哺乳动物关键症状域的因果基础。

This proposal is designed to provide circuit-dynamics understanding of anhedonia, a psychiatric symptom domain of enormous clinical significance that is well-suited for study in laboratory animals. This work, alongside our recently-developed methods for obtaining brainwide cellular-resolution activity readout and control, has created a powerful and fortuitous alignment enabling us to bridge local and global neuronal dynamics, and to identify brain-spanning circuitry mediating behavioral drives, conflicts, and resolutions. In Aim 1, we identify single-cell-resolved orbitofrontal (OFC) dynamics underlying distinct consummatory behaviors. We have developed a temporally-precise alternative-choice mouse-behavioral paradigm, crucially designed for compatibility with our wide-field cellular-resolution imaging/recording methods, in which mice select among multiple motivational drives, and adjust action planning in light of internal or external context. We apply this paradigm along with our cellular-resolution readouts and analyses, beginning with addressing both hunger and thirst in OFC. We identify dynamics of motivational drive resolution both in the presence or absence of controlled internal states, and in the presence or absence of external (social) context, using our new methods; we hypothesize from prior work (Jennings et al., Nature 2019) that resolution of these conflicts will depend upon not only the motivational (internal) state of the animal but also the external context. In Aim 2, we map causal global dynamics of motivational drive conflict and resolution, quantifying the high-speed cellular-resolution brainwide circuit dynamics underlying these motivational drive interactions (drives naturally-occurring; or, to leverage our fast electrophysiological readout, instead induced in temporally- precise fashion by optogenetically driving AGRP neurons in the case of hunger, and/or SFO inputs to the MnPO in the case of thirst, using our established models and methods; Allen et al., Science 2019; Jennings et al., Nature 2019; Marshel et al., Science 2019). Identification of novel region-specific dynamics in conditions of varying motivational drive and social context will feed back to inform Aim 1 imaging workflow, already with a firm foundation from our prior work imaging OFC states corresponding to social and thirst drive interaction. In Aim 3, we define cells underlying inter-drive competition and corresponding brainwide dynamics. Multiple single cells identified by natural activity will be optogenetically targeted with our unique wide-field and high-resolution spatial light-guidance technology. We register cellular ensembles observed to be naturally and causally involved, to detailed 3D intact-tissue (STARmap) transcriptomic information from the same cells in the same organism. Alignment with wiring-based anatomy and deep molecular datastreams allow cell-type- resolved and single cell-level insight into, and targeting of, survival drive competition and resolution processes, with both basic significance and relevance to brain disease. Together, the approaches proposed here will integrate novel technology to probe causal underpinnings of key symptom domains in freely-moving mammals.