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

这项提案旨在提供对快感缺失的电路动力学理解，快感缺失是一种精神症状。 具有巨大临床意义的领域，非常适合在实验室动物身上进行研究。这部作品， 除了我们最近开发的获得全脑细胞分辨率活动读数和 控制，创造了一种强大而偶然的排列，使我们能够连接局部和全局神经元 动力学，并确定调节行为驱动、冲突和解决方案的跨脑电路。 在目标1中，我们确定了支持不同炎症的单细胞分辨的眼眶前额(OFC)动力学。 行为。我们已经开发出了一种时间上精确的替代选择老鼠行为范式，关键是 专为与我们的广域细胞分辨率成像/记录方法兼容而设计，在这些方法中，小鼠 从多种激励动力中进行选择，并根据内部或外部环境调整行动计划。 我们将这一范例与我们的细胞分辨率读数和分析一起应用，从寻址开始 在OFC中既饥饿又口渴。我们识别动机驱动力解决的动态，无论是在存在还是 没有受控的内部状态，以及在存在或不存在外部(社会)上下文的情况下，使用我们的新 方法；我们根据先前的工作(Jennings等人，《自然》，2019年)假设这些冲突的解决将 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全脑电路动力学 (驱动是自然发生的；或者，利用我们快速的电生理读数，而不是在时间上感应- 在饥饿的情况下，通过光遗传驱动AGRP神经元和/或SFO输入到 在口渴的情况下，使用我们建立的模型和方法；Allen等人，《科学》2019；Jennings等人 等人，《自然》2019；Marshel等人，《科学》2019)。识别新的区域特定动力学在以下条件下 不同的动机和社交背景将反馈给Aim 1成像工作流程，该工作流程已经具有 我们之前的工作奠定了坚实的基础，想象出与社交和口渴驱动相互作用相对应的OFC状态。 在目标3中，我们定义了潜在的驱动间竞争的细胞和相应的全脑动力学。 通过自然活动确定的多个单细胞将通过我们独特的宽视场进行光遗传靶向 和高分辨率空间光导技术。我们登记了观察到的自然细胞群 和因果相关，到来自相同细胞的详细3D完整组织(星图)转录信息 在同一生物体中。与基于有线的解剖和深分子数据流的比对允许细胞类型- 已解决的和单一细胞级别的对生存的洞察和目标，推动竞争和解决过程， 既具有基本意义，又与脑部疾病相关。总之，这里提出的方法将 整合新技术，探索自由活动的哺乳动物关键症状领域的因果基础。

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.