Discovery and characterization of brain-wide neuromodulatory circuits regulating arousal

项目摘要/摘要 唤醒的内部状态可以显著地影响行为，从感觉处理到认知和 情感功能。唤醒障碍是几种精神障碍的常见症状，包括 抑郁、焦虑、成瘾、注意力缺陷多动障碍和精神分裂症。性唤醒可以改变 在多个时间尺度上，包括在认知功能期间优化表现的快速波动。 较慢形式的觉醒与多种神经调节细胞类型的活动有关，包括那些 释放单胺、乙酰胆碱和许多多肽；相反，快速觉醒主要是 归因于去甲肾上腺素能蓝斑。神经调节剂在行为方面的研究具有挑战性 哺乳动物，因为它们小，深，空间分散，分子多样化；因此，一个 关于快速觉醒背后的神经调节系统还没有进行全面的调查。我建议 通过开发和应用研究幼虫神经调节和唤醒的工具来克服这些障碍 斑马鱼。这些脊椎动物与哺乳动物共享保守的神经调节系统，但它们很小， 透明，所以快速时间尺度唤醒背后的神经调节细胞类型可以在 使用细胞分辨率功能成像的整个大脑的规模。我假设这个倍数 神经调节系统同时起作用，以实现快速的时间尺度唤醒。这项提议的目标是 识别和描述实现唤醒的内部状态的神经调节系统，以及 确定这些系统如何塑造全球神经动力学。在初步的努力中，我开发了一部小说 全脑细胞分辨组织配准方法与活体活动对齐相同神经元 用死后免疫组织化学鉴定多种神经调节细胞类型的记录。在 K99指导阶段，我将使用这种方法来分类与试验相关的神经调节细胞类型 觉醒的试验波动，通过感觉运动反应时间来衡量。我的初步数据显示 多个去甲肾上腺素能、胆碱能、5-羟色胺能、多巴胺能和肽能群体与 唤醒。随后，我将绘制这些与唤醒相关的人群的功能连接性图 通过与我的导师卡尔博士一起训练，在转基因鱼中将全脑成像与光遗传学相结合 戴瑟罗斯和共同导师菲利普·莫兰博士。在R00独立阶段，我将把这些技能应用到 确定觉醒相关神经调节细胞类型对全脑动力学的因果影响以及 唤醒的行为表达。与Deisseroth博士和Mourrain博士在斯坦福大学进行全面培训 大学将为我提供从事与唤醒和其他内部状态相关的研究所需的技能 作为一名独立调查员。这些努力将导致对一类觉醒功能障碍症状的深入了解 在一系列不同的精神障碍中很常见。

Project Summary/Abstract The internal state of arousal can dramatically influence behavior, from sensory processing to cognitive and emotional function. Disrupted arousal is a symptom common to several psychiatric disorders, including depression, anxiety, addiction, attention deficit hyperactivity disorder, and schizophrenia. Arousal can change over multiple timescales, including rapid fluctuations that optimize performance during cognitive functions. Slower forms of arousal are linked to the activity of multiple neuromodulatory cell types, including those releasing monoamines, acetylcholine, and numerous peptides; conversely, rapid arousal has been primarily attributed to the noradrenergic locus coeruleus. Neuromodulators are challenging to investigate in behaving mammals, because they are small, deep, spatially dispersed, and molecularly diverse; consequently, a comprehensive survey of neuromodulatory systems underlying rapid arousal has not been conducted. I propose to overcome these obstacles by developing and applying tools to study neuromodulation and arousal in larval zebrafish. These vertebrates share conserved neuromodulatory systems with mammals, yet are small and transparent, so the neuromodulatory cell types underlying fast-timescale arousal can be exhaustively mapped at the scale of the whole brain using cellular-resolution functional imaging. I hypothesize that multiple neuromodulatory systems act in parallel to implement fast-timescale arousal. The goal of this proposal is to identify and characterize the neuromodulatory systems implementing the internal state of arousal, and determine how these systems shape global neural dynamics. In preliminary efforts, I developed a novel whole-brain cellular-resolution tissue registration method for aligning the same neurons from live activity recordings with postmortem immunohistochemical identification of multiple neuromodulatory cell types. In the K99 mentored phase, I will use this method to catalogue the neuromodulatory cell types correlated with trial-to- trial fluctuations in arousal, measured by sensorimotor reaction times. My preliminary data have revealed multiple noradrenergic, cholinergic, serotonergic, dopaminergic, and peptidergic populations correlated with arousal. I will subsequently map the functional connectivity of these arousal-correlated populations by combining brain-wide imaging with optogenetics in transgenic fish, through training with my mentor Dr. Karl Deisseroth and co-mentor Dr. Philippe Mourrain. In the R00 independent phase, I will apply these skills to determine the causal impact of arousal-correlated neuromodulatory cell types on brain-wide dynamics and the behavioral expression of arousal. Comprehensive training with Dr. Deisseroth and Dr. Mourrain at Stanford University will provide me with the skills required to pursue research related to arousal and other internal states as an independent investigator. These efforts will lead to insights into a class of arousal dysfunction symptoms common to a diverse array of psychiatric disorders.