Cracking the neuromodulation code at single cell resolution

以单细胞分辨率破解神经调节密码

• 批准号: 10223888
• 负责人: Hyungbae Kwon
• 金额: $ 114.63万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-10 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10223888
• 关键词: Animal Behavior; Animals; Behavior; Behavioral; Brain; Cells; Characteristics; Code; Complex; Dopamine; Electrophysiology (science); Emotional; Gene Expression; Image; Individual; Knowledge; Label; Lead; Libraries; Link; Mammals; Mental disorders; Methodology; Modernization; Monitor; Names; Neuromodulator; Neurons; Neurosciences; Pathogenesis; Pathway interactions; Physiological; Population; Property; Reaction; Resolution; Signal Transduction; Slice; Social Behavior; Social Interaction; Structure; Synapses; Techniques; Testing; Time; Vertebral column; Vertebrates; awake; calcium indicator; in vivo; learned behavior; mind control; neuroregulation; novel; optogenetics; real time monitoring; response; spatiotemporal; temporal measurement; two-photon

Project Summary/Abstract One of challenges in modern Neuroscience is to understand circuit mechanisms that lead to complex behaviors. Our ability to monitor neuronal activity in vivo using genetically encoded calcium indicators and various imaging/optogenetic techniques such as two-photon imaging and channelrhodopsin have helped us define real-time changes in neuronal activity and the circuit basis of behaviors. However, learning and behaviors cannot be solely explained by electrophysiological properties of single neurons and their synaptic connectivity because they are modulated by internal brain state. Therefore, we cannot fully understand diverse emotional or behavioral reactions without understanding the internal brain state. Neuromodulators have been suggested as key molecules that control brain state, but their action to neurons has not been understood at cellular resolution. We have recently developed a novel technique (named “iTango2”) that labels and manipulates neuromodulation-sensitive neuronal populations with high spatiotemporal resolution. Using this iTango2 methodology, we would like to dissect neuromodulator circuits at individual cell levels, and their physiological implications related to complex behavior will be explored in this study. In the first aim, we will examine how sparse dopamine projection in the premotor cortex contributes to cortical circuit assembly, which may uncover cellular mechanisms of the asymmetric principle of cortical neuronal connectivity. Second, we will dissect neuromodulation signaling at subcellular resolution. This will be accomplished by creating a synapse version of iTango2, “Syn-iTango2”. In order to identify potential cortical layer- or dendritic branch-specific mechanisms, we will perform real-time monitoring of local dendritic activation triggered by neuromodulatory inputs in brain slices as well as awake behaving animals, and concomitant structural changes such as spine formation or enlargement will be examined. In the third aim, we would like to identify the neuronal ensemble responsible for social interaction, one of the essential complex behaviors in mammals. Since iTango2 links neuromodulation signals to gene expression, we will test the sufficiency of identified circuits to social behavior. Last, we will build a large library of iTango2, so that this approach becomes broadly useful to a variety of neuroscientists. In summary, completion of this study will demonstrate fine scale of neuromodulation action in a quantitative manner rather than a simple “ON” or “OFF” effect of neuromodulation. Monitoring neuromodulatory effects and manipulating populations of cells with high spatial and temporal resolution would dramatically increase our knowledge of the pathways that underlie vertebrate animal behaviors.

项目总结/摘要 现代神经科学的挑战之一是了解导致神经元死亡的电路机制。 复杂的行为我们利用基因编码技术在体内监测神经元活动的能力 钙指示剂和各种成像/光遗传学技术，如双光子成像， 通道视紫红质已经帮助我们确定了神经元活动和回路的实时变化 行为的基础。然而，学习和行为不能仅仅用 单个神经元的电生理特性及其突触连接，因为它们 是由大脑内部状态调节的因此，我们不能完全理解不同的情感， 或行为反应而不了解内部大脑状态。 神经调质被认为是控制大脑状态的关键分子，但它们的 对神经元的作用还没有在细胞分辨率上被理解。我们最近开发了一种 一种新的技术（名为“iTango 2”），标记和操纵神经调节敏感的 具有高时空分辨率的神经元群体。使用iTango 2方法，我们 我想在单个细胞水平上剖析神经调质回路， 本研究将探讨与复杂行为相关的影响。在第一个目标中，我们将 研究前运动皮层中稀疏的多巴胺投射如何影响皮层回路 组装，这可能揭示皮层神经元不对称原理的细胞机制， 神经元连接其次，我们将在亚细胞水平上剖析神经调节信号， 分辨率这将通过创建iTango 2的突触版本“Syn-iTango 2”来实现。 为了确定潜在的皮质层或树突分支特异性机制，我们将 实时监测由神经调节输入触发的局部树突激活， 大脑切片以及清醒行为的动物，以及伴随的结构变化， 将检查脊柱形成或扩大。在第三个目标中，我们希望确定 负责社会互动的神经元系综，是人类社会中重要的复杂行为之一， 哺乳动物由于iTango 2将神经调节信号与基因表达联系起来，我们将测试iTango 2的基因表达。 社会行为的识别回路的充分性。最后，我们将建立一个大型的iTango 2库， 因此，这种方法对各种神经科学家都很有用。 总之，本研究的完成将证明神经调节作用的精细程度， 定量方式而不是神经调节的简单“开”或“关”效应。监测 神经调节作用和操纵具有高空间和时间的细胞群 分辨率将极大地增加我们对脊椎动物的潜在途径的了解， 动物行为。