Multi-color optical voltage imaging of neural activity in behaving animals

行为动物神经活动的多色光学电压成像

• 批准号: 10166236
• 负责人: MARK J SCHNITZER
• 金额: $ 87.54万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-15 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10166236
• 关键词: Action Potentials; Address; Amino Acid Sequence; Animal Behavior; Animals; Area; BRAIN initiative; Brain; Brain imaging; Categories; Cells; Cephalic; Chronic; Color; Complement; Computer software; Data; Deposition; Devices; Disease; Dorsal; Educational workshop; Electrodes; Engineering; Feedback; Fiber Optics; Fluorescence Resonance Energy Transfer; Genetic; Head; Health; Image; Imaging Device; Imaging Techniques; Individual; Intelligence; Knowledge; Label; Laboratories; Learning; Licensing; Light; Machine Learning; Membrane; Membrane Potentials; Methods; Microscope; Molecular; Monitor; Monoclonal Antibody R24; Mus; Neurons; Opsin; Optical Instrument; Optics; Outcome; Pattern; Performance; Population; Preparation; Proteins; Publishing; Research; Resolution; Resource Sharing; Rodent; Science; Scientist; Shapes; Slice; Specificity; Speed; Surface; Techniques; Testing; Time; Training; Transgenic Organisms; United States National Institutes of Health; Universities; Validation; Variant; Viral Vector; Visit; Work; absorption; awake; brain shape; cell type; design; fly; genetic signature; high throughput screening; imaging facilities; imaging modality; imaging study; improved; information processing; innovation; instrument; machine learning method; microendoscopy; millisecond; mutant; neocortical; neural circuit; neurophysiology; new technology; novel; novel strategies; optogenetics; public repository; relating to nervous system; response; screening; sensor; success; tool; voltage

Abstract Groundbreaking work within the NIH BRAIN Initiative has revealed many new types of neurons and their genetic signatures. The dividends from this research will include sophisticated tools allowing selective genetic access to these cell-types, such as for imaging, optogenetic or tracing studies. To complement these powerful genetic tools, it will be equally important to have new imaging techniques that can reveal how multiple neuron- types work together in the live brain to support information-processing and construct different brain states. To address this challenge, Stanford University and The John B. Pierce Laboratory at Yale University will create optical techniques for imaging the concurrent voltage dynamics of up to 4 separate neuron-types in behaving animals. First, we will combine machine learning methods and an automated, high-throughput protein screening platform to engineer 4 different categories of genetically encoded fluorescent optical indicators of neuronal transmembrane voltage. We will then innovate several types of optical instruments tailored to work in conjunction with the new voltage indicators. These instruments will enable unprecedented studies of voltage rhythms and spiking dynamics in 2–4 genetically identified neuron-types in superficial and deep brain areas of awake behaving animals. One instrument will allow us to track the concurrent, population voltage oscillations of 2 neuron-types in freely behaving rodents. Another instrument, an optical mesoscope, will enable imaging studies of voltage waves and oscillations across the entire neocortical surface of behaving mice. A third device will be a high-speed miniature microscope for tracking neural dynamics at single cell-, single spike-resolution in freely behaving mice. Lastly, we will develop the capability to image with millisecond-scale precision the simultaneous spiking dynamics of 4 targeted neuron-types in either cortical or deep brain areas. Five external beta-tester labs will evaluate all these innovations in live mice and flies and provide critical user-feedback. If our work succeeds, it will be a ‘game-changer’ for studies of brain dynamics, yielding vital knowledge about how different neuron-types synergize their dynamics to shape animal behavior and the brain’s global states in health and disease. To facilitate this outcome, we plan a 5-fold strategy for resource sharing: (i) All voltage-indicator constructs, viral vectors, transgenic flies, software and screening data will be deposited at public repositories for open distribution; (ii) All instrument designs will be published in extensive detail to facilitate replication; (iii) Our novel imaging devices will be integrated into an existing NIH-supported, publicly accessible facility for brain-imaging in rodents; (iv) In project years 2–4, we will conduct 4 training workshops for 40 visiting scientists per year (120 in total) to learn the new technologies firsthand. These visitors will also provide extensive user-feedback; (v) We will license our imaging instruments for commercial distribution. Overall, we expect our project will lead to major conceptual advances in brain science and multiple new technologies that will reshape the practice of mammalian brain imaging.

抽象的 NIH BRAIN Initiative 的开创性工作揭示了许多新型神经元及其 这项研究的好处将包括允许选择性遗传的复杂工具。 获取这些细胞类型，例如用于成像、光遗传学或追踪研究。 遗传工具，拥有新的成像技术同样重要，该技术可以揭示多个神经元如何- 不同类型的大脑在活体大脑中协同工作，支持信息处理并构建不同的大脑状态。 为了应对这一挑战，斯坦福大学和耶鲁大学约翰·B·皮尔斯实验室 将创建光学技术，对多达 4 个独立神经元类型的并发电压动态进行成像 首先，我们将结合机器学习方法和自动化的高通量蛋白质。 筛选平台设计 4 种不同类别的基因编码荧光光学指标 然后，我们将创新几种专门用于神经跨膜电压的光学仪器。 这些仪器与新的电压指示器相结合将实现前所未有的电压研究。 大脑浅层和深层区域 2-4 个基因鉴定的神经元类型的节律和尖峰动态 一种仪器将使我们能够跟踪清醒行为的动物的同时发生的群体电压振荡。 自由行为的啮齿动物的 2 种神经元类型 另一种仪器，即光学介观镜，将能够成像。 研究行为小鼠整个新皮质表面的电压波和振荡。 将是一种高速微型显微镜，用于以单细胞、单尖峰分辨率跟踪神经动力学 最后，我们将开发以毫秒级精度对小鼠进行成像的能力。 皮质或深部大脑区域的 4 种目标神经元类型的同时尖峰动力学。 测试实验室将在活体小鼠和苍蝇中评估所有这些创新，并提供重要的用户反馈。 如果我们的工作成功，它将成为大脑动力学研究的“游戏规则改变者”，产生重要的知识 关于不同神经元类型如何协同其动态来塑造动物行为和大脑的全局 为了促进这一成果，我们规划了五重资源共享战略：(i) 所有 电压指示剂构建体、病毒载体、转基因果蝇、软件和筛选数据将存放在 (ii) 所有仪器设计都将详细公布 促进复制；（iii）我们的新型成像设备将被集成到现有的 NIH 支持的、公开的系统中。 啮齿动物脑成像无障碍设施； (iv) 在项目第 2-4 年，我们将举办 4 次培训研讨会 每年有 40 名访问科学家（总共 120 名）来亲身了解新技术。 提供广泛的用户反馈； (v) 我们将许可我们的成像仪器进行商业分销。 总的来说，我们预计我们的项目将带来脑科学的重大概念进步和多个新的 将重塑哺乳动物脑成像实践的技术。