High-throughput approaches to local and long-range synaptic connectivity

局部和远程突触连接的高通量方法

• 批准号: 10025780
• 负责人: Edward S. Boyden
• 金额: $ 332.57万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10025780
• 关键词: Address; Anatomy; Animal Behavior; Area; Auditory area; Axon; Bar Codes; Brain; Cognition; Communities; Complex; Consumption; DNA; Detection; Disease; Distant; Electron Microscopy; Equipment; Fiber; Functional disorder; Future; Genetic; Health; Immunolabeling Technics; In Situ; Individual; Label; Laboratories; Learning; Link; Location; Maps; Microscopy; Molecular; Mus; Neurons; Neuropil; Neurosciences; Neurosciences Research; Optical Methods; Pathologic; Pathway interactions; Pattern; Problem Solving; Process; Proteins; RNA; Resolution; Specimen; Synapses; Synaptic Cleft; Techniques; Technology; Time; Tissues; Travel; Visible Radiation; base; brain circuitry; connectome; cost; fluorophore; in situ sequencing; light microscopy; millimeter; neural circuit; neuropsychiatric disorder; neuropsychiatry; next generation; novel; relating to nervous system; skills; success; tool

Project Summary/Abstract The overarching objective of this proposal is to develop a robust approach to map the brain's connections quickly, accurately, and cost-effectively. Past efforts to address the challenge of teasing apart the complex connectome of the mammalian brain were subject to a steep trade-off between throughput/efficiency and resolution. Two cutting-edge neuronal mapping techniques—barcoding based connection mapping (BARseq) and expansion microscopy (ExM)—have proven they can achieve efficient and high-resolution connection mapping within mammalian neural tissue. We will optimize and then integrate these two techniques to map both local and long-range circuitry with a single-synapse resolution. In ExM, neural tissue is physically expanded, making it easier to disambiguate neural fibers in close proximity and to detect the precise location of synapse-associated proteins. This approach is ideally suited to teasing apart the paths and connections among densely packed local circuits. Using BARseq, neurons express unique barcoded tags, which allows even distant processes to be accurately traced to their somatic origins. Combining BARseq with in situ immunolabeling techniques, we can also precisely identify the location of synapses on each fiber. Here we propose to optimize the combination of these two approaches, which will enable a platform for generating a brainwide microconnectome with single-synapse level resolution. Success in this effort has clear implications for the future of neuroscience research, including the potential to transform our understanding of both normal brain circuitry and the specific disruptions that occur within the context of neuropsychiatric disorders.

项目摘要/摘要 该提案的总体目标是开发一种强大的方法来绘制大脑的联系 快速，准确和成本效益。过去的努力应对挑战综合大楼的挑战 哺乳动物大脑的连接组在吞吐量/效率和 解决。两种尖端的神经元映射技术 - 基于Barcoding的连接映射（BARSEQ） 和扩展显微镜（EXM） - 经过证明，它们可以实现高效且高分辨率的连接 在哺乳动物神经组织中映射。我们将优化，然后集成这两种技术以映射 局部和远程电路，具有单共溶液的分辨率。在EXM中，神经组织在物理上是 扩展，使其更容易在近距离接近神经纤维并检测到确切位置的歧义 突触相关的蛋白质。这种方法非常适合嘲笑分开的路径和联系 使用barseq，神经元表达独特的条形码标签，它允许 遥远的过程可以准确地追溯到其躯体起源。将barseq与原位结合 免疫标记技术，我们还可以准确地识别每种纤维上突触的位置。我们在这里 提议优化这两种方法的组合，这将使一个平台生成一个 带有单共同点水平分辨率的Brahewide Microconneconeconme。这项工作的成功具有明显的影响 对于神经科学研究的未来，包括改变我们对两者正常的理解的潜力 脑电路和在神经精神疾病背景下发生的特定破坏。