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.

项目概要/摘要 该提案的首要目标是开发一种强大的方法来绘制大脑的连接图 快速、准确且经济高效地应对梳理复杂结构的挑战。 哺乳动物大脑的连接组受到吞吐量/效率和 两种尖端的神经映射技术——基于条形码的连接映射（BARseq） 和扩展显微镜（ExM）——已证明它们可以实现高效、高分辨率的连接 我们将优化并整合这两种技术来绘制哺乳动物神经组织内的图谱。 具有单突触分辨率的局部和远程电路在 ExM 中，神经组织是物理上的。 扩展，使得更容易消除邻近神经纤维的歧义并检测神经纤维的精确位置 这种方法非常适合梳理突触相关蛋白之间的路径和连接。 使用密集的局部电路 BARseq，神经元可以表达独特的条形码标签，甚至可以实现。 将 BARseq 与原位相结合，可以准确追踪遥远的过程。 通过免疫标记技术，我们还可以精确识别每根纤维上突触的位置。 建议优化这两种方法的组合，这将使一个平台能够生成 具有单突触水平分辨率的全脑微连接组这项工作的成功具有明确的意义。 神经科学研究的未来，包括改变我们对正常神经科学研究的理解的潜力 大脑回路和神经精神疾病背景下发生的特定破坏。