Scalable integration of cell types and connectivity in the motor cortex of rodents and non-human primates

啮齿类动物和非人类灵长类动物运动皮层中细胞类型和连接性的可扩展整合

• 批准号: 10369307
• 负责人: Forrest Christie Collman
• 金额: $ 415.4万
• 依托单位: ALLEN INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369307
• 关键词: 3-Dimensional; Address; Adoption; Anatomy; Area; Axon; BRAIN initiative; Brain; Cell Therapy; Cells; Cellular Morphology; Cellular Structures; Censuses; Classification; Data; Data Set; Databases; Detection; Electron Microscopy; Electrons; Electrophysiology (science); Gene Delivery; Gene Expression; Genes; Genetic; Gold; Human; Image; Image Analysis; Imagery; Individual; Institutes; Label; Language; Link; Macaca; Maps; Measures; Methods; Microscopic; Modality; Modeling; Molecular; Morphology; Motor Cortex; Movement; Mus; Muscle; Neurons; Pathway interactions; Pattern; Primates; Resolution; Resources; Rodent; Role; Sampling; Source; Structure; Surveys; Synapses; Testing; Tracer; Transgenic Organisms; Viral; Virus; Work; base; cell type; connectome; cost; design; experience; experimental study; genetic technology; image processing; imaging approach; improved; in vivo; multimodality; nano; neural circuit; nonhuman primate; patch sequencing; promoter; prospective; reconstruction; tool; transcriptomics; virus genetics

Project Summary The BICCN has recently completed a broad survey of the cellular components of motor cortex, including transcriptomic profiling, patch-seq, multiplexed FISH, inter-areal connectivity, and single neuron morphology. Missing from this view is a detailed picture of how individual neurons and neuronal types interconnect, in large part because acquiring a comprehensive picture of individual neuronal connections is best achieved with the difficult methods of large-scale electron microscopic reconstructions. Further, there are no scalable methods to assign transcriptomic cell classes to EM reconstructions, particularly when considering the myriad set of non- local inputs. In order to fulfill the BICCN mandate, we need scalable methods for measuring connectivity that can be integrated with cross-modal data, such as from Patch-seq, or with tools that target specific cell classes in transgenic and non-transgenic species, such as non-human primate. Here, we propose to exploit both approaches—retrospectively linking EM data with Patch-seq datasets or the prospective targeting of cell classes labeling in EM reconstructions—with our large-scale electron microscopy pipeline, while continuing to improve throughput and lower its overall cost. We propose to deploy these tools to examine the motor cortex of mouse and non-human primate, to demonstrate a scalable approach to delivering data that integrate local cellular morphology, ultrastructural detail, and specific local connectivity with transcriptomic information, from Patch-seq, while identifying the source and cell type of individual afferents, with viral genetic tools.

项目摘要 BICCN最近完成了对运动皮层细胞成分的广泛调查，包括 转录组学分析、patch-seq、多重FISH、区域间连接和单神经元形态。 这种观点缺少的是单个神经元和神经元类型如何相互连接的详细图片， 部分原因是，获得单个神经元连接的全面图像最好是通过 大规模电子显微镜重建的困难方法。此外，没有可扩展的方法来 将转录组细胞分类分配给EM重建，特别是当考虑到无数的非转录组细胞时， 当地投入。为了完成BICCN的任务，我们需要可扩展的方法来测量连接性， 可以与跨模态数据（如Patch-seq）集成，也可以与针对特定细胞类的工具集成 在转基因和非转基因物种如非人灵长类中。在这里，我们建议利用这两个 方法-回顾性地将EM数据与Patch-seq数据集或细胞类别的前瞻性靶向联系起来 EM重建中的标记-使用我们的大规模电子显微镜管道，同时继续改进 生产能力并降低其总成本。我们建议使用这些工具来检查小鼠的运动皮层 和非人类灵长类动物，以展示一种可扩展的方法来提供数据， 形态学、超微结构细节和与转录组学信息的特定局部连接，来自Patch-seq， 同时用病毒遗传工具识别个体传入的来源和细胞类型。