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最近对运动皮层的细胞成分进行了广泛的调查，包括 转录组分析，斑块序列，多路复用鱼类，美园连通性和单个神经元形态。 从这种角度看，缺少的是单个神经元和神经元类型如何互连的详细图片 部分是因为最好通过获得单个神经元联系的全面图片。 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 将转录组细胞类分配给EM重建，尤其是在考虑无数的非 - 本地输入。为了履行BICCN任务，我们需要可扩展的方法来衡量连接性 可以与跨模式数据集成，例如来自patch-seq，也可以与针对特定单元格类的工具集成 在转基因和非转基因物种中，例如非人类灵长类动物。在这里，我们建议利用这两个 方法 - 将EM数据与补丁程序数据集或单元类的前瞻性靶向链接性链接 在EM重建中标记 - 我们的大型电子显微镜管道，同时继续改进 吞吐量并降低其整体成本。我们建议部署这些工具来检查鼠标的运动皮层 和非人类灵长类 形态学，超微结构细节和特定的局部连通性，带有转录组信息，patch-seq， 同时使用病毒遗传工具来识别个体传入的源和细胞类型。