Cell atlas of mouse brain-spinal cord connectome

小鼠脑脊髓连接组细胞图谱

基本信息

批准号：
9583948
负责人：
Hong-Wei Dong
金额：
$ 269.17万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9583948
关键词：
Anatomy Atlases BRAIN initiative Bar Codes Base of the Brain Biological Assay Brain Brain region Candidate Disease Gene Catalogs Cell Separation Cells Censuses Chemicals Communities Computer software Data Databases Electrophysiology (science)Gene Expression Gene Expression Profile Genes Goals Image Image Analysis In Situ Informatics Knowledge Label Laboratories Light Maps Methods Microscopy Modality Molecular Molecular Genetics Molecular Profiling Morphology Mus Names Neurons Neurosciences Online Systems Output Pathway interactions Pattern Preparation Process Property Research Resources Scanning Slice Source Spinal Spinal Cord Structure Techniques Technology Three-Dimensional Imaging Tracer Viral base brain cell candidate marker cell type collaboratory combinatorial connectome data portal experimental study in vivo innovation interdisciplinary approach molecular marker online resource open data open source optogenetics patch clamp response single cell sequencing spinal cord imaging transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY Although great efforts have been dedicated to characterizing neuronal cell types in the brain, systematic studies on the brain-spinal cord connectome and associated spinal neuronal types are lacking. In this project, a team of seven laboratories proposes to use a highly innovative and multidisciplinary approach to systematically characterize neuronal types in the spinal cord based on their anatomy, connectivity, neuronal morphologies, molecular identities, and electrophysiological properties. In Aim 1, we will use a newly developed AAV anterograde transsynaptic tagging method to label spinal cord neurons that receive descending inputs from different brain regions, and use a retrograde viral tracer, AAVretro, to label spinal neurons that project to defined brain regions. These tagged neurons will be imaged in the intact whole spinal cord with a newly developed fast 3D light sheet microscopy technique, and targeted for recording in slice preparations. The axonal collateral patterns, dendritic morphologies, and electrophysiological properties will be compared between different input/output-defined spinal neuron groups. In Aim 2, the gene expression patterns of the tagged neurons will be determined in situ by sequential bar-coded FISH (seqFISH), with candidate marker genes obtained from online resources, or from single-cell sorting and RNA sequencing (Dropseq). In Aim 3, all collected data on connectivity, anatomical cell type distribution map, neuronal morphologies, molecular identities, and electrophysiological properties will be used for classifying spinal neuron types connected with brain, and an open-source data portal will be established which will allow users to search, view, and analyze the multi-modal and integrative cell-type specific data. Together, we aim to construct a comprehensive cell- type atlas of the mouse brain-spinal cord connectome.

项目概要尽管人们付出了巨大的努力来表征大脑中的神经元细胞类型，但系统性的缺乏对脑脊髓连接组和相关脊髓神经元类型的研究。在这个项目中，一个由七个实验室组成的团队建议使用高度创新和多学科的方法来系统地根据脊髓的解剖结构、连接性、神经元形态来表征脊髓中的神经元类型，分子特性和电生理特性。在目标 1 中，我们将使用新开发的 AAV 顺行突触标记方法来标记接收下行输入的脊髓神经元不同的大脑区域，并使用逆行病毒示踪剂 AAVretro 来标记投射到明确的大脑区域。这些标记的神经元将在完整的整个脊髓中使用新的成像技术进行成像。开发了快速 3D 光片显微镜技术，并针对切片制备进行记录。这将比较轴突侧支模式、树突形态和电生理特性不同输入/输出定义的脊髓神经元组之间。在目标 2 中，基因表达模式标记的神经元将通过连续条形码 FISH (seqFISH) 进行原位测定，并带有候选标记从在线资源或单细胞分选和 RNA 测序 (Dropseq) 获得的基因。在目标 3 中，所有收集有关连通性、解剖细胞类型分布图、神经元形态、分子身份和电生理特性将用于对与相关的脊髓神经元类型进行分类将建立一个开源数据门户，允许用户搜索、查看和分析多模式和综合细胞类型特定数据。我们的目标是共同构建一个全面的细胞- 小鼠脑脊髓连接组的类型图谱。