Classification of Cortical Neurons by Single Cell Transcriptomics

单细胞转录组学对皮质神经元的分类

基本信息

批准号：
9107492
负责人：
JOHN J. NGAI
金额：
$ 146.13万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-26 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9107492
关键词：
Anatomy Atlases BRAIN initiative Base of the Brain Behavior Biological Biological Models Biology Brain Brain Mapping Brain region CRISPR/Cas technology Cataloging Catalogs Cell Separation Cell physiology Cells Cellular Morphology Censuses Classification Classification Scheme Cognition Complement Complex Conscious DNA Sequence Development Emotions Equipment and supply inventories Fingerprint Future Gene Expression Gene Expression Profile Generations Genes Genetic Genetic Techniques Genetically Engineered Mouse Genome Genome engineering Genotype Goals Health Human In Situ Individual Label Location Logic Maps Mediating Memory Methods Molecular Molecular Profiling Morphology Motor output Mouse Strains Movement Mus Nervous System Physiology Neuroglia Neurons Neurosciences Organ Organism Pattern Physiological Physiology Population Process Property Proteins Pyramidal Cells RNA Reporter Research Resources Role Sampling Sensory Sensory Process Somatosensory Cortex Staging Statistical Methods Surface Taxonomy Technology Testing Tissues Transgenic Mice Transgenic Organisms Validation base cell type differential expression hippocampal pyramidal neuron insight molecular marker neural circuit optogenetics recombinase research study scale up single cell sequencing technology development tool transcriptome transcriptomics

Anatomy Atlases BRAIN initiative Base of the Brain Behavior Biological Biological Models Biology Brain Brain Mapping Brain region CRISPR/Cas technology Cataloging Catalogs Cell Separation Cell physiology Cells Cellular Morphology Censuses Classification Classification Scheme Cognition Complement Complex Conscious DNA Sequence Development Emotions Equipment and supply inventories Fingerprint Future Gene Expression Gene Expression Profile Generations Genes Genetic Genetic Techniques Genetically Engineered Mouse Genome Genome engineering Genotype Goals Health Human In Situ Individual Label Location Logic Maps Mediating Memory Methods Molecular Molecular Profiling Morphology Motor output Mouse Strains Movement Mus Nervous System Physiology Neuroglia Neurons Neurosciences Organ Organism Pattern Physiological Physiology Population Process Property Proteins Pyramidal Cells RNA Reporter Research Resources Role Sampling Sensory Sensory Process Somatosensory Cortex Staging Statistical Methods Surface Taxonomy Technology Testing Tissues Transgenic Mice Transgenic Organisms Validation base cell type differential expression hippocampal pyramidal neuron insight molecular marker neural circuit optogenetics recombinase research study scale up single cell sequencing technology development tool transcriptome transcriptomics

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项目摘要

DESCRIPTION (provided by applicant):Unraveling the complexity of the mammalian brain is one of the most challenging problems in biology today. A major goal of neuroscience is to understand how circuits of neurons and non-neuronal cells process sensory information, generate movement, and subserve memory, emotion and cognition. Elucidating the properties of neural circuits requires an understanding of the cell types that comprise these circuits and their roles in processing and integrating information. However, since the description of diverse neuronal cell types over a century ago by Ramon y Cajal, we have barely scratched the surface of understanding the diversity of cell types in the brain and how each individual cell type contributes to nervous system function. Current approaches for classifying neurons rely upon features including the differential expression of small numbers of genes, cell morphology, anatomical location, physiology, and connectivity - important descriptive properties that nonetheless are insufficient to fully describe or predict the vast number of different cell types that comprise the mammalian brain. Here we propose a suite of technologies for identifying and classifying the myriad cell types present in the brain. Our method will be developed using layer 5 pyramidal cells from mouse somatosensory cortex as a model system. First, we will exploit the latest developments in DNA sequencing technologies to characterize gene expression profiles on single layer 5 neurons at high throughput. This information will be used to classify individual cells based on their transcriptome "fingerprints." Second, genes found to define newly discovered neuronal subtypes will be used to gain genetic access to these cells using Cas9/CRISPR-mediated genome engineering to create transgenic reporter lines. Development of this technology promises to open a pipeline for the rapid generation of multigenic mouse reporter strains in which specific neuronal subtypes are uniquely labeled by combinations of tagged genes. Third, we will use these genetically engineered mice to confirm that our taxonomy represents distinct functional properties of the classified neurons. Our approach can ultimately be scaled up to generate a complete census of cell types in the brain, a critically needed resource for dissecting nervous system function with modern investigative tools.

描述（应用程序提供）：揭开哺乳动物大脑的复杂性是当今生物学中最挑战的问题之一。神经科学的主要目标是了解神经元和非神经元细胞的电路如何处理感官信息，产生运动并提供记忆，情感和认知。阐明神经元的特性需要了解完成这些电路的细胞类型及其在处理和集成信息中的作用。但是，由于Ramon Y Cajal在一个世纪前对不同神经元细胞类型的描述，我们几乎没有划过了解大脑中细胞类型的多样性以及每种单个细胞类型如何有助于神经系统功能的表面。当前进行神经元分类的方法取决于特征，包括少量基因，细胞形态，解剖位置，生理和连通性的差异表达 - 尽管如此，这些重要特性仍然不足以完全描述或预测大量伴有哺乳动物大脑的不同细胞类型。在这里，我们提出了一套技术，用于识别和分类大脑中存在的无数细胞类型。我们的方法将使用小鼠体感皮质的5层锥体细胞作为模型系统开发。首先，我们将探索DNA测序技术中的最新发展，以表征高吞吐量的单层5神经元上的基因表达谱。此信息将根据单个单元格进行转录组“指纹”来分类。其次，发现定义新发现的神经元亚型的基因将使用CAS9/CRISPR介导的基因组工程来获得对这些细胞的遗传获取，以创建转基因报告基因。这项技术的开发有望为快速生成多基因小鼠报道菌株开放管道，其中特定的神经元亚型由标记基因的组合唯一地标记。第三，我们将使用这些一般的工程小鼠来确认我们的分类法代表了分类神经元的不同功能特性。我们的方法最终可以扩大规模，以产生大脑中细胞类型的完整普查，这是一种用现代研究工具剖析神经系统功能的迫切需要的资源。