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：
9145351
负责人：
JOHN J. NGAI
金额：
$ 21.52万
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 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9145351
关键词：
Anatomy Atlases BRAIN initiative Base of the Brain Behavior Biological Biological Models Biology Brain Brain Mapping Brain region Cataloging Catalogs Cell Separation Cell physiology Cells Cellular Morphology Censuses Classification Classification Scheme Clustered Regularly Interspaced Short Palindromic Repeats 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 transcriptomics

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（由申请人提供）：揭示哺乳动物大脑的复杂性是当今生物学中最具挑战性的问题之一，神经科学的一个主要目标是了解神经元和非神经元细胞的回路如何处理感觉信息、产生运动和产生运动。阐明神经回路的特性需要了解构成这些回路的细胞类型及其在处理和整合信息中的作用。然而，自从一个多世纪以来对不同神经细胞类型的描述。在拉蒙·卡哈尔（Ramon y Cajal）之前，我们对大脑中细胞类型的多样性以及每种细胞类型如何对神经系统功能做出贡献仅触及了表面。目前对神经元进行分类的方法依赖于包括少量神经元差异表达在内的特征。基因、细胞形态、解剖位置、生理学和连接性——重要的描述性属性，但不足以完全描述或预测构成哺乳动物大脑的大量不同细胞类型。在这里，我们提出了一套用于识别和分类的技术。存在于体内的多种细胞类型我们的方法将使用来自小鼠体感皮层的第 5 层锥体细胞作为模型系统来开发，我们将利用 DNA 测序技术的最新发展来表征单层 5 神经元的基因表达谱。其次，定义新发现的神经元亚型的基因将用于通过 Cas9/CRISPR 介导的基因组工程获得这些细胞的遗传途径，以创建转基因报告系。这项技术有望为快速生成多基因小鼠报告品系开辟一条途径，其中特定的神经元亚型由标记基因的组合进行独特标记。第三，我们将使用这些基因工程小鼠来确认我们的分类法代表了不同的功能特性。我们的方法最终可以扩大规模，以生成大脑中细胞类型的完整普查，这是利用现代研究工具剖析神经系统功能所急需的资源。