Molecular Science Core

分子科学核心

• 批准号: 10546511
• 负责人: Bosiljka Tasic
• 金额: $ 50.61万
• 依托单位: ALLEN INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-15 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10546511
• 关键词: ATAC-seq; Adult; Amygdaloid structure; Anatomy; Animals; Area; Automobile Driving; Award; Axon; BRAIN initiative; Basal Ganglia; Behavioral; Brain; Brain imaging; Cell Nucleus; Cells; Censuses; Cerebellum; Chromatin; Corpus striatum structure; DNA; Data; Data Set; Databases; Dependovirus; Diffuse; Enhancers; Fluorescent in Situ Hybridization; Gene Expression; Genes; Genetic; Genetic Enhancer Element; Genomics; Goals; Hippocampus; Knock-in; Label; Logic; Maps; Measures; Midbrain structure; Modernization; Molecular; Morphology; Mus; Neocortex; Neurons; Organ; Pattern; Population; Process; Publishing; Reagent; Recommendation; Regulatory Element; Reporter; Research; Role; Science; Specificity; Standardization; Structure; Taxonomy; Techniques; Thalamic Nuclei; Thalamic structure; Transgenes; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Viral; Viral Genome; Viral Packaging; Viral Vector; Virus; Work; bioinformatics tool; candidate marker; cell type; cellular imaging; epigenomics; excitatory neuron; genetic approach; genetic element; genomic data; improved; molecular marker; neocortical; neural circuit; public repository; recombinase; reconstruction; single-cell RNA sequencing; tomography; tool; transcription factor; transcriptomics; two-photon

Summary, Molecular Science Core The thalamus contains excitatory neurons that receive input from diverse areas outside of the thalamus (neocortex, cerebellum, basal ganglia, midbrain, hippocampus) and project to the neocortex. Recent molecular mapping and morphological reconstructions have shown that these thalamocortical (TC) cells are much more diverse than previously thought, both across and within cytoarchitectonically-defined thalamic nuclei. Different TC cells likely receive input from different subcortical structures, and project to different cortical areas and layers. Experimental access to specific TC cell types is essential for elucidating their roles in neural circuit function. Single cell genomics has recently enabled unprecedented definition of cell types at the molecular level and has uncovered numerous marker genes and putative regulatory elements, which can be used to generate genetic tools for access to specific cell populations. For the adult mouse thalamus, we have generated a large-scale single-cell transcriptomics dataset (scRNA-seq) to measure gene expression, and a matching single-nucleus chromatin accessibility dataset (snATAC-seq) to discover putative enhancers. In addition, we are in the process of generating equivalent data for the developing mouse thalamus. These datasets will enable us to select marker genes and enhancers to generate transgenic and viral tools for genetic access to TC types. To refine access to specific TC types, intersectional genetic approaches combining viruses, transgenes, and retrograde labeling will be explored. We will evaluate the completeness of labeling a specific cell population versus specificity at the level of the whole brain, as different levels of specificity / completeness are needed for different experimental purposes. Sparse yet specific labeling is ideal for morphological examination, whereas relatively complete and locally or globally specific labeling is needed for behavioral perturbations. The tools will be used in other research segments (Projects 2, 3, 4) as soon as they become available. Tool characterization will combine standardized Allen Institute pipelines: whole-brain cellular imaging, multiplexed fluorescence in situ hybridization (mFISH), as well as scRNA-seq. DNA constructs, transgenic mice, virus packaging techniques, recommended virus titers, and virus and transgene characterization data will be made available in public repositories.

摘要，分子科学核心 丘脑包含兴奋性神经元，它们接收来自丘脑外不同区域的输入 （新皮层、小脑、基底神经节、中脑、海马）并投射到新皮层。最近的分子 映射和形态重建显示，这些丘脑皮质（TC）细胞比其他细胞多得多， 多样性比以前认为的，无论是跨和cytoarchitectonically定义丘脑核内。不同 TC细胞可能从不同的皮层下结构接收输入，并投射到不同的皮层区域和层。 实验获得特定的TC细胞类型是阐明它们在神经回路功能中的作用所必需的。 单细胞基因组学最近使得能够在分子水平上前所未有地定义细胞类型，并且 发现了许多标记基因和推定的调控元件，可用于产生遗传 获取特定细胞群的工具。 对于成年小鼠丘脑，我们已经生成了一个大规模的单细胞转录组学数据集（scRNA-seq）。 测量基因表达，以及匹配的单核染色质可及性数据集（snATAC-seq）， 发现假定的增强子。此外，我们正在为开发人员生成等效数据 小鼠丘脑。这些数据集将使我们能够选择标记基因和增强子来产生转基因 和病毒工具来获取TC类型的基因。为了改善对特定TC类型的访问，交叉遗传学 将探索结合病毒、转基因和逆行标记的方法。我们将评估 标记特定细胞群的完整性与在全脑水平上的特异性， 不同的实验目的需要特异性/完整性水平。稀疏但特定的标记 是形态学检查的理想选择，而相对完整和局部或全局特异性标记， 行为紊乱所需要的。 这些工具一旦获得，将用于其他研究部分（项目2、3、4）。工具 表征将结合联合收割机标准化的艾伦研究所管道：全脑细胞成像，多路复用 荧光原位杂交（mFISH），以及scRNA-seq. DNA构建体，转基因小鼠，病毒 包装技术，推荐的病毒滴度，以及病毒和转基因的特征数据 在公共仓库中可用。