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，这些神经元从丘脑以外的不同地区接收输入 （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 映射和形态的重建表明，这些丘脑皮质（TC）细胞更多 多样性比以前想象的，无论是在细胞结构定义的丘脑核中。不同的 TC细胞可能会从不同皮层结构中接收输入，并将其投影到不同的皮质区域和层。 对特定TC细胞类型的实验访问对于阐明其在神经回路功能中的作用至关重要。 单细胞基因组学最近已实现了分子水平细胞类型的前所未有的定义，并且具有 发现了许多标记基因和推定的调节元素，可用于产生遗传 访问特定细胞种群的工具。 对于成年小鼠丘脑，我们生成了一个大型单细胞转录组学数据集（SCRNA-SEQ） 测量基因表达和匹配的单核染色质可及性数据集（SNATAC-SEQ） 发现推定的增强剂。此外，我们正在为开发生成等效数据 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生成转基因 和病毒工具用于遗传使用TC类型。为了优化对特定TC类型的访问，交叉遗传 将探索结合病毒，转基因和逆行标签的方法。我们将评估 在整个大脑水平上标记特定细胞群体与特异性的完整性，因为 为了不同的实验目的，需要特异性 /完整性水平。稀疏而特定的标签 是形态学检查的理想选择，而相对完整，本地或全球特定的标签是 行为扰动所需。 这些工具将在其他研究段（项目2、3、4）中使用。工具 表征将结合标准化的艾伦学院管道：全脑细胞成像，多重 荧光原位杂交（m鱼）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表征数据将被制定 可提供公共存储库。