Molecular Science Core

分子科学核心

• 批准号: 10294400
• 负责人: Bosiljka Tasic
• 金额: $ 44.05万
• 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/10294400
• 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 Analyses; Data Set; Databases; Dependovirus; Diffuse; Enhancers; Fluorescent in Situ Hybridization; Gene Expression; Genes; Genetic; Genetic Enhancer Element; Genomics; Goals; Hippocampus (Brain); Institutes; Knock-in; Label; Logic; 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; base; bioinformatics tool; candidate marker; cell type; cellular imaging; epigenomics; excitatory neuron; genetic approach; genetic element; genomic data; improved; molecular marker; 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，接收来自丘脑以外不同区域的输入 （新皮质、小脑、基底神经节、中脑、海马体）并投射到新皮质。最近的分子 绘图和形态学重建表明，这些丘脑皮质 (TC) 细胞比 在细胞结构定义的丘脑核之间和内部，比之前想象的要多样化。不同的 TC 细胞可能接收来自不同皮质下结构的输入，并投射到不同的皮质区域和层。 实验获取特定 TC 细胞类型对于阐明它们在神经回路功能中的作用至关重要。 单细胞基因组学最近在分子水平上实现了前所未有的细胞类型定义，并已 发现了许多标记基因和推定的调控元件，可用于产生遗传 用于获取特定细胞群的工具。 对于成年小鼠丘脑，我们生成了大规模单细胞转录组数据集（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 以及用于遗传获取 TC 类型的病毒工具。为了完善对特定 TC 类型的访问，交叉遗传 将探索结合病毒、转基因和逆行标记的方法。我们将评估 标记特定细胞群的完整性与全脑水平的特异性，因为不同 不同的实验目的需要特异性/完整性水平。稀疏但具体的标签 是形态学检查的理想选择，而相对完整且局部或全局特定的标记是 行为扰动所需的。 这些工具一旦可用，将立即用于其他研究领域（项目 2、3、4）。工具 表征将结合标准化的艾伦研究所流程：全脑细胞成像、多重分析 荧光原位杂交 (mFISH) 以及 scRNA-seq。 DNA构建体、转基因小鼠、病毒 将制作包装技术、推荐的病毒滴度以及病毒和转基因特征数据 可在公共存储库中找到。