Molecular Science Core

分子科学核心综述 丘脑含有兴奋性神经元，从丘脑以外的不同区域接受输入。 (新皮质、小脑、基底节、中脑、海马体)并投射到新皮质。最新分子 作图和形态重建表明，这些丘脑皮质(TC)细胞 与之前认为的不同，无论是在细胞结构定义的丘脑核团之间还是在细胞结构定义的丘脑核团内。不同 TC细胞可能接受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。 通过实验获得特定类型的TC细胞对于阐明它们在神经回路功能中的作用是至关重要的。 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 发现了大量的标记基因和假定的调控元件，可以用来产生基因 获取特定细胞群体的工具。 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数据集(scrna-seq)。 以测量基因表达，以及匹配的单核染色质可及性数据集(SnATAC-seq)以 寻找可能的增强剂。此外，我们正在生成用于开发的等效数据 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。要细化对特定TC类型的访问，交叉遗传 将探索结合病毒、转基因和逆行标记的方法。我们将评估 标记特定细胞群体的完整性与整个大脑水平的特异性不同 不同的实验目的需要不同程度的特异性/完整性。稀疏但具体的标注 是形态检查的理想选择，而相对完整和局部或全局特定的标记是 行为紊乱所需要的。 这些工具一旦可用，将立即用于其他研究部门(项目2、3、4)。工具 特征化将结合标准化的艾伦研究所管道：全脑细胞成像，多路传输 荧光原位杂交(MFISH)和scRNA-seq.DNA构建、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。 在公共存储库中可用。

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.