A branching model of lineage differentiation underpinning the neurogenic potential of enteric glia (Single-cell ATAC-seq data for enteric glial cells). A branching model of lineage differentiation underpinning the neurogenic potential of enteric glia (Single-cell ATAC-seq data for enteric glial cells)

Glial cells have been proposed as an endogenous source of progenitors for the treatment of neural deficits. However, the cellular and molecular mechanisms underpinning the neurogenic potential of certain populations of adult glial cells, are not known. Using single cell transcriptomic profiling, we show here that enteric glial cells represent a cell state attained by autonomic neural crest cells as they transition during development along a linear default differentiation trajectory that allows them to retain neurogenic potential while acquiring a gene expression profile associated with their role in neuronal support and immunomodulation. Key neurogenic loci in early enteric nervous system progenitors remain in open chromatin configuration in mature enteric glia, thus facilitating neuronal differentiation under appropriate conditions. Molecular profiling and gene targeting of enteric glial cells in a novel cell culture system of enteric neurogenesis and a gut injury model, demonstrated that neuronal differentiation of glia is driven by transcriptional programs employed in vivo by early progenitors. Our work provides mechanistic insight into the dynamic regulatory landscape underpinning the development of intestinal neural circuits and generates a platform for advancing glial cells as therapeutic agents for the treatment of neural deficits. Overall design: We performed scATAC-seq of nGFP+ cells isolated from tunica muscularis preparations from 8 week old Sox10CreERT2(SER93);Rosa26-nuclearGFP mice that had been injected with tamoxifen (2 doses of tamoxifen at 100 μg/g of body weight 10 days prior isolation).

胶质细胞（glial cells）已被提议作为治疗神经功能缺损的内源性祖细胞来源。然而，特定亚群成年胶质细胞的神经发生潜能所依赖的细胞与分子机制，目前仍未阐明。
本研究借助单细胞转录组测序（single cell transcriptomic profiling）技术，证实肠胶质细胞（enteric glial cells）是自主神经嵴细胞（autonomic neural crest cells）在发育过程中沿线性默认分化轨迹转变所获得的细胞状态；该轨迹使其既能保留神经发生潜能，又能获得与神经元支持及免疫调节功能相关的基因表达谱。早期肠神经系统祖细胞中的关键神经发生基因座，在成熟肠胶质细胞中仍保持开放染色质构象，因此在适宜条件下可促进神经元分化。
本研究通过肠神经发生新型细胞培养系统与肠道损伤模型，对肠胶质细胞开展分子谱分析与基因靶向实验，证实胶质细胞的神经元分化过程由早期祖细胞体内激活的转录程序所驱动。本研究为阐明肠道神经环路发育的动态调控机制提供了机制层面的见解，并构建了以胶质细胞作为治疗神经功能缺损的治疗剂的研究平台。
整体实验设计：我们对分离自8周龄Sox10CreERT2(SER93);Rosa26-核GFP小鼠肌层制剂的nGFP+细胞进行了单细胞转座酶可及性测序（scATAC-seq）；这些小鼠在分离细胞前10天接受他莫昔芬注射（剂量为每克体重100 μg他莫昔芬，共2次）。