Primary antibodies for immunohistochemistry.

Peripheral glia are important regulators of diverse physiologic functions yet their molecular distinctions and locations in almost all visceral organs are not well-understood. We performed a systematic analysis of peripheral glia, focusing on the lung and leveraging single cell RNA sequencing (scRNA-seq) analysis to characterize their cellular and molecular features. Using in vivo lineage studies, we characterized the anatomic, cellular, and molecular features of the Sox10+ glial lineage of the mouse lung. Using high-resolution imaging, we quantified the distribution and cellular morphologies of myelinating, non-myelinating, satellite, and terminal glial cells with their intricate extensions along peripheral nerves, including terminals at specialized neurosensory structures within the lung. Spatial analysis of selectively expressed myelinating (periaxin/Prx, claudin 19/Cldn) or non-myelinating (sodium channel/Scn7a) glial cell genes identified by scRNA-seq analysis revealed molecularly distinct populations surrounding myelinated nerve fibers in the lung. To extend this analysis to primates and other organs, we extracted rare peripheral glial cells in whole organism scRNA-seq atlases of mouse lemur and human. Our cross-species data analysis and integration of scRNA-seq data of ~700 peripheral glial cells from mouse, mouse lemur, and human glial cells identified conserved gene expression of molecularly distinct peripheral glial cell populations. This foundational knowledge facilitates subsequent functional studies targeting molecularly distinct subsets of peripheral glia and integrating them into organ-specific disorders of autonomic dysregulation. In addition, our cross-species analysis identifying conserved gene expression patterns and glial networks in extrapulmonary organs provides a valuable resource for studying the functional role of peripheral glia in multiorgan human diseases.

外周胶质细胞是多种生理功能的重要调控因子，但目前学界对几乎所有内脏器官中外周胶质细胞的分子特征差异及其分布位置仍缺乏充分了解。本研究聚焦肺组织，对其中的外周胶质细胞开展系统性分析，借助单细胞RNA测序（single cell RNA sequencing, scRNA-seq）技术解析其细胞与分子特征。通过体内谱系示踪实验，我们解析了小鼠肺组织中Sox10阳性胶质细胞谱系的解剖学、细胞及分子特征。借助高分辨率成像技术，我们定量分析了有髓鞘胶质细胞、无髓鞘胶质细胞、卫星胶质细胞及终末胶质细胞的分布特征与细胞形态，及其沿外周神经延伸的复杂突起结构，包括肺内特定神经感觉结构处的神经末梢。对单细胞RNA测序筛选出的髓鞘相关胶质细胞特异性表达基因（periaxin/Prx、claudin 19/Cldn）与无髓鞘相关胶质细胞特异性表达基因（钠通道/Scn7a）开展空间分析，结果显示肺内有髓神经纤维周围存在分子特征迥异的胶质细胞亚群。为将该分析拓展至灵长类动物及其他器官，我们从小鼠狐猴与人类的全机体单细胞RNA测序图谱中分离得到稀有外周胶质细胞。本研究对小鼠、小鼠狐猴及人类来源的约700个外周胶质细胞的单细胞RNA测序数据进行跨物种数据分析与整合，鉴定出分子特征各异的外周胶质细胞亚群的保守基因表达谱。这一基础性研究成果可为后续针对特定分子亚群的外周胶质细胞功能研究提供支撑，并助力将其应用于自主神经失调相关器官特异性疾病的研究。此外，本研究通过跨物种分析鉴定出肺外器官中保守的基因表达模式与胶质细胞调控网络，为探究外周胶质细胞在多器官人类疾病中的功能作用提供了宝贵的研究资源。