Single cell profiling reveals sex, lineage and regional diversity in adult mouse kidney

To obtain new insight into the mammalian kidney, anatomy-guided, single cell RNA sequencing was performed on the adult male and female mouse kidney. Key observations were followed up through a variety of secondary studies including genetic lineage tracing. The data document novel cell diversity, unexpected origins to key cell types, and significant sex differences centered on proximal tubules segments of the nephron and principal cells of the collecting system. A searchable database integrating these data will facilitate an understanding of gene to cell relationships in the normal and diseased mammalian kidney. With regard to generating the scRNASeq dataset, single cell dissociation was optimized for micro-dissected kidney zones [cortex, outer medulla and inner medulla ] of two adult male and female C57BL6/J mice. Single cell suspensions were processed through the 10X Genomics Chromium platform with Illumina Hi-Seq sequencing of barcoded single-cell libraries. Gene/barcode matrices were merged and further processed using the Seurat R package to perform unsupervised clustering. Profiles were annotated by matching enriched gene sets with kidney cell-type specific markers. Key observations were followed up through a variety of secondary studies including genetic lineage tracing.为深入解析哺乳动物肾脏的生物学机制，本研究针对成年雌雄小鼠肾脏开展了解剖学引导下的单细胞RNA测序（single cell RNA sequencing）。针对核心观测结果，研究团队通过包括遗传谱系示踪（genetic lineage tracing）在内的多种后续辅助实验进行了验证与拓展分析。 本数据集记录了全新的细胞多样性、关键细胞类型的意外起源，以及以肾单位近端小管段与集合系统主细胞为核心的显著性别差异。 整合上述数据的可检索数据库，将有助于研究者深入理解正常与病变状态下哺乳动物肾脏内基因与细胞类型间的调控关联。 在构建单细胞RNA测序数据集时，研究人员针对两只成年雌雄C57BL6/J小鼠的显微解剖肾脏区域[皮质（cortex）、外髓质（outer medulla）与内髓质（inner medulla）]优化了单细胞解离流程。 单细胞悬液通过10X Genomics Chromium平台进行处理，并使用Illumina Hi-Seq对带条形码的单细胞文库完成测序。 合并基因/条形码矩阵后，研究团队借助Seurat R包（Seurat R package）完成了无监督聚类分析。 通过将富集基因集与肾脏细胞类型特异性标记物进行匹配，对细胞转录组图谱进行了注释。 针对核心观测结果，研究团队再次通过包括遗传谱系示踪在内的多种后续辅助实验进行了验证与拓展分析。