Transcriptomes of major renal collecting duct cell types in mouse identified by single-cell RNA-seq
Transcriptomes of major renal collecting duct cell types in mouse identified by single-cell RNA-seq
复制标题
DOI：
10.1073/pnas.1710964114
复制
发表时间：
2017-11-14
影响因子：
11.1
通讯作者：
Knepper, Mark A.
中科院分区：
文献类型：
--
作者：
Chen, Lihe;Lee, Jae Wook;Knepper, Mark A.
先前的RNA测序(RNA-seq)研究已经确定了大多数肾上皮细胞类型的完整转录本。例外的是组成肾脏集合管的细胞类型，即间质细胞(ICs)和主细胞(PC)，它们只占肾脏质量的一小部分，但在调节血压、细胞外液容量和细胞外液成分方面发挥着关键的生理作用。为了丰富这些细胞类型，我们使用了FAC，它使用了针对PC和B型IC的成熟的凝集素细胞表面标记，以及新发现的用于A型IC的细胞表面标记c-Kit。使用IC和PC富集群作为输入的单细胞RNA-SEQ能够识别A-IC、B-IC和PC的完整转录本。这些数据被用来创建一个免费访问的在线基因表达数据库，用于收集管道细胞。这个数据库可以识别在每种细胞类型中选择性表达的基因，包括细胞表面受体、转录因子、转运体和分泌蛋白。分析还鉴定了一小部分杂交细胞表达水通道蛋白-2和阴离子交换器1或垂蛋白转录本。在许多情况下，受体及其配体的mRNAs在不同的细胞中被发现(例如，Notch2主要在PC中，Jag1主要在ICs中)，这表明这三种细胞类型之间存在信号串扰。三种集合管细胞基因表达谱的确定为了解集合管的生理调节和病理生理学奠定了基础。
Prior RNA sequencing (RNA-seq) studies have identified complete transcriptomes for most renal epithelial cell types. The exceptions are the cell types that make up the renal collecting duct, namely intercalated cells (ICs) and principal cells (PCs), which account for only a small fraction of the kidney mass, but play critical physiological roles in the regulation of blood pressure, extracellular fluid volume, and extracellular fluid composition. To enrich these cell types, we used FACS that employed well-established lectin cell surface markers for PCs and type B ICs, as well as a newly identified cell surfacemarker for type A ICs, c-Kit. Single-cell RNA-seq using the IC-and PC-enriched populations as input enabled identification of complete transcriptomes of A-ICs, B-ICs, and PCs. The data were used to create a freely accessible online gene-expression database for collecting duct cells. This database allowed identification of genes that are selectively expressed in each cell type, including cell-surface receptors, transcription factors, transporters, and secreted proteins. The analysis also identified a small fraction of hybrid cells expressing aquaporin-2 and anion exchanger 1 or pendrin transcripts. In many cases, mRNAs for receptors and their ligands were identified in different cells (e.g., Notch2 chiefly in PCs vs. Jag1 chiefly in ICs), suggesting signaling cross-talk among the three cell types. The identified patterns of gene expression among the three types of collecting duct cells provide a foundation for understanding physiological regulation and pathophysiology in the renal collecting duct.