The landscape of mouse epididymal cells defined by the single-cell RNA-Seq. The landscape of mouse epididymal cells defined by the single-cell RNA-Seq

We reported the cell composition and segmental expression differences in three epididymis segments using single-cell RNA sequencing. A total of 13,949 cells were detected, which were further clustered into eight cell populations identified with all known constituent cells of mouse epididymis including principle cell, basal cell, clear/narrow cell and immune cells. Through the study of 35 known epididymal proteins linking to sperm function phenotypes (EPSFPs), we found they were mainly expressed in principle cells and also revealed their segmental specific expression patterns. Notably, a part of EPSFPs was associated with sperm protection and predominantly localized in caput principal cells, whereas some EPSFPs, highly expressed in corpus and cauda, were served as the regulating molecular for the sperm functions such as motility, capacitation or acrosome reactions.In addition, focused analyses uncovered 5 subtypes of the principal cell. Intriguingly, Prc5, the subpopulation of principle cells, was almost only existed in the caput epididymis. Overall design: 56 days old of wild-type C57Bl/6J mice were sacrified, then the epididymis were dissected and divided into three regions (caput, corpus, and cauda)

本研究通过单细胞RNA测序（single-cell RNA sequencing），解析了小鼠附睾三个节段的细胞组成及节段间表达差异。本次研究共检测到13949个细胞，经聚类后分为8个细胞群，涵盖了小鼠附睾已报道的所有固有细胞类型，包括主细胞（principal cell）、基底细胞（basal cell）、亮/窄细胞及免疫细胞。通过对35种与精子功能表型相关的已知附睾蛋白（epididymal proteins linking to sperm function phenotypes，EPSFPs）进行分析，本研究发现此类蛋白主要表达于主细胞中，并揭示了它们的节段特异性表达模式。值得注意的是，部分EPSFPs与精子保护相关，且主要定位于附睾头部（caput）主细胞；而另一些在附睾体部（corpus）和尾部（cauda）高表达的EPSFPs，则可作为调控分子参与精子运动、获能及顶体反应等功能过程。此外，针对性分析还揭示了主细胞的5个亚型。有趣的是，主细胞亚群Prc5几乎仅存在于附睾头部。实验设计：选取56日龄的野生型C57Bl/6J小鼠实施安乐死，随后解剖其附睾并分为三个节段（附睾头、附睾体及附睾尾）。