Single-Cell RNA Sequencing Defines the Regulation of Spermatogenesis by Sertoli-Cell Androgen Signaling.

Congcong Cao,Shaomei Mo,Qunlong Liu,Jing Ye,Ge Shu,Yaoting Gui,Qian Ma

doi:10.3389/fcell.2021.763267

Abstract

Androgen receptor (AR) signaling is essential for maintaining spermatogenesis and male fertility. However, the molecular mechanisms by which AR acts between male germ cells and somatic cells during spermatogenesis have not begun to be revealed until recently. With the advances obtained from the use of transgenic mice lacking AR in Sertoli cells (SCARKO) and single-cell transcriptomic sequencing (scRNA-seq), the cell specific targets of AR action as well as the genes and signaling pathways that are regulated by AR are being identified. In this study, we collected scRNA-seq data from wild-type (WT) and SCARKO mice testes at p20 and identified four somatic cell populations and two male germ cell populations. Further analysis identified that the distribution of Sertoli cells was completely different and uncovered the cellular heterogeneity and transcriptional changes between WT and SCARKO Sertoli cells. In addition, several differentially expressed genes (DEGs) in SCARKO Sertoli cells, many of which have been previously implicated in cell cycle, apoptosis and male infertility, have also been identified. Together, our research explores a novel perspective on the changes in the transcription level of various cell types between WT and SCARKO mice testes, providing new insights for the investigations of the molecular and cellular processes regulated by AR signaling in Sertoli cells.

Highlights

Spermatogenesis is a precise process in mammals, including mitosis of spermatogonial, meiosis of spermatocyte and spermiogenesis
To characterize the cellular diversity of testes, we obtained the single cell suspension from four testes of P20 WT and specific AR knockout (SCARKO) mice, respectively, and performed 10X scRNA-seq technique to identify the intercellular relationships involved in spermatogenesis (Figure 1A)
The hematoxylin-eosin staining results showed that number of meiotic germ cells was significantly reduced and surviving spermatocytes failed to progress in P20 SCARKO testis compared to WT testis, which was consistent with other studies that very few spermatocytes completing the first meiotic division in SCARKO mice testis (11, 12) (Figure 1C)

Summary

Introduction

Spermatogenesis is a precise process in mammals, including mitosis of spermatogonial, meiosis of spermatocyte and spermiogenesis. Other studies have revealed that Sertoli cell-specific AR knockout (SCARKO) mice were infertile, with spermatogenesis arrest at the diplotene stage of the first meiosis. The number of Sertoli cells and spermatogonia was not affected, spermatocytes were slightly decreased, yet postmeiotic spermatids were remarkably reduced or even absent (Chang et al, 2004; De Gendt et al, 2004; Holdcraft and Braun, 2004). These results further stressed the vital role of AR signaling for both external and internal male phenotype development

Methods

Results

Conclusion