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

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.

Emergent dynamics of adult stem cell lineages from single nucleus and single cell RNA-Seq of Drosophila testes

“…Production of functional sperm requires intimate interactions between germ cells and somatic support cells, with defects at almost any step compromising fertility. Interest in spermatogenesis has motivated scRNA-seq analyses of testes from a variety of organisms, including mouse ( Cao et al, 2021 ; Chen et al, 2018 ; Green et al, 2018 ; Guo et al, 2020 ; Law et al, 2019 ) and Drosophila ( Li et al, 2022 ; Mahadevaraju et al, 2021 ; Witt et al, 2019 ). Notably, the testis of Drosophila has the highest complexity in terms of mRNAs expressed of any tissue in the fly, likely reflecting the dramatic differentiation events required ( Li et al, 2022 ).…”

Knockout of cyclin dependent kinases 8 and 19 leads to depletion of cyclin C and suppresses spermatogenesis and male fertility in mice

“…Sertoli cells are the main driver of spermatogenic differentiation that receive testosterone The SCARKO mice were engineered two decades ago and show morphological abnormalities (i.e, reduced testes, epididymis and prostate, meiotic arrest during meiosis I prophase) similar to DKO [59,60]. scRNAseq technique used to elucidate the molecular mechanism of the meiotic arrest revealed a number of similarities with DKO single-cell RNA seq data for Sertoli cells, especially in terms of pathways [61]. Thus, genes associated with cytoskeleton, cellular membrane and cell-cell contacts were enriched among the DEGs (Fig.…”

“…For stable cluster assignment, a reference dataset was formed of WT samples, which were integrated via Harmony (Korsunsky et al, 2018) and then clustered with Seurat. Clustering was checked against testis cell type markers (Cao et al, 2021; Hermann et al, 2018) to ensure selection of true clusters that would reiterate across samples. Cluster of cells belonging to the IGB_2 WT sample only, which did not correspond to any cell type and was missing essential housekeeping genes, was discarded as a cluster of low quality cells with no biological relevance.…”

“…Visualizations were also made with the Seurat package. Cell type was assigned according to their marker genes (Cao et al, 2021; Hermann et al, 2018). Both cells and microenvironment in the testis were detected in wild-type as well as in KO samples.…”

Single-cell transcriptomics reveals male germ cells and Sertoli cells developmental patterns in dairy goats

“…Interestingly, scRNA-Seq technology can be used to explore cell heterogeneity and transcriptional changes that occur during spermatogenesis (Lau et al, 2020). Currently, single-cell transcriptomics of spermatogenesis has mainly focused on the testis cells of humans and mice (Wang et al, 2018;Cao et al, 2021). The molecular functions of spermatogonia, spermatocytes, spermatids, and testicular somatic cells as well as the heterogeneity among these cells have been reported (Suzuki et al, 2019).…”