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Single-cell RNA-seq of NR5A1-eGFP positive cells of the developing mouse testis from E10.5 to E16.5

ABSTRACT: Purpose: We performed a time-course single-cell RNA-seq of the somatic cells of the XY mouse gonads to study the cell population heterogeneity and the genetic program during their differentiation. Methods: We collected gonads from NR5A1-eGFP transgenic embryos at five embryonic stages: E10.5, E11.5, E12.5, E13.5, E16.5. Cells were captured with the C1 autoprep system and cDNA sequenced with Illumina HiSeq 2000. Results: One cell population was detected at E10.5 and gives rise to both Sertoli and fetal Leydig cells. A precursor cell population remains undifferentiated at E16.5 and are likely to be adult Leydig cell precursors. Conclusion: Our study is, to date, the most granular transcriptomic study of the developing mouse testis and provide a more complete model of somatic cell differentiation during male sex determination.

ORGANISM(S): Mus musculus

PROVIDER: GSE97519 | GEO | 2018/02/06

REPOSITORIES: GEO

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Project description:The nuclear receptor subfamily 5 group A member 1 (NR5A1), encoding steroidogenic factor 1 (SF-1), has been identified as a critical factor in gonadal development in animal studies. A previous study of ours suggested that upregulation of NR5A1 during early gonadal differentiation in male (46,XY) human pluripotent stem cells steers the cells into a more mature gonadal cell type. However, the detailed role of NR5A1 in female gonadal differentiation has yet to be determined. In this study, by combining the processes of gonadal differentiation and conditional gene activation, we show that NR5A1 induction predominantly upregulates the female gonadal marker inhibin subunit α (INHA) and steroidogenic markers steroidogenic acute regulatory protein (STAR), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), cytochrome P450 family 17 subfamily A member 1 (CYP17A1), hydroxy-delta-5-steroid dehydrogenase (HSD3B2) and hydroxysteroid 17-beta dehydrogenase 1 (HSD17B1). In contrast, NR5A1 induction did not seem to affect the bipotential gonadal markers gata binding protein 4 (GATA4) and Wilms tumour suppressor 1 (WT1) nor the female gonadal markers r-spondin 1 (RSPO1) and wnt family member 4 (WNT4). Differentially expressed genes were highly associated with adrenal and ovarian steroidogenesis pathways. Moreover, time-series analysis revealed different dynamic changes between male and female -induced samples, where continuously upregulated genes in female gonadal differentiation were mostly associated with adrenal steroidogenesis. Thus, in contrast to male gonadal differentiation, NR5A1 is necessary but not sufficient to steer human embryonic stem cell (hESC)-derived bipotential gonadal-like cells towards a more mature somatic, female cell fate. Instead, it seems to direct bipotential gonadal-like cells more towards a steroidogenic-like cell population. The information obtained in this study helps in elucidating the role of NR5A1 in gonadal differentiation of a female stem cell line.

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Single-cell RNA-seq of NR5A1-eGFP positive cells of the developing mouse testis from E10.5 to E16.5

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