Transcriptomic dynamics and cell-to-cell communication during the transition of prospermatogonia to spermatogonia revealed at single-cell resolution

Spermatogonia are essential for the continual production of sperm and regeneration of the entire spermatogenic lineage after injury. In mammals, spermatogonia are formed in the neonatal testis from prospermatogonia (also termed gonocytes), which are established from primordial germ cells during fetal development. Currently, the molecular regulation of the prospermatogonial to spermatogonia transition is not fully understood.

Together, these findings provide a crucial dataset for dissecting key genes that direct the establishment of the foundational spermatogonial pool and the fate transitions of different somatic cell lineages in the testis during fetal and neonatal periods of development.

In this study, we examined the gene expression patterns of prospermatogonia, spermatogonia and testicular somatic cells at 4 different stages, including embryonic day (E) 12.5, E17.5 and postnatal days (P) 1 and 6, using single-cell RNA sequencing (scRNA-seq). We identified 5 different molecular states in the prospermogonial population and revealed gene expression dynamics in corresponding testicular somatic cells. Specifically, we found that prospermatogonia mainly receive signals, while Leydig cells and peritubular myoid cells are the mediators for transmitting signals, indicating their potential roles in regulating the development and differentiation of prospermatogonia. Transcription regulon analyses revealed the involvement of basic helix-loop-helix (bHLH) transcription factors in directing prospermogonial fate decisions. We then disrupted this transcription network by ectopic expression of inhibitor of differentiation 2 (Id2), which is a negative regulator of bHLH transcription factors. The overexpression of Id2 in prospermatogonia caused severe defects in the progression of prospermatogonia to spermatogonia.