Transcriptomic profiling of young and aged spermatogonial stem cells (RNA-Seq)

Spermatogonial stem cells (SSC), the foundation of spermatogenesis and male fertility, possess lifelong self-renewal activity. Aging leads to the decline in stem cell function and increased risk of paternal age-related genetic diseases. In the present study, we performed a comparative genomic analysis of mouse SSC (Oct4-GFP+/KIT-) and differentiating progenitors (Oct4-GFP+/KIT+) isolated from young and aged testes. Our transcriptome data revealed enormous complexity of expressed coding and non-coding RNAs and alternative splicing regulation during SSC differentiation. Further comparison between young and aged SSCs suggested these differentiation programs were affected by aging. We identified aberrant expression of genes associated with meiosis and TGF-ß signaling , alteration in alternative splicing regulation and differential expression of specific lncRNAs such as Fendrr. Overall design: We separated stem cells (Oct4-GFP+/KIT-) and their immediate proliferating daughter cells (Oct4-GFP+/KIT+) using FACS from postnatal (6 days), adult (6-8 months) and aged mice (15-18 months), which were subjected to RNA-Seq analysis.