Nanos2-regulated mRNAs in the cultured spermatogonial stem cells (SSCs)

Spermatogonial stem cells are foundation of spermatogenesis. In our previous study, we observed a rapid degeneration of spermatogenesis by conditional knockout (cKO) of Nanos2 in adult spermatogonia, but the underlying mechanisms are largely unknown. While, conditional overexpression (cOE) of Nanos2 also inhibited germ cell development and stayed in undifferentiation status. Here we used cultured germline stem cells (GSCs) to study the mechanisms of Nanos2 in spermatogonia. We compared Nanos2 expression in cultured GSCs with in vivo isolated Nanos2 positive cells, and found the level of Nanos2 is significantly lower in GSCs as compared to Nanos2 positive cells. Thus we generated Nano2 cOE GSCs lines. By conditional inducing expression of Nanos2 we compared gene expression profile to control GSCs. These data provide the first evidence of Nanos2 regulating genes in spermatogonial stem cells. Ectopic induction of Nanos2 in cultured GSCs. Duplicated control (Roa-CreERT2;CAG-CAT-3FlagNanos2-pA, DMSO) and experiment (Roa-CreERT2;CAG-CAT-3FlagNanos2-pA, Tamoxifen)