miR-19b-3p induces cell proliferation and reduces heterochromatin-mediated senescence through PLZF in goat male germline stem cells.

Promyelocytic leukemia zinc finger PLZF, known as ZBTB16 or ZFP145 is a critical zinc finger protein of male germline stem cells (mGSCs), it's an essential transcriptional factor for goat testis development and spermatogenesis. Loss of PLZF results in progressive depletion of SSCs after the first wave of spermatogenesis leading to eventual spermatogenic arrest, apparently the result of a shift in the balance in SSC fate away from self-renewal and toward differentiation. Cumulating evidences have demonstrated that microRNAs are expressed in a cell-specific or stage-specific manner during spermatogenesis and acts as regulators on specific makers such as Stra8, ETV5, and PLZF. However, the post transcriptional function of PLZF still poorly elucidate in mGSCs. Bioinformatic analysis and dual luciferase reporter assay showed that miR-19b-3p binds the 3'UTR of PLZF, suggesting that PLZF is a direct target of miR-19b-3p. The profile of miR-19b-3p and PLZF analyzed in dairy goat testis at different age showed that miR-19b-3p was significantly up-regulated in goat testis at 1, 3, 6 months and downregulated at 12, 18, and 24 months which was inversely correlated with PLZF in the same testis. Focusing on the role of miR-19b-3p, we found that miR-19b-3p changes c-KIT and mTOR signaling through PLZF to promote proliferation in goat nGSCs and infertile mice testes. Over-expression of PLZF significantly reversed miR-19b-3p-mediated proliferation in mice testes. We found also that miR-19b-3p reduced heterochromatin-mediated senescence through PLZF localized on HP1α. Taken together, our findings indicate that miR-19b-3p promotes proliferation and reduces heterochromatin-mediated senescence through PLZF in mGSCs.