miR-31 Regulates Spermatogonial Stem Cells Meiosis via Targeting Stra8

4 years ago

miR-31 Regulates Spermatogonial Stem Cells Meiosis via Targeting Stra8

Yingjie Wang, Bichun Li, Jing Jin, Yulin Bi, Ya-ni Zhang, Wenhui Zhang, Liangliang Zhang, Qisheng Zuo

Stra8 (stimulated by retinoic acid gene 8) is a specific gene that is expressed in mammalian germ cells during transition from mitosis to meiosis and plays a key role in the initiation of meiosis in mammals and birds. So, the evaluation of the Stra8 pathway in cSSCs may provide a deeper insight into mammalian spermatogenesis. miRNA was also an important regulating factor for meiosis of SSCs. However, there is currently no data indicating that miRNA regulate the meiosis of SSCs via Stra8. Here, we predicted the prospective miRNA targeting to Stra8 using the online Bioinformatics database-Targetscan, and performed an analysis of the dual-luciferase recombinant vector, pGL3-CMV-LUC-MCS-Stra8-3′UTR. miR-31 mimics (miR-31m), miR-31 inhibitors (miR-31i), Control (NC, scrambled oligonucleotides transfection) were transfected into cSSCs; Stra8 and miRNA were analyzed by RT-qPCR, immunofluorescence, and Western blot. The detection of haploid was conducted by flow cytometry. The results showed that miR-31 regulates meiosis of cSSCs via targeting Stra8 in vitro and in vivo. Our study identifies a new regulatory pathway that miR-31 targets Stra8 and inhibits spermatogenesis. J. Cell. Biochem. 118: 4844–4853, 2017. © 2017 Wiley Periodicals, Inc. In this work, we predicted the prospective miRNA targeting to Stra8 and performed an analysis of the dual-luciferase recombinant vector, pGL3-CMV-LUC-MCS-Stra8-3′UTR. Stra8 and miRNA were analyzed by RT-qPCR, immunofluorescence, and Western blot; the detection of haploid was conducted by flow cytometry. Our results indicated that miR-31 regulates meiosis of cSSCs via targeting Stra8 in vitro and in vivo.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/jcb.26159