miR-204 Regulates the Proliferation of Dairy Goat Spermatogonial Stem Cells via Targeting to Sirt1

• Published in: Rejuvenation research Vol. 19; no. 2; p. 120
• Main Authors: Niu, Bowen, Wu, Jiang, Mu, Hailong, Li, Bo, Wu, Chongyang, He, Xin, Bai, Chunling, Li, Guangpeng, Hua, Jinlian
• Format: Journal Article
• Language: English
• Published: United States 01.04.2016
• Subjects: 3' Untranslated Regions - genetics; Animals; Antigens, CD - metabolism; Base Sequence; Cell Proliferation; Cell Self Renewal; Flow Cytometry; Gene Expression Regulation; Genes, Reporter; Genetic Vectors - metabolism; Goats - genetics; Luciferases - metabolism; Magnetic Phenomena; Male; MicroRNAs - genetics; MicroRNAs - metabolism; Models, Biological; Sirtuin 1 - genetics; Sirtuin 1 - metabolism; Spermatogonia - cytology; Spermatogonia - metabolism; Stem Cells - cytology; Stem Cells - metabolism
• ISSN: 1557-8577
• DOI: 10.1089/rej.2015.1719

The regulation of spermatogonial stem cell (SSC) proliferation and self-renewal is a complex process. Several studies on the microRNA regulation of mammalian spermatogenesis have been reported. Here, we predicted miRNA targeting of Sirt1, and a dual luciferase experiment confirmed that miR-204 interacted with the Sirt1 3'-untranslated region (3'-UTR). The expression of miR-204 and Sirt1 in dairy goat testicles was investigated, and the results showed that the expression pattern of Sirt1 was similar to that of miR-204 in the temporal-spatial distribution. The over-expression of Sirt1 in goat SSCs can promote SSCs' self-renewal gene expression and cell proliferation. Furthermore, miRNA sequencing results showed that Sirt1 had a higher expression level in dairy goat CD49f(+) and CD90(+) SSCs, but the expression level of miR-204 was lower. In an in vitro assay, Sirt1 was significantly down-regulated in dairy goat SSCs when transfected with miR-204 mimics, indicating that Sirt1 was a target of miR-204 in the dairy goat. On the basis of the results of RT-qPCR, fluorescence-activated cell sorting (FACS), and western blotting, we found that the over-expression of Sirt1 in goat SSCs can promote cellular proliferation and change self-renewal and pluripotent gene expression. Thus, miR-204 was involved in the regulation of dairy goat SSCs proliferation via Sirt1.