title: miR-204 Regulates the Proliferation of Dairy Goat Spermatogonial Stem Cells via Targeting to Sirt1.
abstract: 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.
authors.affiliation: 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University , Yangling, Shaanxi, China .
authors.forename: Bowen
authors.initials: B
authors.lastname: Niu
authors.affiliation: 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University , Yangling, Shaanxi, China .|2 College of Agriculture, Guangdong Ocean University , Zhanjiang, China .
authors.forename: Jiang
authors.initials: J
authors.lastname: Wu
authors.affiliation: 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University , Yangling, Shaanxi, China .
authors.forename: Hailong
authors.initials: H
authors.lastname: Mu
authors.affiliation: 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University , Yangling, Shaanxi, China .
authors.forename: Bo
authors.initials: B
authors.lastname: Li
authors.affiliation: 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University , Yangling, Shaanxi, China .
authors.forename: Chongyang
authors.initials: C
authors.lastname: Wu
authors.affiliation: 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University , Yangling, Shaanxi, China .
authors.forename: Xin
authors.initials: X
authors.lastname: He
authors.affiliation: 3 Key Laboratory for Mammalian Reproductive Biology and Biotechnology, Ministry of Education, Inner Mongolia University , Hohhot, China .
authors.forename: Chunling
authors.initials: C
authors.lastname: Bai
authors.affiliation: 3 Key Laboratory for Mammalian Reproductive Biology and Biotechnology, Ministry of Education, Inner Mongolia University , Hohhot, China .
authors.forename: Guangpeng
authors.initials: G
authors.lastname: Li
authors.affiliation: 1 College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University , Yangling, Shaanxi, China .
authors.forename: Jinlian
authors.initials: J
authors.lastname: Hua
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001483:Base Sequence; D049109:Cell Proliferation; D000066673:Cell Self Renewal; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000378:metabolism; D006041:Goats / Q000235:genetics; D008156:Luciferases / Q000378:metabolism; D060328:Magnetic Phenomena; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D056564:Sirtuin 1 / Q000235:genetics / Q000378:metabolism*; D013093:Spermatogonia / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; D015703:Antigens, CD; D035683:MicroRNAs; D008156:Luciferases; D056564:Sirtuin 1