title: miR-361-3p regulates FSH by targeting FSHB in a porcine anterior pituitary cell model.
abstract: FSH plays an essential role in processes involved in human reproduction, including spermatogenesis and the ovarian cycle. While the transcriptional regulatory mechanisms underlying its synthesis and secretion have been extensively studied, little is known about its posttranscriptional regulation. A bioinformatics analysis from our group indicated that a microRNA (miRNA; miR-361-3p) could regulate FSH secretion by potentially targeting the FSHB subunit. Herein, we sought to confirm these findings by investigating the miR-361-3p-mediated regulation of FSH production in primary pig anterior pituitary cells. Gonadotropin-releasing hormone (GnRH) treatment resulted in an increase in FSHB synthesis at both the mRNA, protein/hormone level, along with a significant decrease in miR-361-3p and its precursor (pre-miR-361) levels in time- and dose-dependent manner. Using the Dual-Luciferase Assay, we confirmed that miR-361-3p directly targets FSHB. Additionally, overexpression of miR-361-3p using mimics significantly decreased the FSHB production at both the mRNA and protein levels, with a reduction in both protein synthesis and secretion. Conversely, both synthesis and secretion were significantly increased following miR-361-3p blockade. To confirm that miR-361-3p targets FSHB, we designed FSH-targeted siRNAs, and co-transfected anterior pituitary cells with both the siRNA and miR-361-3p inhibitors. Our results indicated that the siRNA blocked the miR-361-3p inhibitor-mediated upregulation of FSH, while no significant effect on non-target expression. Taken together, our results demonstrate that miR-361-3p negatively regulates FSH synthesis and secretion by targeting FSHB, which provides more functional evidence that a miRNA is involved in the direct regulation of FSH.
authors.forename: Rui-Song
authors.initials: RS
authors.lastname: Ye
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Chao-Yun
authors.initials: CY
authors.lastname: Li
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Qi-En
authors.initials: QE
authors.lastname: Qi
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Chen
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Cheng
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Song-Bo
authors.initials: SB
authors.lastname: Wang
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Shu
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Li-Na
authors.initials: LN
authors.lastname: Wang
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Xiao-Tong
authors.initials: XT
authors.lastname: Zhu
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
authors.forename: Qing-Yan
authors.initials: QY
authors.lastname: Jiang
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China xqy0228@163.com Zhangyl@scau.edu.cn.
authors.forename: Qian-Yun
authors.initials: QY
authors.lastname: Xi
authors.affiliation: Chinese National Engineering Research Center for Breeding Swine IndustrySCAU-Alltech Research Joint Alliance, Guandong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China xqy0228@163.com Zhangyl@scau.edu.cn.
authors.forename: Yong-Liang
authors.initials: YL
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D005640:Follicle Stimulating Hormone / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008954:Models, Biological; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011962:Receptors, FSH / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D015854:Up-Regulation
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D011962:Receptors, FSH; D007987:Gonadotropin-Releasing Hormone; D005640:Follicle Stimulating Hormone