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Identification of target genes for adenohypophysis-prefer miR-7 and miR-375 in cattle

文献类型： 外文期刊

作者： Yuan, B. 1 ; Sun, G. J. 1 ; Zhang, G. L. 2 ; Wu, J. 2 ; Xu, C. 1 ; Dai, L. S. 1 ; Chen, J. 1 ; Yu, X. F. 1 ; Zhao, Z. H. 1 ; Zh 1 ;

作者机构： 1.Jilin Univ, Coll Anim Sci, Changchun 130023, Peoples R China

2.Jilin Acad Agr Sci, Branch Anim Husb, Gongzhuling, Peoples R China

关键词： Prostaglandin F2 receptor negative regulator;miR-375;miR-7;Bone morphogenetic protein receptor type II;Bovine adenohypophysis

期刊名称：GENETICS AND MOLECULAR RESEARCH （ 影响因子：0.764； 五年影响因子：0.912 ）

ISSN： 1676-5680

年卷期： 2015 年 14 卷 3 期

页码：

收录情况： SCI

摘要： In this study, expression levels of miRNAs (miRNAs), miR-375 and miR-7, were detected in different tissues of cattle to determine whether adenohypophysis-prefer or exclusively expressed miRNAs, and target genes could be predicted by TargetScan, RNA22, and other software. Target genes related to pituitary function or reproductive traits were identified using a dual-luciferase assay. miR-375 and miR-7 were expressed differently in various tissues. miR-375 and miR-7 showed higher expression in the adenohypophysis, and there was a significant difference compared with expression in other tissues (P < 0.01). The binding sites for miR-7 were the mRNAs of bone morphogenetic protein receptor type II (BMPR2), prostaglandin F2 receptor negative regulator, gonadotropin-releasing hormone receptor, follicle-stimulating hormone beta, somatostatin receptor 1, and interleukin-1 beta by bioinformatic analysis; similarly, the mRNAs of BMPR2 and leptin contained binding sites for miR-375, suggesting that these genes are affected by miR-7 or miR-375. Dual-luciferase reporter assays showed that miR-7 regulated prostaglandin F2 receptor negative regulator expression, while miR-375 regulated BMPR2 expression. The mutated plasmid and miRNA mimics were used to co-transfect NIH3T3 cells; luciferase reporter assays showed that the inhibition of luciferase activity in the wild-type cells dramatically decreased from 75 to 26% with a 3-5-nucleotide mismatch mutation into the seed region of miR-7. miR-375 had nearly lost the ability to inhibit luciferase activity, suggesting that GTCTTCC is the site of interaction between miR-7 and the prostaglandin F2 receptor negative regulator sequence and that GAACAAA is the site of interaction between miR-375 and the BMPR2 sequence.

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