bta-miR-377靶向调控牛下丘脑CART基因表达的研究

doi:10.16431/j.cnki.1671-7236.2022.09.004

Abstract

Abstract: 【Objective】 This study was aimed to analyze the interaction between bta-miR-377 and cocaine and amphetamine regulated transcript peptide(CART), a key regulatory factor of bovine follicle development, and to clarify the mechanism of bta-miR-377 on the expression of CART.【Method】 In this study, the binding sites of bta-miR-377 and bovine CART gene mRNA were predicted by bioinformatics, and the sequence conservation of miR-377 in different species was analyzed.The target relationship between bta-miR-377 and CART gene was verified by double luciferase reporter gene method.Three healthy Simmental cows were selected to collect their hypothalamic tissues and analyze the endogenous expression of bta-miR-377 and CART gene in the hypothalamus.PC12 and 293T cells were used as models for cell function verification.The abundance of CART gene mRNA in PC12 cells and CART protein in 293T cells transfected with bta-miR-377 mimics and NC mimics were detected by Real-time quantitative PCR and Western blotting, respectively.【Result】 bta-miR-377 had binding site to CART 3'-UTR and extremely significantly down-regulated the relative fluorescence activity of wild-type recombinant double fluorescent plasmid (P<0.01).The binding site sequences between the bta-miR-377 and CART 3'-UTR were highly conserved in different species.Lower sequence conservation in the miR-377 seed region occurred between cows and rats.Both of bta-miR-377 and CART were detected in bovine hypothalamus.Overexpressing bta-miR-377 mimics could significantly down-regulated the expression of CART gene mRNA and CART protein (P<0.01 or P<0.05).There was a negative correlation between bta-miR-377 and CART expression.【Conclusion】 Both bta-miR-377 and CART were expressed in bovine hypothalamic and bta-miR-377 inhibits CART expression at level of mRNA by specifically binding to CART 3'-UTR in PC12 cells.

Key words: cow; hypothalamus; CART; bta-miR-377

CLC Number:

S823.2

REN Jing, HAO Qinqin, CHENG Junli, ZHU Zhiwei, XU Dongmei, JIA Xuechun, LI Pengfei. Expression Analysis of bta-miR-377 Regulate CART Gene in Bovine Hypothalamus[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3301-3309.

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Expression Analysis of bta-miR-377 Regulate CART Gene in Bovine Hypothalamus

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