金茅黑鸡miR-206表达及靶基因预测分析

doi:10.16431/j.cnki.1671-7236.2020.02.022

摘要/Abstract

摘要： 为了研究miR-206的组织表达分布及其在鸡骨骼肌生长发育中的表达规律，预测其可能的作用机制，本研究采集4周龄金茅黑鸡的心脏、肝脏、脾脏、肺脏、肾脏、胸肌、腿肌和腹脂8种组织及2、6、10、14、16周龄胸肌和腿肌组织，利用实时荧光定量PCR技术检测miR-206在8种组织及不同周龄胸肌和腿肌中的表达，利用生物信息学方法预测其靶基因并进行功能注释。结果显示，miR-206在公、母鸡胸肌和腿肌中表达量均极显著高于其他组织（P < 0.01），在腹脂、心脏、肝脏、脾脏、肺脏和肾脏中低表达，为鸡骨骼肌特异性表达miRNA；miR-206均在公、母鸡腿肌和胸肌组织生长早期（2周龄）表达最高，之后表达量逐渐下降。靶基因预测和功能分析发现，miR-206共有356个靶基因，其中21个与肌肉生成相关，多个靶基因已知参与调控肌肉生成，包括配对框7（paired box 7，Pax7）、胰岛素样生长因子1（insulin-like growth factor 1，IGF1）、脑衍生神经营养因子1（brain-derived neurotrophic factor 1，BDNF1）、视黄酸受体（retinoic acid receptor beta，RARB）和卷曲类受体7（frizzled class receptor 7，FZD7）基因。此外，发现miR-206靶基因富集到多条已知参与调控骨骼肌生成的信号通路，包括MAPK、Wnt、肌动蛋白骨架调控和黏着斑激酶信号通路。结合表达数据推测，miR-206可能通过靶向调控Pax7、IGF1、BDNF1、RARB、FZD7基因和MAPK、Wnt等信号通路参与调控鸡肌肉生成。本研究揭示了金茅黑鸡miR-206的组织表达分布及其在鸡骨骼肌生长过程中的表达规律，并预测了其调控骨骼肌生成的可能作用机理，为进一步研究miR-206在鸡肌肉生长发育中的作用机理和表达调控奠定了基础。

关键词: miR-206; 金茅黑鸡; 组织表达分布; 表达规律; 靶基因

Abstract: The objective of this study was to study the tissue expression distribution of miR-206 and its expression pattern in skeletal muscle growth,and predict its possible mechanism of action.Eight tissues of heart,liver,spleen,lung,kidney,breast muscle,leg muscle and abdominal fat were collected from 4-week-old Jinmao Black chickens.Breast muscle and leg muscle were collected from 2,6,10,14 and 16-week-old Jinmao Black chickens.The expression of miR-206 in eight tissues and skeletal muscle at different growth stages using Real-time PCR.The target genes of miR-206 were predicted and conducted to function annotation by bioinformatics methods.The results showed that the expression level of miR-206 in breast muscle and leg muscle was extremely significantly higher than that in other tissues (P < 0.01).miR-206 was lowly expressed in abdominal fat,heart,liver,spleen,lung and kidney,and specifically expressed in skeletal muscle.The expression pattern showed that the expression of miR-206 was the highest in leg muscle and breast muscle at 2-weeks-old,and showed a downward trend in general.Target gene prediction and functional analysis revealed that miR-206 had 356 target genes,21 of which were involved in myogenesis.Multiple target genes were known to be involved in the regulation of myogenesis,including paired box 7 (Pax7),insulin-like growth factor 1 (IGF1),brain-derived neurotrophic factor 1 (BDNF1),retinoic acid receptor beta (RARB) and frizzled class receptor 7 (FZD7) genes.Pathway analysis showed that four pathways involved in myogenesis were significantly enriched,including MAPK signaling pathway,Wnt signaling pathway,regualtion of actin cytoskeleton and focal adhesion pathway.Combined with the expression results,we speculated that miR-206 might participate in the regulation of chicken muscle myogenesis by targeting Pax7、IGF1、BDNF1、RARB, FZD7 genes and MAPK,Wnt,regulation of actin cytoskeleton and focal adhesion pathways.This study revealed the tissue distribution and expression pattern of miR-206 in the growth of chicken and predicted its possible function and mechanism in the regulation of skeletal muscle myogenesis,which was the important foundation and basis for further study of the mechanism of action and expression regulation of miR-206.

Key words: miR-206; Jinmao Black chickens; tissue expression distribution; expression pattern; target gene

中图分类号:

Q786

黄金星, 陈兰, 张涛, 陈福祥, 吴鹏飞, 冒静, 巩用双, 谢恺舟, 王金玉, 戴国俊. 金茅黑鸡miR-206表达及靶基因预测分析[J]. 中国畜牧兽医, 2020, 47(2): 506-513.

HUANG Jinxing, CHEN Lan, ZHANG Tao, CHEN Fuxiang, WU Pengfei, MAO Jing, GONG Yongshuang, XIE Kaizhou, WANG Jinyu, DAI Guojun. A Expression Analysis and Target Gene Prediction of miR-206 in Jinmao Black Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(2): 506-513.

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