Zuo Jianjun, Wu Fan, Liu Yihua, Xiao Juan, Xu Mei, Yu Qinping, Xia Minhao, He Xiaojun, Zou Shigeng, Tan Huize, Feng Dingyuan
College of Animal Science of South China Agricultural University, Guangzhou, 510642, China.
Production Technology Department of WENs Group, Xinxing, 527400, China.
PLoS One. 2015 Apr 27;10(4):e0124873. doi: 10.1371/journal.pone.0124873. eCollection 2015.
Porcine skeletal muscle fibres are classified based on their different physiological and biochemical properties. Muscle fibre phenotype is regulated by several independent signalling pathways, including the mitogen-activated protein kinase (MAPK), nuclear factor of activated T cells (NFAT), myocyte enhancer factor 2 (MEF2) and peroxisome proliferator-activated receptor (PPAR) signalling pathways. MicroRNAs are non-coding small RNAs that regulate many biological processes. However, their function in muscle fibre type regulation remains unclear. The aim of our study was to identify miRNAs that regulate muscle fibre type during porcine growth to help understand the miRNA regulation mechanism of fibre differentiation. We performed Solexa/Illumina deep sequencing for the microRNAome during 3 muscle growth stages (63, 98 and 161 d). In this study, 271 mature miRNAs and 243 pre-miRNAs were identified. We detected 472 novel miRNAs in the muscle samples. Among the mature miRNAs, there are 23 highest expression miRNAs (over 10,000 RPM), account for 85.3% of the total counts of mature miRNAs., including 10 (43.5%) muscle-related miRNAs (ssc-miR-133a-3p, ssc-miR-486, ssc-miR-1, ssc-miR-143-3p, ssc-miR-30a-5p, ssc-miR-181a, ssc-miR-148a-3p, ssc-miR-92a, ssc-miR-21, ssc-miR-126-5p). Particularly, both ssc-miR-1 and ssc-miR-133 belong to the MyomiRs, which control muscle myosin content, myofibre identity and muscle performance. The involvement of these miRNAs in muscle fibre phenotype provides new insight into the mechanism of muscle fibre regulation underlying muscle development. Furthermore, we performed cell transfection experiment. Overexpression/inhibition of ssc-miR-143-3p in porcine skeletal muscle satellite cell induced an/a increase/reduction of the slow muscle fibre gene and protein (MYH7), indicating that miR-143 activity regulated muscle fibre differentiate in skeletal muscle. And it regulate MYH7 through the HDAC4-MEF2 pathway.
猪骨骼肌纤维根据其不同的生理和生化特性进行分类。肌纤维表型受多种独立信号通路调控，包括丝裂原活化蛋白激酶（MAPK）、活化T细胞核因子（NFAT）、肌细胞增强因子2（MEF2）和过氧化物酶体增殖物激活受体（PPAR）信号通路。微小RNA是一类非编码小RNA，可调控多种生物学过程。然而，它们在肌纤维类型调控中的作用仍不清楚。我们研究的目的是鉴定在猪生长过程中调控肌纤维类型的微小RNA，以帮助理解纤维分化的微小RNA调控机制。我们对3个肌肉生长阶段（63、98和161日龄）的微小RNA组进行了Solexa/Illumina深度测序。在本研究中，鉴定出271个成熟微小RNA和243个前体微小RNA。我们在肌肉样本中检测到472个新的微小RNA。在成熟微小RNA中，有23个表达量最高的微小RNA（超过10,000 RPM），占成熟微小RNA总数的85.3%，其中包括10个（43.5%）与肌肉相关的微小RNA（ssc-miR-133a-3p、ssc-miR-486、ssc-miR-1、ssc-miR-143-3p、ssc-miR-30a-5p、ssc-miR-181a、ssc-miR-148a-3p、ssc-miR-92a、ssc-miR-21、ssc-miR-126-5p）。特别地，ssc-miR-1和ssc-miR-133都属于肌微小RNA，可控制肌肉肌球蛋白含量、肌纤维特性和肌肉性能。这些微小RNA参与肌纤维表型的调控，为肌肉发育过程中肌纤维调控机制提供了新的见解。此外，我们进行了细胞转染实验。在猪骨骼肌卫星细胞中过表达/抑制ssc-miR-143-3p可诱导慢肌纤维基因和蛋白（MYH7）的增加/减少，表明miR-143活性在骨骼肌中调控肌纤维分化。并且它通过HDAC4-MEF2途径调控MYH7。
