Zhao Qian, Kang Ye, Wang Hong-Yang, Guan Wei-Jun, Li Xiang-Chen, Jiang Lin, He Xiao-Hong, Pu Ya-Bin, Han Jian-Lin, Ma Yue-Hui, Zhao Qian-Jun
Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
CAAS-ILRI Joint Laboratory on Livestock and Forage Genetic Resources, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Sci Rep. 2016 Jul 25;6:30281. doi: 10.1038/srep30281.
MicroRNAs (miRNAs) are evolutionarily conserved, small, non-coding RNAs that have emerged as key regulators of myogenesis. Here, we examined the miRNA expression profiles of developing sheep skeletal muscle using a deep sequencing approach. We detected 2,396 miRNAs in the sheep skeletal muscle tissues. Of these, miR-192 was found to be up-regulated in prenatal skeletal muscle, but was down-regulated postnatally. MiR-192 expression also decreased during the myogenic differentiation of sheep satellite cells (SCs). MiR-192 overexpression significantly attenuated SCs myogenic differentiation but promoted SCs proliferation, whereas miR-192 inhibition enhanced SCs differentiation but suppressed SCs proliferation. We found that miR-192 targeted retinoblastoma 1 (RB1), a known regulator of myogenesis. Furthermore, knockdown of RB1 in cultured cells significantly inhibited SCs myogenic differentiation but accelerated SCs proliferation, confirming the role of RB1 in myogenesis. Taken together, our findings enrich the ovine miRNA database, and outline the miRNA transcriptome of sheep during skeletal muscle development. Moreover, we show that miR-192 affects SCs proliferation and myogenic differentiation via down-regulation of RB1.
微小RNA（miRNA）是进化上保守的小非编码RNA，已成为肌生成的关键调节因子。在此，我们使用深度测序方法研究了发育中的绵羊骨骼肌的miRNA表达谱。我们在绵羊骨骼肌组织中检测到2396种miRNA。其中，miR-192在产前骨骼肌中上调，但在出生后下调。miR-192的表达在绵羊卫星细胞（SCs）的肌源性分化过程中也降低。miR-192过表达显著减弱SCs的肌源性分化，但促进SCs增殖，而miR-192抑制则增强SCs分化但抑制SCs增殖。我们发现miR-192靶向视网膜母细胞瘤1（RB1），一种已知的肌生成调节因子。此外，在培养细胞中敲低RB1显著抑制SCs的肌源性分化但加速SCs增殖，证实了RB1在肌生成中的作用。综上所述，我们的研究结果丰富了绵羊miRNA数据库，并勾勒出绵羊骨骼肌发育过程中的miRNA转录组。此外，我们表明miR-192通过下调RB1影响SCs增殖和肌源性分化。
