Sun Wen-Kui, Li Yanyue, Cheng Chi, Chen Yi-Hui, Zeng Kai, Chen Xiaohui, Gu Yiren, Liu Rui, Lv Xuebin, Gao Rong
Key Laboratory of Bio-resource and Eco-Environment of Education Ministry, Key Laboratory of Animal Disease Prevention and Food Safety of Sichuan Province, College of Life Sciences, Sichuan University, Wangjiang Road 29, Chengdu, 610064, Sichuan, China.
School of Laboratory Medicine, Chengdu Medical College, Chengdu, 610500, Sichuan, China.
Genes Genomics. 2018 Sep;40(9):937-943. doi: 10.1007/s13258-018-0696-y. Epub 2018 May 14.
MiRNAs regulate the expression of target genes in diverse cellular processes and hence play important roles in different physiological processes, yet little is known about the stomach microRNAome (miRNAome) of the Tibetan pig. The objective of this experiment was to investigate differentially expressed stomach miRNAs participating in digestion. Firstly, we isolated total RNA by Trizol reagent from three Tibetan and three Yorkshire purebred pigs stomach samples at 90-day-old. Secondly, a comprehensive analysis of Tibetan and Yorkshire pig stomach miRNAomes was performed by small RNA sequencing in the Illumina HiSeq 2000 system. Finally, SYBR Green Real-time RT-PCR was performed to validate the differentially expressed miRNAs. We identified 318 unique miRNAs, 260 were co-expressed in both libraries, 17 and 31 miRNAs were specifically expressed in Tibetan and Yorkshire pigs respectively. Fifty six differentially expressed miRNAs were identified by the identifying differentially expressed genes 6 (IDEG6). Kyoto encyclopedia of genes and genomes analysis revealed that some of the differentially expressed miRNAs were associated with protein and fat digestion. Two differentially expressed miRNAs (miR-214-3p and ssc-un39) participating in the digestion of lipid were identified. Additionally, qRT-PCR results suggested that a higher expression of miR-214-3p in the Tibetan pig stomach could lead to relatively lower expression of calcium-dependent phospholipase A2, which is an enzyme important for the digestion of glycerol phospholipid. This study has delineated the different stomach miRNAs expression patterns of Tibetan and Yorkshire pigs, which would help explain the regulatory mechanisms of miRNAs in digestion of Tibetan pigs, and contribute to utilize a the unique digestion merits of Tibetan pig in future porcine hybridization breeding.
微小RNA（miRNA）在多种细胞过程中调节靶基因的表达，因此在不同的生理过程中发挥重要作用，但关于藏猪胃微小RNA组（miRNA组）的了解却很少。本实验的目的是研究参与消化的差异表达胃miRNA。首先，我们使用Trizol试剂从三头90日龄的藏猪和三头约克夏纯种猪的胃样本中分离总RNA。其次，在Illumina HiSeq 2000系统中通过小RNA测序对藏猪和约克夏猪的胃miRNA组进行了全面分析。最后，进行SYBR Green实时逆转录PCR以验证差异表达的miRNA。我们鉴定出318个独特的miRNA，其中260个在两个文库中共同表达，分别有17个和31个miRNA在藏猪和约克夏猪中特异性表达。通过鉴定差异表达基因6（IDEG6）鉴定出56个差异表达的miRNA。京都基因与基因组百科全书分析表明，一些差异表达的miRNA与蛋白质和脂肪消化有关。鉴定出两个参与脂质消化的差异表达miRNA（miR-214-3p和ssc-un39）。此外，qRT-PCR结果表明，藏猪胃中miR-214-3p的较高表达可能导致钙依赖性磷脂酶A2的表达相对较低，钙依赖性磷脂酶A2是一种对甘油磷脂消化很重要的酶。本研究描绘了藏猪和约克夏猪不同的胃miRNA表达模式，这将有助于解释miRNA在藏猪消化中的调控机制，并有助于在未来的猪杂交育种中利用藏猪独特的消化优势。
