ssc-miR-7134-3p regulates fat accumulation in castrated male pigs by targeting MARK4 gene.
ssc-miR-7134-3p regulates fat accumulation in castrated male pigs by targeting MARK4 gene.
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ssc-miR-7134-3p通过靶向MARK4基因调节去势雄性猪的脂肪积累
DOI：
10.7150/ijbs.17386
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发表时间：
2017
影响因子：
9.2
通讯作者：
Fang M
中科院分区：
文献类型：
--
作者：
Wang K;Li W;Bai Y;Yang W;Ling Y;Fang M
公猪去势是商业猪肉生产中减少公猪异味的常见做法，但它也会显着导致胴体脂肪堆积。我们之前的研究揭示了一个与脂肪生成有关的 miRNA 基因 ssc-miR-7134-3p。然而，ssc-miR-7134-3p 与去势引起的脂肪沉积之间的关系尚不清楚。在本研究中，我们根据生物信息学分析和双荧光素酶测定观察到 ssc-miR-7134-3p 靶向 MARK4 的编码序列 (CDS) 区域。使用沉默突变和亚克隆的实验表明，ssc-miR-7134-3p 独立结合 MARK4 CDS 中的两个相邻位点。随后，ssc-miR-7134-3p 抑制猪成纤维细胞中的 MARK4 蛋白表达，与体外证明的靶向作用一致。我们发现去势猪背部脂肪中的 MARK4 蛋白水平高于完整猪，这进一步证明 MARK4 参与脂肪沉积的调节。此外，MARK4中的一个SNP（g.2581A>G）与中国和欧洲猪群的背膘性状显着相关。综上所述，我们可以得出结论，ssc-miR-7134-3p 靶向 MARK4 基因，导致去势公猪的脂肪积累。
Castration of male pigs is a common practice used to reduce boar taint in commercial pork production, but it also significantly results in fat accumulation in carcass. Our previous study revealed a miRNA gene, ssc-miR-7134-3p that was implicated in adipogenesis. However, the relationship between ssc-miR-7134-3p and fat deposition due to castration is unknown. In the present study, we observed that ssc-miR-7134-3p targets the coding sequence (CDS) region of MARK4 based on bioinformatics analysis and dual-luciferase assays. Experiments using silent mutations and sub-cloning showed that ssc-miR-7134-3p binds independently to two adjacent sites in the MARK4 CDS. Subsequently, ssc-miR-7134-3p inhibits MARK4 protein expression in pig fibroblast cells, being consistent with the targeting demonstrated in vitro. We found higher MARK4 protein levels in the back fat of castrated pigs than in intact pigs, providing further evidence that MARK4 is involved in regulation of fat deposition. In addition, one SNP (g.2581A>G) in MARK4 was significantly associated with the back fat trait in Chinese and European pig populations. Taken together, we would conclude that ssc-miR-7134-3p targets the MARK4 gene for fat accumulation in the castrated male pigs.