College of Fisheries, Engineering Research Center of Green development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education/National Demonstration Center for Experimental Aquaculture Education, Huazhong Agricultural University, Wuhan 430070, China.
College of Fisheries, Hubei Provincial Engineering Laboratory for Pond Aquaculture, Huazhong Agricultural University, Wuhan 430070, China.
Int J Mol Sci. 2020 Apr 19;21(8):2860. doi: 10.3390/ijms21082860.
Elongation of very long-chain fatty acids protein 6 (Elovl6) has been reported to be associated with clinical treatments of a variety of metabolic diseases. However, there is no systematic and comprehensive study to reveal the regulatory role of Elovl6 in mRNA, protein and phosphorylation levels. We established the first knock-out (KO), , in zebrafish. Compared with wild type (WT) zebrafish, KO presented significant higher whole-body lipid content and lower content of fasting blood glucose. We utilized RNA-Seq, tandem mass tag (TMT) labeling-based quantitative technology and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to perform the transcriptomic, proteomic and phosphoproteomic analyses of livers from WT and zebrafish. There were 734 differentially expressed genes (DEG) and 559 differentially expressed proteins (DEP) between and WT zebrafish, identified out of quantifiable 47251 transcripts and 5525 proteins. Meanwhile, 680 differentially expressed phosphoproteins (DEPP) with 1054 sites were found out of quantifiable 1230 proteins with 3604 sites. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) analysis of the transcriptomic and proteomic data further suggested that the abnormal lipid metabolism and glucose metabolism in KO were mainly related to fatty acid degradation and biosynthesis, glycolysis/gluconeogenesis and PPAR signaling pathway. Based on phosphoproteomic analyses, some kinases critical for lipid metabolism and glucose metabolism, including ribosomal protein S6 kinase (Rps6kb), mitogen-activated protein kinase14 (Mapk14) and V-akt murine thymoma viral oncogene homolog 2-like (Akt2l), were identified. These results allowed us to catch on the regulatory networks of on lipid and glucose metabolism in zebrafish. To our knowledge, this is the first multi-omic study of zebrafish lacking , which provides strong datasets to better understand many lipid/glucose metabolic risks posed to human health.
长链脂肪酸延长酶 6（Elovl6）已被报道与多种代谢疾病的临床治疗有关。然而，目前还没有系统和全面的研究来揭示 Elovl6 在 mRNA、蛋白质和磷酸化水平上的调节作用。我们在斑马鱼中建立了第一个基因敲除（KO）模型。与野生型（WT）斑马鱼相比，KO 表现出显著更高的全身脂肪含量和更低的空腹血糖含量。我们利用 RNA-Seq、串联质量标签（TMT）标记定量技术和液相色谱-串联质谱（LC-MS/MS）对 WT 和 KO 斑马鱼肝脏进行了转录组、蛋白质组和磷酸蛋白质组分析。在可定量的 47251 个转录本和 5525 个蛋白质中，鉴定出 734 个差异表达基因（DEG）和 559 个差异表达蛋白（DEP）。同时，在可定量的 1230 个蛋白质中有 3604 个磷酸化位点中，鉴定出 680 个差异表达磷酸蛋白（DEPP）和 1054 个磷酸化位点。转录组和蛋白质组数据的基因本体（GO）和京都基因与基因组百科全书（KEGG）分析进一步表明，KO 中异常的脂质代谢和葡萄糖代谢主要与脂肪酸降解和生物合成、糖酵解/糖异生和过氧化物酶体增殖物激活受体（PPAR）信号通路有关。基于磷酸蛋白质组学分析，鉴定出一些对脂质代谢和葡萄糖代谢至关重要的激酶，包括核糖体蛋白 S6 激酶（Rps6kb）、丝裂原活化蛋白激酶 14（Mapk14）和 V-akt 鼠胸腺瘤病毒癌基因同源物 2 样（Akt2l）。这些结果使我们能够了解 Elovl6 对斑马鱼脂质和葡萄糖代谢的调控网络。据我们所知，这是第一个缺乏 Elovl6 的斑马鱼多组学研究，为更好地理解对人类健康构成的许多脂质/葡萄糖代谢风险提供了强有力的数据集。
