基于iTRAQ蛋白组学技术挖掘水貂生长调控关键蛋白

Excavating key proteins regulating growth in mink based on iTRAQ proteomics technology

为了分析水貂在不同日龄胸肌组织蛋白质表达模式,探究水貂生长发育调控机制,试验选取相同条件下饲养的健康雄性银蓝水貂9只,分别在45日龄、90日龄、120日龄时各宰杀3只取胸肌组织,利用iTRAQ定量蛋白组学技术构建银蓝水貂蛋白质表达图谱,筛选显著差异表达蛋白,并进行GO功能注释、KEGG信号通路富集、时间趋势聚类分析。结果表明:在银蓝水貂胸肌组织中共鉴定到2922种蛋白质,其中189种在水貂生长过程中发生了显著差异表达(差异倍数≥1.2或≤0.83,P<0.05),有25种差异表达蛋白与肌肉发育相关。差异表达蛋白主要参与了核糖体生物合成、蛋白靶向内质网、翻译、肌动蛋白介导的细胞收缩、肌丝滑动、ATP酶活性调控等生物学过程;富集到了核糖体、翻译、氧化磷酸化、能量代谢、心肌收缩等信号通路中。差异表达蛋白在Profile 2与Profile 5两种表达模式显著富集,Profile 2模式中的差异表达蛋白在90日龄表达量显著下降,主要参与了翻译、蛋白质代谢过程、核糖体生物合成;Profile 5模式中的差异表达蛋白在90日龄表达量最高,主要参与了骨骼肌系统、肌细胞发育、氧化磷酸化。说明这些差异表达蛋白可以作为水貂生长发育相关的候选蛋白,用于水貂生长发育分子调控机制的阐明及大型优质水貂品种的培育。

In order to analyze the protein expression pattern of the pectoral muscle tissue of mink at different ages,and to investigate the regulatory mechanism of mink growth and development,in the experiment,nine healthy male Silver Blue mink were selected under the same feeding conditions;their pectoral muscle tissues were taken at 45,90 and 120 days of age;iTRAQ quantitative proteomics technology was used to construct protein expression profiles of Silver Blue mink for screening for significantly differentially expressed proteins;and GO function enrichment,KEGG signaling pathway enrichment,and time-trend clustering analysis were performed.The results showed that a total of 2,922 proteins were identified in the pectoral muscle tissues of Silver Blue mink;among them,189 were significantly differentially expressed during the growth of mink(multiplicity of difference≥1.2 or≤0.83,P<0.05);25 differentially expressed proteins were related to muscle development.The differentially expressed proteins were mainly involved in biological processes such as ribosome biosynthesis,protein targeting to endoplasmic reticulum,translation,actin-mediated cell contraction,myofilament sliding,and regulation of ATPase activity;signaling pathways were enriched such as ribosome,translation,oxidative phosphorylation,energy metabolism,and myocardial contraction;Dynamic analysis of differentially expressed protein expression showed that the expression patterns of Profile 2 and Profile 5 were significantly enriched,and the expression of differentially expressed proteins in Profile 2 pattern was significantly decreased at 90 days of age,which were mainly involved in translation,protein metabolic process and ribosome biosynthesis.Differential expression proteins in the Profile 5 pattern were most highly expressed at 90 days of age,which were related to skeletal muscle system,myocyte development and oxidative phosphorylation.The results suggested that these differentially expressed proteins could be used as candidate proteins related to mink growth and development,and be used for the elucidation of the molecular regulatory mechanism of mink growth and development and for the cultivation of large-scale and high-quality mink breeds.