蛋白质组学技术对不同性别中国美利奴细毛羊(军垦型)皮肤差异蛋白的筛选

Proteomics-based screening of differentially expressed proteins in skin of Chinese merino fine sheep(JunKen type)of different gender

旨在了解性别因素对绵羊毛性状的影响。以周岁雄性和雌性中国美利奴羊(军垦型)为研究对象,利用液相色谱-串联质谱联用技术和数据非依赖性采集策略的定量蛋白质组学技术筛选皮肤组织差异表达蛋白,并对筛选获得的差异蛋白进行基因本体(gene ontology,GO)功能注释、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)代谢通路和蛋白互作分析。结果显示,共计筛选获得差异表达蛋白674种,其中,280种蛋白表达上调,394种蛋白表达下调;通过进一步分析发现,与皮肤毛囊发育及羊毛表型相关的差异蛋白有43种,上调差异蛋白30种,下调差异蛋白13种。GO注释结果显示,在分子功能方面,差异蛋白在氧结合、硫酸软骨素结合、亚铁血红素结合、谷胱甘肽过氧化物酶活性和转运活性等37个过程显著富集;在生物过程方面,差异蛋白在细胞氧化解毒作用、肌肉收缩调节、Notch信号通路、钙离子跨膜转运和谷胱甘肽新陈代谢等120个过程显著富集;在细胞组分方面,主要富集在肥大细胞颗粒、细胞核、肌质网状组织和内质网等31个过程。KEGG通路结果表明,这些差异蛋白涉及16条信号通路,其中,MAPK、P53信号通路和羊毛生长发育密切相关。蛋白质网络互作结果显示,COL1A1蛋白与MMP2、SPARC、THBS1等差异表达蛋白联系较为紧密,其可能在羊毛生长发育过程中发挥着关键作用。本研究为揭示不同性别绵羊毛性状的分子机制积累了基础数据。

The purpose of this study was to understand the effect of gender on the traits of wool.One-year-old male and female Chinese merino sheep(JunKen type)were used to screen differentially expressed proteins in skin tissues by quantitative proteomics using liquid chromatography-tandem mass spectrometry(LC-MS/MS)and data independent acquisition(DIA)strategy.This was followed by GO function annotation,KEGG metabolic pathway and protein network interaction analysis on the differential proteins obtained.The result showed that there were 674 differentially expressed proteins between male and female groups,whereas 280 proteins were up-regulated and 394 proteins were down-regulated in the male group.Through further comparison and analysis,there were 43 differentially expressed proteins related to skin hair follicle development and wool phenotype,30 up-regulated and 13 down-regulated.The GO annotation analysis of differentially expressed proteins were mainly enriched in 37 processes of molecular function such as oxygen binding,chondroitin sulfate binding,heme binding,glutathione peroxidase activity and glutathione transferase activity;120 biological processes such as cellular oxidant detoxification,regulation of muscle contraction,notch signaling pathway,calcium ion transmembrane transport and glutathione metabolism;and 31 processes related to cellular components such as mast cell granule,nucleus,sarcoplasmic reticulum and endoplasmic reticulum lumen.KEGG analysis showed that the differentially expressed proteins were involved sixteen signaling pathways,among which MAPK and p53 signaling pathways were closely related to wool growth and development.The protein network interaction analysis discovered that COL1A1 was closely related to MMP2,SPARC,THBS1 and other differentially expressed proteins,which might play a key role in wool growth and development.Thus,this study obtained basic data for revealing the molecular mechanism of sheep wool traits of different gender.