大白母猪发情期前后唾液蛋白组学分析

Salivary proteomics analysis of large white sows before and after estrus

为解决养猪生产中传统发情鉴定法工作量大、准确率低等问题,本研究以经产大白母猪为研究对象,将发情周期分为发情前3天(PE),发情期(E)和发情后3天(AE)3个阶段,分别收集口腔中唾液,提取唾液蛋白,使用Lable-free定量蛋白组学技术对3个阶段的唾液蛋白组进行质谱检测,并通过蛋白质组学定性定量软件Maxquant进行分析。结果显示:1)3个阶段共鉴定到405个蛋白质,包括149个共有蛋白和160个特异蛋白,其中有45个共有蛋白差异达到显著水平(P<0.05),PE、E和AE 3个阶段分别有54、25和81个特异蛋白;2)对差异蛋白进行GO和KEGG富集分析,其参与主要的代谢途径包括胰高血糖素信号通路、丙酮酸代谢,内肽酶抑制剂活性和蛋白质水解调节等;3)进一步对筛选到的蛋白进行互作分析,P79263、F1SFI7、Q29549、F1SS24、Q9GMA6和Q2EN74蛋白间存在互作关系,其中F1SS24、Q9GMA6和P79263属于PE期,F1SFI7、Q29549和Q2EN74为E期蛋白。综上,本研究通过对大白母猪发情期不同阶段的唾液蛋白组进行分析,筛选到多个目标差异/特异蛋白,本研究为揭示母猪发情周期唾液蛋白含量变化提供了参考依据,并为下一步发情鉴定试剂盒/试纸条的开发提供借鉴。

In order to solve the problems of heavy workload and low accuracy of traditional estrus identification method in pig production,the estrous cycle of multiparous large white sows was divided into three stages:three days before oestrus(PE),three days after oestrus(E)and three days after oestrus(AE).Saliva protein was extracted from oral saliva of multiparous sows.Lable-free quantitative proteomics technique was used to detect the salivary proteome of the three stages by mass spectrometry,and the qualitative and quantitative algorithm maxquant was used for proteomics analysis.The results showed that:1)A total of 405 proteins were identified in the three stages,including149 co-existing proteins and 160 specific proteins,and 45 differential proteins were significantly screened(P<0.05).There were 54,25 and 81 specific proteins in PE,E and AE,respectively;2)GO and KEGG enrichment analysis showed that they were involved in the major metabolic pathways,including glucagon signaling pathway,pyruvate metabolism,endopeptidase inhibitor activity and protein hydrolysis regulation(P<0.05);3)Further interaction analysis showed that P79263,F1 SFI7,Q29549,F1 SS24,Q9 GMA6 and Q2 EN74 proteins had interaction relationship,among which F1 SS24、Q9 GMA6 and P79263 were PE phase proteins,F1 SFI7、Q29549 and Q2 EN74 were E-phase proteins.In conclusion,through the analysis of salivary proteome in different stages of estrus,multiple target differential/specific proteins were screened,which provided a reference for revealing the changes of salivary protein content in sow estrus cycle,and the development of estrus identification Kit/strip in the next step.