无标记定量方法在小鼠精子发生相关蛋白质组学研究中的应用

目的: 利用无标记定量蛋白质组学技术筛选在3周龄及成年小鼠睾丸中差异表达的蛋白质?方法:分别抽提3周龄及成年小鼠睾丸蛋白,酶解后所得肽段经纳升液相色谱分离,并经LTQ Obitrap质谱鉴定,在获得蛋白鉴定信息的同时根据相应肽段离子质谱峰强度对蛋白进行相对定量?结果:在3周龄及成年小鼠睾丸中共鉴定到非冗余蛋白319种,其中30种蛋白在两者之间存在显著的表达差异,这些蛋白主要参与形态发生?收缩?胞吞?受精等细胞事件,与精子变形过程及精子功能相关?结论:无标记定量蛋白质组学技术,具有方法简便?通量大?成本低廉等优点,利用其寻找睾丸发育,精子发生/功能相关蛋白具有广阔的前景?

The application of label-free quantitative method in mouse spermatogenesis related proteome

Objective: To identify the proteins differently expressed between 3-week-old and adult mouse testes by label-free quantitative approach. Methods:Proteins were extracted respectively from 3-week-old and adult mouse testes,and digested by trypsin and subsequently analyzed by Nano LC-(liquid chromatography)-LTQ Orbitrap Mass Spectrometric Method. Using this method,the proteins were identified,while at the same time,the relative quantitation of protein expression was calculated according to peak intensity. Results:A grand total of 319 non-redundant proteins were identified,and 30 proteins among them were observed to exhibit significantly differential expression between 3-week-old and adult mouse testes(P < 0.05). These proteins primarily participate in morphogenesis,contraction,endocytosis and fertilization. These processes were tightly related with the spermiogenesis and sperm function. Conclusion:The label-free quatitative proteomic technique provided simple,high- throughput and low-cost measurement of protein expression levels. As such,it has a broad prospect in identifying related proteins involved in testis development,spermatogenesis and sperm function.