小鼠胚胎发育不同阶段肝脏差异蛋白质组分析

利用固相pH梯度双向凝胶电泳技术分离胎龄12，13，14，16，18d的胎鼠及成鼠肝脏总蛋白，考染显色．PDQuest2DE软件分析获得6个不同发育时期的蛋白质表达谱。从总体看，14，16，18d胎鼠与成鼠肝脏的蛋白质表达谱较为一致，而12d和13d的肝脏蛋白质表达谱则与它们有较明显的差异，其蛋白质表达量显著降低，许多蛋白甚至缺失。以18d的胎肝图谱为参考，14，16d与其匹配率分别为57％，61％，成鼠与其匹配率为69％。与18d胎肝图谱相比，14d表达上调3倍以上的点有27个，16d表达上调3倍以上的点有13个，成鼠表达上调3倍以上的点有15个。并应用基质辅助激光解吸电离飞行时间质谱对14个差异蛋白质进行分析鉴定，经数据库查询，其中6个蛋白点获得了有意义的结果，分别为细胞周期依赖激酶调节亚基1、凡科尼蛋白、酪氨酸蛋白激酶BLK、通用转录因子3C多肽4、TBC1家族因子13、巨噬细胞受体MARCO。研究结果表明，在小鼠胚胎发育过程中，肝脏合成蛋白的种类和数量均迅速增加，并且细胞的增殖和生长、免疫功能的发育和完善以及蛋白质与核酸的合成和运输在其胎肝发育过程中起着非常重要的作用。

DIFFERENTIAL PROTEIOME ANALYSIS OF MOUSE LIVER IN DIFFERENT EMBRYONIC DEVELOPMENTAL STAGES

Total proteins of the liver were separated from embryo mice(E 12 d,E 13 d,E 14 d,E 16 d and E 18 d) and adult mice by immobilized pH gradient-based two-dimensional gel electrophoresis(2-DE).Their 2-DE patterns were analyzed by PDQuest Software.The proteome maps of the E 14 d,E 16 d and E 18 d fetal liver and adult liver were similar.The maps of E 12 d and E 13 d fetal liver,in which the protein content was markedly decreased and a few protein spots were lost,were significantly different from those of the E 14 d,E 16 d and E 18 d fetal liver and adult liver.When E 18 d gel acted as master,the number of matched protein spots of E 14 d,E 16 d and adult gel was 57%,61% and 69%,respectively.Compared with E 18 d gel,the spots with three-fold upexpressions were 17,13 and 15,respectively,at the E 14 d,E 16 d and adult stage.Fourteen differential expression protein spots were analyzed and identified by MALD1-TOF-MS.By sequence database searching,6 protein spots were identified.These results suggest that the liver goes through noticeable change in the expression of proteins during embryo development and their patterns and amount increase.Growth and proliferation of the cells,development and consummation of immunological function,synthesis and transport of various RNAs and proteins may play a very important role in the process of mouse embryonic liver development.