小鼠卵巢蛋白质双向凝胶电泳蛋白谱系的初步建立及其功能分析

目的:初步建立性成熟小鼠卵巢蛋白质图谱,并对其中的蛋白质功能进行研究.方法:通过双向电泳和质谱技术分析性成熟小鼠卵巢蛋白质组,并对其中的一种蛋白质进行免疫组化研究.结果:(1)获得高重复性和高分辨率小鼠卵巢蛋白图谱.软件分析显示,3个不同卵巢样品24 cm银染蛋白凝胶上的蛋白点均有±2 000点,胶重复性好,大部分蛋白点位于相对相同的位置.(2)质谱共鉴定出52种蛋白质,按功能对其进行分类:参与细胞代谢、细胞信号传导/交流、细胞结构/运动、细胞/器官防御与抗氧化作用、调节RNA合成、调节蛋白表达及未分类.(3)通过对其中的一种蛋白(GRP78)行免疫组化分析,发现随着卵巢由卵泡早期逐渐生长到排卵前卵泡期成熟并转向黄体期,其在颗粒细胞中的表达信号增强.结论:(1)3个不同卵巢样品蛋白质双向电泳图重复性好.我们通过质谱分析共鉴定出约52种蛋白质,初步构建了小鼠卵巢双向电泳蛋白图谱,对研究卵巢病理与生理、发育过程有着重要的参考价值.(2)行免疫组化分析结果显示,在卵巢周期变化过程中,GRP78蛋白对颗粒细胞的增生、分化、形成黄体颗粒细胞起着重要的调控作用.

A two-dimensional reference map of mouse ovary proteins

Objective:To perform a preliminary proteomic analysis of mouse ovaries and to study theprotein's function in mouse ovary.Methods:The two-dimensional gel electrophoresis(2-DE)and matrixassisted laser desorption/ionization-time of flight mass spectrometry( MALDI-TOF MS)were used to ana-lyze mouse ovarian proteome.A 12.5% sodium dodecyl sulfate(SDS)reference gel was generated byimmobilized pH gradient isoelectric focusing of mouse ovary proteins in a non-linear gradient(pH 3-10).And GRP78 was selected to perform with immunohistochemistry within mouse ovaries.Results:Based onpeptide mass fingerprinting,52 proteins were identified and classified into seven functional groups:Cell/organism defense and antioxidant,cell signaling/communications proteins,cell structure/motility pro-teins,metabolism proteins,RNA synthesis processing,Protein synthesis and processing,and unclassifiedproteins.The immunoreactivity of GRP78 was detected in GCs in the follicular,and with during GCs Lu-teinizing in the menstrual cycle,the protein expression(brown)increased continually and came to ahead when ovulation happened.Conclusion:This work provides a first step toward the establishment of asystematic ovary protein database and stands as a valuable resource for molecular analyses of normal andpathologic conditions affecting mouse ovaries.