兔眼组织的二维凝胶电泳分析

Analysis of rabbit eye tissues using two-dimensional gel electrophoresis

目的 应用二维凝胶电泳对不同兔眼组织的蛋白进行初步分析 ,探索开展眼蛋白质组学的方法。方法 分离正常兔眼的不同组织 ,经样本初处理后 ,进行二维凝胶电泳和凝胶银染色 ,然后分析电泳图谱 ,比较各组织蛋白的分离效果。结果 成功建立对各眼组织进行样本处理、二维凝胶电泳的基本方法和条件。得到兔结膜、角膜内皮、虹膜、晶状体前囊膜、视网膜的二维凝胶电泳图谱 ,各组织蛋白在小胶上均能显示 4 0 0个以上的斑点。晶状体中央和周边上皮细胞的凝胶图谱呈现出数十个斑点的差异。结论 二维凝胶电泳是分析眼部组织蛋白质的一种好方法 ,不同眼部组织的蛋白在组成、含量、分布等方面的差异可通过二维凝胶电泳图谱清晰显示出来。

Objective To analyze proteins isolated from different tissues of rabbit eye using two-dimensional gel electrophoresis(2-DE). Methods The total soluble proteins were separated from different parts of rabbit eye. After sample preparation,2-DE was performed with mini gel.Followed with sil ver staining and image analyzing of 2-DE gel,the distributions of the protein spots from different tissues were displayed on the whole.Results Systemic processing and essential conditions of sample preparation and 2 -DE for various ocular tissues were established successfully.The 2-DE maps o f the conjunctiva,the corneal endothelium,the iris,the lens epithelium and the retina were obtai ne d.There were more than 400 spots with good distribution on the mini gel for eac h tissue.Decades of different spots were displayed between the central and peri pheral lens epithelial cells.Conclusion 2-DE is a powerful approach for the analysis of protein of different ocular tissues.The varieties in component,quantity and distribution of the protein s can be showed obviously on 2-DE gel.