小鼠胚胎干细胞(ES-M_(13))核骨架-核纤层-中间纤维结构体系的研究  

THE INTERMEDIATE FILAMENT-LAMINA-NUCLEAR MATRIX SYSTEM OF ES-M_(13) CELLS

作　　者：高健刚[1] 韩贻仁[1] 焦仁杰[2] 翟中和[2] 

机构地区：[1]山东大学生物学系 [2]北京大学生物学系

出　　处：《实验生物学报》1994年第4期463-475,共13页Acta Biologiae Experimentalis Sinica

基　　金：山东省自然科学基金;国家自然科学基金

摘　　要：本文使用细胞的选择性抽提、DGD包埋去包埋电镜制样、免疫荧光和免疫印迹技术研究了小鼠胚胎干细胞(ES-Ml_(13))的核骨架-核纤层-中间纤维(NM-L-IF)结构体系。在电镜下可以看到,ES细胞存在精细发达的核骨架结构,核骨架纤维同核纤层结构相连接,细胞质中有许多直径为10nm的中间纤维单丝。在免疫荧光分析中,使用角蛋白单克隆抗体有阳性反应,细胞质区域可以看到较强的荧光,没有极性分布现象,也没有观察到纤维状的荧光染色。ES细胞对波形蛋白和结蛋白抗体呈阴性反应,同对照组一样,只能看到非特异性的很微弱的荧光染色。在免疫印迹分析中,使用角蛋白单克隆抗体AF6检测到三条角蛋白多肽,分子量分别为65KD,62KD和52KD。Sequential extraction and DGD embedment-free section TEM techniques were used as new methods to study the intermediate filament-lamina-nuclear matrix (IF-L-NM) system of cells, Murine embryonic stem cells (ES-M13) were investigated by means of electron microscopy, immunofluorescence microscopy and Western-blot analysis. There existed a delicate nuclear matrix network in the nucleus domain of detergent-extracted ES cells. The filaments of the nuclear matrix were found in close association with the nuclear lamina. Some intermediate filaments were also observed in the cytoplasm. In immunofluorescence microscopy, a bright, slightly granular cytoplasmic fluorescence, showing no polar concentration, was revealed with keratin monoclonal antibody AF5. We could not detect any significant fibrillar staining in the ES cells by indirect immunofluorescence method. With antibodies to vimentin and desmin, the ES cells showed only a nonspecific, weak fluorescence, similar to that seen in the control. In Western-blot analysis of electrophoretically seperated po-lypeptides, three polypeptides with molecular weight of 65 KD, 62 KD and 52 KD, reactive with keratin monoclonal antibody were detected in the ES cells.

关 键 词：小鼠 胚胎干细胞 核骨架 中间纤维 核纤层 

分 类 号：Q959.837[生物学—动物学]