Expression of intermediate filaments in cultured cells.
Expression of intermediate filaments in cultured cells.
复制标题
培养细胞中中间丝的表达。
作者：
I. Virtanen;V. Lehto;E. Lehtonen;T. Vartio;S. Stenman;P. Kurki;O. Wager;J. Small;D. Dahl;R. A. Badley
采用间接免疫荧光（IFL）技术研究了原代培养细胞和培养细胞系中不同类型中间丝的发生。使用的抗体是针对波形蛋白型中间丝（成纤维细胞58 X IOmol. wt亚基蛋白），和针对波形蛋白、结蛋白的IgG类实验兔抗体（肌肉55 X IOmol. wt亚基蛋白）、胶质细胞酸性蛋白（GFA）、68 X IOmol. WT神经丝多肽和人角蛋白多肽。培养的成纤维细胞从不同的物种，主动脉和静脉内皮细胞，显示了纤维状的细胞质荧光时，对波形蛋白的抗体染色。另一方面，在鸡胚胎成纤维细胞的培养物中，即使经过几次传代培养，也可以看到显示明亮的结蛋白特异性荧光但缺乏波形蛋白特异性染色的细胞。聚丙烯酰胺凝胶电泳也证实了这些培养物中存在结蛋白和波形蛋白多肽。在鸡成纤维细胞培养物中，2种类型的中间丝在单个细胞中不同时表达，而在幼仓鼠肾（BHK-21）细胞和人横纹肌肉瘤细胞中，可以看到抗波形蛋白和抗结蛋白抗体的可变共染色。相反，培养的人纤维肉瘤细胞和猿猴病毒40转化的人成纤维细胞仅显示波形蛋白特异性纤维荧光。来自小鼠胚胎脊髓的胶质细胞似乎在原代培养中仅表达含GFA的中间丝，而继代培养的小鼠胶质细胞和培养的胶质瘤细胞系也显示波形蛋白特异性染色。另一方面，原代培养物中的神经元样细胞只能用68 × 10 mol的抗体染色。wt神经丝多肽。有趣的是，小鼠神经母细胞瘤（Ci 300）细胞克隆含有
The occurrence of different types of intermediate filaments in primary cultures of cells and in cultured cell lines was studied by the indirect immunofluorescence (IFL) technique. The antibodies used were spontaneous monoclonal human antibodies of immunoglobulin M (IgM) class against vimentin-type intermediate filaments (fibroblast 58 x 10 mol. wt subunit protein), and experimental rabbit antibodies of IgG class against vimentin, desmin (muscle 55 x io mol. wt subunit protein), glial fibrillary acidic protein (GFA), 68 x io mol. wt neurofilament polypeptide and human keratin polypeptides. Cultured fibroblasts from different species, and both aortic and venous endothelial cells, showed a fibrillar cytoplasmic fluorescence when stained with antibodies against vimentin. On the other hand, in cultures of chicken embryonal fibroblasts, cells showing a bright desminspecific fluorescence, but lacking vimentin-specific staining, were seen even after several subcultivations. The presence of both desmin and vimentin polypeptides in these cultures was also confirmed by polyacrylamide gel electrophoresis. In chicken fibroblast cultures the 2 types of intermediate filaments were not expressed simultaneously in individual cells, whereas in baby hamster kidney (BHK-21) cells and human rhabdomyosarcoma cells a variable co-staining with anti-vimentin and anti-desmin antibodies could be seen. In contrast, cultured human fibrosarcoma cells and simian virus 40-transformed human fibroblasts showed only vimentinspecific fibrillar fluorescence. Glial cells from mouse embryonic spinal cord appeared to express only GFA-containing intermediate filaments in a primary culture, whereas both subcultured mouse glial cells and a cultured glioma cell-line also showed vimentin-specific staining. On the other hand, neuronelike cells in the primary cultures could only be stained with the antibodies to 68 x io mol. wt neurofilament polypeptide. Interestingly, clones ofmouseneuroblastoma(Ci 300) cells contained