Laurila P, Virtanen I, Lehto V P, Vartio T, Stenman S
J Cell Biol. 1982 Aug;94(2):308-15. doi: 10.1083/jcb.94.2.308.
The expression of intermediate filaments of the keratin- and the vimentin-type was studied in heterokaryons of human fibroblasts and amnion epithelial cells by immunofluorescence microscopy. Fibroblasts and their homokaryons showed a fibrillar, vimentin-specific fluorescence throughout the cytoplasm but were negative when stained for keratin. Amnion epithelial cells and their homokaryons, on the other hand, showed a keratin-specific fibrillar staining, and only some of them contained also detectable vimentin. When suspended epithelial cells were fused with adherent fibroblasts, keratin fibrils spread within 3 h into the fibroblasts, intermixing with the vimentin fibrils. 1-3 d after fusion, both vimentin and keratin filaments were expressed as typical fibrillar cytoplasmic arrays, and the distribution of keratin in heterokaryons resembled closely that of vimentin. A typical cell-to-cell arrangement of keratin fibrils, seen in cultures of amnion epithelial cells, could also be found between heterokaryons. Treatment of the cultures with vinblastine sulphate induced coiling of the vimentin filaments in both homo- and heterokaryons, whereas the keratin organization was only slightly affected. Our results show that both vimentin and keratin filaments are incorporated into the cytoskeleton of heterokaryons formed between fibroblasts and epithelial cells, and that they behave in the same way as in their parental cells. Both epithelial and fibroblastic characteristics thus appear to the coexpressed in such heterokaryons.
通过免疫荧光显微镜技术,研究了人成纤维细胞与羊膜上皮细胞异核体中角蛋白型和波形蛋白型中间丝的表达情况。成纤维细胞及其同核体在整个细胞质中呈现出纤维状的、波形蛋白特异性荧光,但用角蛋白染色时呈阴性。另一方面,羊膜上皮细胞及其同核体显示出角蛋白特异性的纤维状染色,并且只有其中一些细胞也含有可检测到的波形蛋白。当悬浮的上皮细胞与贴壁的成纤维细胞融合时,角蛋白纤维在3小时内扩散到成纤维细胞中,与波形蛋白纤维混合。融合后1 - 3天,波形蛋白和角蛋白丝均表达为典型的纤维状细胞质阵列,异核体中角蛋白的分布与波形蛋白的分布非常相似。在异核体之间也可以发现羊膜上皮细胞培养物中所见的角蛋白纤维典型的细胞间排列。用硫酸长春碱处理培养物会导致同核体和异核体中波形蛋白丝卷曲,而角蛋白的组织仅受到轻微影响。我们的结果表明,波形蛋白丝和角蛋白丝都被纳入成纤维细胞和上皮细胞之间形成的异核体的细胞骨架中,并且它们在异核体中的行为与其亲代细胞中的行为相同。因此,上皮和成纤维细胞的特征似乎在这种异核体中共同表达。