Turner B M, Ruane M
Eur J Cell Biol. 1985 Jan;36(1):48-57.
The organisation of cytokeratin filaments in mitotic HeLa cells has been analysed by immunofluorescence microscopy using a monoclonal antibody which recognises proteins with apparent subunit molecular weights of 52 kDa and 57 kDa and which binds exclusively to cytokeratin-type filaments. Mitotic cells were prepared for microscopic analysis by hypotonic swelling, centrifugation onto glass slides, brief pre-extraction with 0.1% Triton X-100 and fixation in 80% ethanol. This procedure gave particularly good resolution of intermediate filaments and preservation of chromosome morphology. In prometaphase-metaphase cells the antigen was present in an anastomosing filament network which completely or partially enclosed the chromosomes, in filament fragments and in cytoplasmic aggregates. The epichromosomal filament network was absent from cells in anaphase or later stages of mitosis. In these cells non-filamentous antigen was often located in a narrow band defining the periphery of individual chromosomes and in variable numbers of cytoplasmic filaments or fragments. The results suggest that extensive disaggregation and reformation of cytokeratin filaments occurs during mitosis and that disaggregated cytokeratin proteins are frequently located adjacent to mitotic chromosomes.
利用一种单克隆抗体，通过免疫荧光显微镜分析了有丝分裂期海拉细胞中细胞角蛋白丝的组织情况。该单克隆抗体可识别表观亚基分子量为52 kDa和57 kDa的蛋白质，且仅与细胞角蛋白类型的丝结合。通过低渗肿胀、离心到载玻片上、用0.1% Triton X-100进行短暂预抽提以及在80%乙醇中固定等步骤，制备用于显微镜分析的有丝分裂细胞。该方法能特别清晰地显示中间丝，并保留染色体形态。在前中期至中期细胞中，抗原存在于一个相互吻合的丝网络中，该网络完全或部分包围染色体，也存在于丝片段和细胞质聚集体中。后期或有丝分裂后期的细胞中不存在染色体表面丝网络。在这些细胞中，非丝状抗原通常位于界定单个染色体周边的窄带中，以及数量不等的细胞质丝或片段中。结果表明，细胞角蛋白丝在有丝分裂期间会发生广泛的解聚和重新形成，且解聚的细胞角蛋白蛋白经常位于有丝分裂染色体附近。
