Verheijen R, Kuijpers H J, van Driel R, Beck J L, van Dierendonck J H, Brakenhoff G J, Ramaekers F C
Department of Pathology, University Hospital Nijmegen, The Netherlands.
J Cell Sci. 1989 Apr;92 ( Pt 4):531-40. doi: 10.1242/jcs.92.4.531.
In interphase cells the proliferation-associated antigen recognized by monoclonal antibody Ki-67 is almost exclusively located in the nucleoli. When cells at several stages of mitosis were examined for the localization of the Ki-67 antigen, a striking redistribution could be observed. During prophase the distinct nucleolar Ki-67 fluorescence changed to a bright irregular meshwork throughout the nucleoplasm. At metaphase the antigen appeared to be distributed in a reticulate structure surrounding the condensed chromosomes, while at late telophase a punctated staining of the entire nucleoplasm was observed, which preceded the typical nucleolar localization pattern in each of the two daughter cells. Immunolabelling with Ki-67 of metaphase chromosome spreads revealed a circumferential staining of the individual chromosomes. The Ki-67 antigen is preserved in nuclear matrix preparations obtained after in situ fractionation of interphase cells. When mitotic cells were exposed to such treatments, the obtained fluorescence data suggested that the antigen may be part of the chromosome scaffold. Quantification of the Ki-67 fluorescence signal using flow cytometry revealed the highest staining intensities in mitotic cells. Furthermore, it was shown that nutritionally deprived cells became negative for Ki-67.
在间期细胞中，单克隆抗体Ki-67识别的增殖相关抗原几乎仅位于核仁中。当检查处于有丝分裂几个阶段的细胞中Ki-67抗原的定位时，可以观察到明显的重新分布。在前期，明显的核仁Ki-67荧光变为整个核质中的明亮不规则网络。在中期，抗原似乎分布在围绕浓缩染色体的网状结构中，而在末期后期，观察到整个核质有斑点状染色，这在两个子细胞中典型的核仁定位模式之前出现。用Ki-67对中期染色体铺展进行免疫标记显示单个染色体有周边染色。Ki-67抗原保留在间期细胞原位分级分离后获得的核基质制剂中。当有丝分裂细胞接受此类处理时，获得的荧光数据表明该抗原可能是染色体支架的一部分。使用流式细胞术对Ki-67荧光信号进行定量分析显示有丝分裂细胞中的染色强度最高。此外，研究表明营养缺乏的细胞对Ki-67呈阴性。
