Chaly N, Bladon T, Setterfield G, Little J E, Kaplan J G, Brown D L
J Cell Biol. 1984 Aug;99(2):661-71. doi: 10.1083/jcb.99.2.661.
We examined the distribution of nonlamin nuclear matrix antigens during the mitotic cell cycle in mouse 3T3 fibroblasts. Four monoclonal antibodies produced against isolated nuclear matrices were used to characterize antigens by the immunoblotting of isolated nuclear matrix preparations, and were used to localize the antigens by indirect immunofluorescence. For comparison, lamins and histones were localized using human autoimmune antibodies. At interphase, the monoclonal antibodies recognized non-nucleolar and nonheterochromatin nuclear components. Antibody P1 stained the nuclear periphery homogeneously, with some small invaginations toward the interior of the nucleus. Antibody I1 detected an antigen distributed as fine granules throughout the nuclear interior. Monoclonals PI1 and PI2 stained both the nuclear periphery and interior, with some characteristic differences. During mitosis, P1 and I1 were chromosome-associated, whereas PI1 and PI2 dispersed in the cytoplasm. Antibody P1 heavily stained the periphery of the chromosome mass, and we suggest that the antigen may play a role in maintaining interphase and mitotic chromosome order. With antibody I1, bright granules were distributed along the chromosomes and there was also some diffuse internal staining. The antigen to I1 may be involved in chromatin/chromosome higher-order organization throughout the cell cycle. Antibodies PI1 and PI2 were redistributed independently during prophase, and dispersed into the cytoplasm during prometaphase. Antibody PI2 also detected antigen associated with the spindle poles.
我们研究了小鼠3T3成纤维细胞有丝分裂细胞周期中非核纤层核基质抗原的分布。利用针对分离的核基质产生的四种单克隆抗体,通过对分离的核基质制剂进行免疫印迹来鉴定抗原,并通过间接免疫荧光对抗原进行定位。作为对照,使用人类自身免疫抗体对核纤层蛋白和组蛋白进行定位。在间期,单克隆抗体识别非核仁及非异染色质的核成分。抗体P1均匀地染色核周边,向核内部有一些小的内陷。抗体I1检测到一种呈细颗粒状分布于整个核内的抗原。单克隆抗体PI1和PI2同时染色核周边和内部,存在一些特征性差异。在有丝分裂期间,P1和I1与染色体相关,而PI1和PI2分散在细胞质中。抗体P1强烈染色染色体团块的周边,我们认为该抗原可能在维持间期和有丝分裂期染色体秩序中发挥作用。使用抗体I1时,明亮的颗粒沿染色体分布,并且也有一些弥散的内部染色。I1的抗原可能在整个细胞周期中参与染色质/染色体的高级组织。抗体PI1和PI2在前期独立重新分布,并在中期前分散到细胞质中。抗体PI2还检测到与纺锤体极相关的抗原。