免疫胶体金标记显示波形纤维与核孔复合体的结合

中间纤维与细胞核的关系是一个亟待解决的重要问题。本文采用火鸡红细胞作为研究材料,首先用细胞分级抽提结合免疫印迹反应显示火鸡红细胞中间纤维蛋白为波形纤维蛋白。然后,我们采用细胞分级抽提结合包埋前免疫胶体金标记的方法显示胞质中间纤维被抗波形纤维蛋白抗体-蛋白A-胶体金特异标记。同时,我们显示结合于核孔复合体上的胞质纤维被抗波形纤维蛋白抗体-蛋白A-胶体金所特异标记。本文结果表明,结合于核孔复合体上的胞质纤维是波形纤维,并且提示波形纤维可能通过与Nup 180的结合附着在核孔复合体上,为进一步探讨中间纤维与核孔运输的关系提供了初步实验证据。

IMMUNOGOLD LABELING ELECTRON MICROSCOPY SHOWING VIMENTIN FILAMENT ANCHORED ON NUCLEAR PORE COMPLEX

The relationship between intermidia-te filament and nucleus is an important question to be solved. By combining sequential cell fractionation with immuno-blotting, we showed that the intermediate filament protein in turkey erythrocyte is vimentin. Using pre-embedment immuno-gold labeling techniques together with sequential cell fractionation, we showed that cytoplasmic intermediate filaments in turkey erythrocyte are labeled specifically by rabbit anti-vimentin antibody- protein A-gold. Moreover, we showed that the cytoplasmic filaments anchored on nuclear pore complex were also labeled by rabbit anti-vimentin antibody-protein A-gold specifically. Our results demonstrated that cytoplasmic filaments anchored on nuclear pore complex was vimentin filament. The experiments indicated that vimentin filament may be anchored on nuclear pore complex by binding with Nup 180 and intermediate filament may be involved in nuclear transportation.