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鸡胚成纤维细胞热休克及恢复过程中线粒体和中间丝的伴随变化。

Concomitant changes in mitochondria and intermediate filaments during heat shock and recovery of chicken embryo fibroblasts.

作者信息

Collier N C, Sheetz M P, Schlesinger M J

机构信息

Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110.

出版信息

J Cell Biochem. 1993 Jul;52(3):297-307. doi: 10.1002/jcb.240520306.

Abstract

Utilizing video-enhanced differential interference contrast microscopy of chicken embryo fibroblasts, we observed dramatic changes in the localization and morphology of mitochondria shortly after cells were subjected to a mild heat shock. At normal temperatures mitochondria were distributed in the cell cytoplasm as elongated, tubular, and dynamic organelles but upon heat shock they moved to the perinuclear region and formed a tight ring of short swollen and--in some cases--fused vesicles. Vital dye staining of mitochondria with rhodamine 123 and indirect immunofluorescence staining with antibodies against the mitochondrial-matrix protein, HSP 60, confirmed these results. Using cells double labeled with antibodies to vimentin and the HSP 60 protein, we found that the changes in mitochondria were accompanied by perturbations of the intermediate filament network that we and others have reported previously for heat shocked cells. Microtubules remained largely unaltered by our heat shock treatment and the redistribution of intermediate filaments and mitochondria occurred even in the presence of taxol, a microtubule stabilizing drug. The effects of heat shock on mitochondria were reversed when cells were returned to normal temperatures and their recovery to their normal state coincided with return of normal intermediate filament morphology. This recovery was blocked in cells treated with actinomycin D during heat shock, a result indicating that a heat shock protein may be required for recovery. These data are consistent with previously published observations that mitochondria are associated with the intermediate filament network but they extend this interaction to a cell system responding to a physiological stress normally experienced by the intact organism.

摘要

利用视频增强型微分干涉差显微镜观察鸡胚成纤维细胞,我们发现细胞受到轻度热休克后不久,线粒体的定位和形态发生了显著变化。在正常温度下,线粒体以细长、管状且动态的细胞器形式分布于细胞质中,但热休克后它们会移至核周区域,并形成由短而肿胀且在某些情况下融合的囊泡组成的紧密环。用罗丹明123对线粒体进行活细胞染料染色以及用抗线粒体基质蛋白HSP 60的抗体进行间接免疫荧光染色,证实了这些结果。使用针对波形蛋白和HSP 60蛋白的抗体对细胞进行双重标记,我们发现线粒体的变化伴随着中间丝网络的扰动,这是我们和其他人之前报道过的热休克细胞的情况。我们的热休克处理对微管的影响不大,即使在存在微管稳定药物紫杉醇的情况下,中间丝和线粒体的重新分布仍会发生。当细胞恢复到正常温度时,热休克对线粒体的影响会逆转,并且它们恢复到正常状态与中间丝形态恢复正常同时发生。在热休克期间用放线菌素D处理的细胞中,这种恢复受到阻碍,这一结果表明恢复可能需要一种热休克蛋白。这些数据与之前发表的观察结果一致,即线粒体与中间丝网络相关,但它们将这种相互作用扩展到一个对完整生物体通常经历的生理应激作出反应的细胞系统。

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