INTERRELATIONSHIPS OF ENDOPLASMIC-RETICULUM, MITOCHONDRIA, INTERMEDIATE FILAMENTS, AND MICROTUBULES - A QUADRUPLE FLUORESCENCE LABELING STUDY
INTERRELATIONSHIPS OF ENDOPLASMIC-RETICULUM, MITOCHONDRIA, INTERMEDIATE FILAMENTS, AND MICROTUBULES - A QUADRUPLE FLUORESCENCE LABELING STUDY
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DOI：
10.1139/o92-163
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发表时间：
1992-10-01
期刊：
影响因子：
--
通讯作者：
GUPTA, RS
中科院分区：
文献类型：
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作者：
SOLTYS, BJ;GUPTA, RS
为了研究内质网、线粒体、中间细丝和微管的相互关系，我们开发了一种四重荧光标记程序来可视化同一细胞中的所有四种结构。我们应用这种方法来研究对照细胞和用微管药物长春花碱或紫杉醇处理的细胞的细胞组织。内质网通过用3,3'-二己基氧碳蓝碘化染料染色戊二醛固定细胞可见。洗涤剂渗透后，进行三重免疫荧光特异性观察线粒体，波形蛋白中间丝和微管。人类成纤维细胞中的线粒体被发现是高度细长的管状结构（长度超过50微米），在许多情况下，它们明显融合在一起。虽然内质网也独立存在，但线粒体总是与内质网相关。中间纤维的分布不能完全解释内质网或线粒体的分布。然而，微管总是与这些细胞器共分布。长春碱处理后的细胞微管解聚导致内质网和线粒体共聚集，中间丝断裂。与中间丝相比，细胞器的空间分布不相同，说明细胞器与中间丝的附着不是细胞器聚集的原因。另一方面，线粒体与内质网的关联得以保留，表明无论内质网形式或微管如何，这种关联都是稳定的。在紫杉醇处理的细胞中，内质网、线粒体和中间丝都与紫杉醇稳定的微管束相关。
To study the interrelationships of endoplasmic reticulum, mitochondria, intermediate filaments, and microtubules, we have developed a quadruple fluorescence labeling procedure to visualize all four structures in the same cell. We applied this approach to study cellular organization in control cells and in cells treated with the microtubule drugs vinblastine or taxol. Endoplasmic reticulum was visualized by staining glutaraldehyde-fixed cells with the dye 3,3'-dihexyloxacarbocyanine iodide. After detergent permeabilization, triple immunofluorescence was carried out to specifically visualize mitochondria, vimentin intermediate filaments, and microtubules. Mitochondria in human fibroblasts were found to be highly elongated tubular structures (lengths up to greater than 50 mum), which in many cases were apparently fused to each other. Mitochondria were always observed to be associated with endoplasmic reticulum, although endoplasmic reticulum also existed independently. Intermediate filament distribution could not completely account for endoplasmic reticulum or mitochondrial distributions. Microtubules, however, always codistributed with these organelles. Microtubule depolymerization in vinblastine treated cells resulted in coaggregation of endoplasmic reticulum and mitochondria, and in the collapse of intermediate filaments. The spatial distributions of organelles compared with intermediate filaments were not identical, indicating that attachment of organelles to intermediate filaments was not responsible for organelle aggregation. Mitochondrial associations with endoplasmic reticulum, on the other hand, were retained, indicating this association was stable regardless of endoplasmic reticulum form or microtubules. In taxol-treated cells, endoplasmic reticulum, mitochondria, and intermediate filaments were all associated with taxol- stabilized microtubule bundles.