Heterogeneity among microtubules of the cytoplasmic microtubule complex detected by a monoclonal antibody to alpha tubulin.
Heterogeneity among microtubules of the cytoplasmic microtubule complex detected by a monoclonal antibody to alpha tubulin.
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DOI：
10.1083/jcb.98.3.1017
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发表时间：
1984-03
期刊：
影响因子：
--
通讯作者：
Carney DH
中科院分区：
文献类型：
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作者：
Thompson WC;Asai DJ;Carney DH
通过间接免疫荧光法检测了三种微管蛋白特异性单克隆抗体对小鼠和人成纤维细胞胞质微管的染色能力。我们使用双标记免疫荧光比较这些抗体的染色模式与用多克隆兔抗微管蛋白试剂染色的相同细胞中的总微管复合物。两个单克隆抗微管蛋白抗体结合所有的细胞质微管，但抗体1-6。1结合的细胞质微管的一个子集内的个别固定的细胞。在各种固定条件和染色方案下，在洗涤剂提取的细胞骨架以及在整个固定的细胞中观察到差异染色模式。至少有一个生理上定义的细胞质微管的子集，那些保留在细胞预处理1小时，5 μ M秋水仙胺，似乎完全由抗体1-6。1例阳性微管。无论是在冷处理的细胞中，还是在暴露于低渗介质的细胞中，微管都没有出现同样的情况。单个固定细胞内微管之间的抗原差异的演示和这种抗原差异与至少一个“生理学”定义的子集的明显相关性表明，存在个别微管在体内的差异组装或组装后修饰的机制，这可能赋予它们不同的物理或功能特性。
Three monoclonal antibodies specific for tubulin were tested by indirect immunofluorescence for their ability to stain cytoplasmic microtubules of mouse and human fibroblastic cells. We used double label immunofluorescence to compare the staining patterns of these antibodies with the total microtubule complex in the same cells that were stained with a polyclonal rabbit antitubulin reagent. Two of the monoclonal antitubulin antibodies bound to all of the cytoplasmic microtubules but Ab 1-6. 1 bound only a subset of cytoplasmic microtubules within individual fixed cells. Differential staining patterns were observed under various fixation conditions and staining protocols, in detergent-extracted cytoskeletons as well as in whole fixed cells. At least one physiologically defined subset of cytoplasmic microtubules, those remaining in cells pretreated for 1 h with 5 microM colcemid, appeared to consist entirely of Ab 1-6. 1 positive microtubules. The same was not true of the microtubules that remained in either cold-treated cells or in cells that had been exposed to hypotonic medium. The demonstration of antigenic differences among microtubules within single fixed cells and the apparent correlation of this antigenic difference with at least one "physiologically" defined subset suggests that mechanisms exist for the differential assembly or postassembly modification of individual microtubules in vivo, which may endow them with different physical or functional properties.