DISRUPTION OF THE INVIVO DISTRIBUTION OF THE INTERMEDIATE FILAMENTS IN FIBROBLASTS THROUGH THE MICRO-INJECTION OF A SPECIFIC MONOCLONAL-ANTIBODY
DISRUPTION OF THE INVIVO DISTRIBUTION OF THE INTERMEDIATE FILAMENTS IN FIBROBLASTS THROUGH THE MICRO-INJECTION OF A SPECIFIC MONOCLONAL-ANTIBODY
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DOI：
10.1016/0092-8674(81)90514-6
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发表时间：
1981-01-01
期刊：
影响因子：
64.5
通讯作者：
FERAMISCO, JR
中科院分区：
文献类型：
--
作者：
LIN, JJC;FERAMISCO, JR
通过显微注射将识别培养细胞的中间丝系统的次要组分的单克隆抗体（JLB 1和JLB 7）引入活的成纤维细胞中。注射JLB 7抗体几分钟后，发现几乎所有细胞的中间丝都在细胞核附近或周围聚集成紧密的束。相比之下，注射JLB 1抗体引起很少或不引起中间丝的聚集。电子显微镜显示，注射JLB 7抗体后形成的核周束，每个都由10个或更多个明显交联在一起的中间丝组成。双标记免疫荧光显微镜显示，几乎所有的波形蛋白含有中间丝在JLB 7抗体注射细胞被重新分配到核周区域，并保持在那里至少24 hr. The分布的肌动蛋白微丝和微管似乎不受干扰后显微注射。在注射JLB 7抗体的细胞中，细胞形态或行为没有明显的变化;细胞显示出扁平的外观，显示出极性，能够起皱和起泡，甚至似乎显示出胞内囊泡、颗粒和线粒体的正常跳跃运动，表明中间丝不参与这些活动。显微注射识别和改变细胞成分的高度特异性单克隆抗体提供了确定细胞内元件的体内功能的另一种方法。
Monoclonal antibodies(JLBl and JLB7) that recognize minor components of the intermediate filament system of cultured cells were introduced into living fibroblasts by microinjection. Several minutes after injection of the JLB7 antibody virtually all of the intermediate filaments of the cells were found to be aggregated into tight bundles near or around the nucleus. In contrast, injection of the JLBl antibody caused little or no aggregation of the intermediate filaments. Electron microscopy showed that the perinuclear bundles that formed after injection of the JLB7 antibody each consisted of ten or more intermediate filaments apparently crosslinked together. Double-label immunofluorescence microscopy showed that virtually all of the vimentin-containing intermediate filaments in the JLB7 antibody-injected cells were redistributed to the perinuclear region and remained there for at least 24 hr. The distributions of actin microfilaments and microtubules were seemingly undisturbed following microinjection. No obvious changes in cell morphology or behavior were apparent in the cells injected with JLB7 antibody; the cells displayed a flat appearance, showed a polarity, were able to ruffle and bleb and even appeared to show the normal saltatory movements of intracellular vesicles, granules and mitochondria, suggesting that intermediate filaments are not involved in these activities. The microinjection of highly specific monoclonal antibodies that recognize and alter components of the cell provides an additional approach to determine the in vivo functions of intracellular elements.