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单克隆抗体揭示的纤毛上皮细胞顶端极的肌球蛋白

Myosin at the apical pole of ciliated epithelial cells as revealed by a monoclonal antibody.

作者信息

Klotz C, Bordes N, Laine M C, Sandoz D, Bornens M

出版信息

J Cell Biol. 1986 Aug;103(2):613-9. doi: 10.1083/jcb.103.2.613.

Abstract

A monoclonal antibody (CC-212), obtained in a fusion experiment in which basal bodies from quail oviduct were used as immunogen, has been shown to label the apical pole of ciliated cells and to react with a 200-kD protein. This monoclonal antibody was demonstrated to be an anti-myosin from smooth muscle or from nonmuscular cells using the following criteria: On Western blots it reacted with the myosin heavy chains from gizzard and platelet extracts and from cultured cell line extracts, but did not react with striated muscle myosin heavy chains. By immunofluorescence it decorated the stress fibers of well-spread cells with a characteristic striated pattern, while it did not react with myotubes containing organized myofibrils. On native ciliated cells as well as on Triton-extracted ciliated cortices from quail oviduct, this monoclonal antibody decorated the apical pole with a stronger labeling of the periphery of the apical area. Ultrastructural localization was attempted using the immunogold technique on the same preparation. Myosin was associated with a filamentous material present between striated rootlets and the proximal extremities of the basal bodies. No labeling of the basal body itself or of axoneme was observed.

摘要

在一项融合实验中获得了一种单克隆抗体(CC - 212),该实验使用鹌鹑输卵管的基体作为免疫原,已证明它可标记纤毛细胞的顶端极,并与一种200-kD的蛋白质发生反应。使用以下标准证明该单克隆抗体是一种来自平滑肌或非肌肉细胞的抗肌球蛋白:在蛋白质免疫印迹法中,它与来自砂囊、血小板提取物以及培养细胞系提取物中的肌球蛋白重链发生反应,但不与横纹肌肌球蛋白重链发生反应。通过免疫荧光法,它以特征性的条纹模式装饰充分铺展细胞的应力纤维,而不与含有有组织肌原纤维的肌管发生反应。在鹌鹑输卵管的天然纤毛细胞以及经Triton处理提取的纤毛皮质上,该单克隆抗体标记顶端极,顶端区域周边的标记更强。尝试使用免疫金技术在同一制剂上进行超微结构定位。肌球蛋白与存在于横纹小根和基体近端之间的丝状物质相关。未观察到基体本身或轴丝的标记。

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本文引用的文献

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Preparation of monoclonal antibodies: strategies and procedures.单克隆抗体的制备:策略与程序
Methods Enzymol. 1981;73(Pt B):3-46. doi: 10.1016/0076-6879(81)73054-4.

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