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通过注射针对酪氨酸化α-微管蛋白的荧光标记抗体对活体果蝇胚胎中的微管组织和功能进行研究。

An investigation of microtubule organization and functions in living Drosophila embryos by injection of a fluorescently labeled antibody against tyrosinated alpha-tubulin.

作者信息

Warn R M, Flegg L, Warn A

机构信息

School of Biological Sciences, University of East Anglia, Norwich, United Kingdom.

出版信息

J Cell Biol. 1987 Oct;105(4):1721-30. doi: 10.1083/jcb.105.4.1721.

Abstract

Rhodamine-labeled monoclonal antibodies, which react with tyrosinated alpha-tubulin (clone YL 1/2; Kilmartin, J. V., B. Wright, and C. Milstein, 1982, J. Cell Biol., 93:576-582) and label microtubules in vivo (Wehland, J., M. C. Willingham, and I. Sandoval, 1983, J. Cell Biol., 97:1467-1475) were microinjected into syncytial stage Drosophila embryos. At 1 mg/ml antibody concentration, the microtubule arrays of the surface caps became labeled by YL 1/2 but normal development was found to continue. The results are compared with the data from fixed material particularly with regard to interphase microtubules, centrosome separation, and spindle and midbody formation. At 5 mg/ml antibody concentration the microtubules took up larger quantities of antibodies and clumped around the nuclei. Nuclei with clumped microtubules lost their position in the surface layer and moved into the interior. As a result, the F-actin cap meshwork associated with such nuclei either failed to form or subsided. It is concluded that microtubule activity is required to maintain the nuclei in the surface layer and organize the F-actin meshwork of the caps.

摘要

用若丹明标记的单克隆抗体与酪氨酸化α-微管蛋白发生反应(克隆YL 1/2;基尔马丁,J. V.,B. 赖特,和C. 米尔斯坦,1982年,《细胞生物学杂志》,93:576 - 582),并能在体内标记微管(韦兰,J.,M. C. 韦林厄姆,和I. 桑多瓦尔,1983年,《细胞生物学杂志》,97:1467 - 1475),将其显微注射到合胞体阶段的果蝇胚胎中。在抗体浓度为1毫克/毫升时,表面帽的微管阵列被YL 1/2标记,但发现正常发育仍在继续。将结果与固定材料的数据进行比较,特别是关于间期微管、中心体分离以及纺锤体和中体形成方面的数据。在抗体浓度为5毫克/毫升时,微管摄取了大量抗体并在细胞核周围聚集。微管聚集的细胞核在表层失去其位置并移入内部。结果,与这些细胞核相关的F - 肌动蛋白帽网络要么未能形成,要么消退。得出的结论是,微管活性是维持细胞核在表层并组织帽的F - 肌动蛋白网络所必需的。

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