Tassin A M, Paintrand M, Berger E G, Bornens M
J Cell Biol. 1985 Aug;101(2):630-8. doi: 10.1083/jcb.101.2.630.
In vitro myogenesis involves a dramatic reorganization of the microtubular network, characterized principally by the relocalization of microtubule nucleating sites at the surface of the nuclei in myotubes, in marked contrast with the classical pericentriolar localization observed in myoblasts (Tassin, A. M., B. Maro, and M. Bornens, 1985, J. Cell Biol., 100:35-46). Since a spatial relationship between the Golgi apparatus and the centrosome is observed in most animal cells, we have decided to follow the fate of the Golgi apparatus during myogenesis by an immunocytochemical approach, using wheat germ agglutinin and an affinity-purified anti-galactosyltransferase. We show that Golgi apparatus in myotubes displays a perinuclear distribution which is strikingly different from the polarized juxtanuclear organization observed in myoblasts. As a result, the Golgi apparatus in myotubes is situated close to the microtubule organizing center (MTOC), the cis-side being situated at a fixed distance from the nuclear envelope, a situation which suggests the existence of a structural association between the Golgi apparatus and the nuclear periphery. This is supported by experiments of microtubule depolymerization by nocodazole, in which a minimal effect was observed on Golgi apparatus localization in myotubes in contrast with the dramatic scattering observed in myoblasts. In both cell types, electron microscopy reveals that microtubule disruption generates individual dictyosomes; this suggests that the connecting structures between dictyosomes are principally affected. This structural dependency of the Golgi apparatus upon microtubules is not apparently accompanied by a reverse dependency of MTOC structure or function upon Golgi apparatus activity. Golgi apparatus modification by monensin, as effective in myotubes as in myoblasts, is without apparent effect on MTOC localization or activity and on microtubule stability. The main result of our study is to show that in a cell type where the MTOC is dissociated from centrioles and where antero-posterior polarity has disappeared, the association between the Golgi apparatus and the MTOC is maintained. The significance of such a tight association is discussed.
体外肌生成涉及微管网络的显著重组,其主要特征是肌管细胞核表面微管成核位点的重新定位,这与成肌细胞中观察到的经典的中心粒周围定位形成鲜明对比(塔辛,A.M.,B.马罗,和M.博尔南斯,1985年,《细胞生物学杂志》,100:35 - 46)。由于在大多数动物细胞中观察到高尔基体与中心体之间存在空间关系,我们决定通过免疫细胞化学方法,使用麦胚凝集素和亲和纯化的抗半乳糖基转移酶,追踪肌生成过程中高尔基体的命运。我们发现,肌管中的高尔基体呈现核周分布,这与成肌细胞中观察到的极化的近核组织明显不同。因此,肌管中的高尔基体位于靠近微管组织中心(MTOC)的位置,顺面与核膜保持固定距离,这种情况表明高尔基体与核周存在结构关联。诺考达唑诱导的微管解聚实验支持了这一点,在该实验中,与成肌细胞中观察到的高尔基体显著分散不同,肌管中高尔基体的定位受影响最小。在这两种细胞类型中,电子显微镜显示微管破坏会产生单个的高尔基体囊泡;这表明高尔基体囊泡之间的连接结构受到了主要影响。高尔基体对微管的这种结构依赖性显然并未伴随着MTOC结构或功能对高尔基体活性的反向依赖性。莫能菌素对高尔基体的修饰在肌管中和在成肌细胞中一样有效,对MTOC的定位或活性以及微管稳定性没有明显影响。我们研究的主要结果是表明,在一种MTOC与中心粒分离且前后极性消失的细胞类型中,高尔基体与MTOC之间的关联得以维持。文中讨论了这种紧密关联的意义。