Antras J, Hilliou F, Redziniak G, Pairault J
U 282 INSERM, Hôpital Henri-Mondor, Creteil, France.
Biol Cell. 1989;66(3):247-54.
Differentiation of 3T3-F442A cells was accompanied by changes in cell morphology, decreased synthesis and assembly of actin and fibronectin. The network of microfilament stress fibers detected with NBD-phallacidin was altered during adipose conversion of 3T3-F442A cells. Parallel to this, the disappearance of fibrillar bundles of extracellular matrix fibronectin was observed by immunofluorescence staining. The pericellular fibronectin content, detected by immunoblotting, strongly diminished during the differentiation process. An altered rate of biosynthesis of both proteins was also measured by [35S]-methionine pulse-labeling and immunoprecipitation. A 4-5-fold decrease in cellular fibronectin synthesis was observed in adipocytes compared to control preadipocytes. Conversely, non-differentiating 3T3-C2 control cells did not reorganize either the cytoskeletal architecture or the extracellular matrix fibronectin in the resting state. These results suggest that the decreased rate of biosynthesis of cell-associated fibronectin is correlated with that of actin. Moreover, both events can essentially be ascribed to differentiation.
3T3-F442A细胞的分化伴随着细胞形态的变化、肌动蛋白和纤连蛋白合成与组装的减少。在用NBD-鬼笔环肽检测时,3T3-F442A细胞脂肪转化过程中微丝应力纤维网络发生改变。与此同时,通过免疫荧光染色观察到细胞外基质纤连蛋白的纤维束消失。通过免疫印迹检测,细胞周围纤连蛋白含量在分化过程中显著减少。还通过[35S]-甲硫氨酸脉冲标记和免疫沉淀法测定了这两种蛋白质生物合成速率的变化。与对照前脂肪细胞相比,脂肪细胞中细胞纤连蛋白合成减少了4-5倍。相反,未分化的3T3-C2对照细胞在静止状态下既不重组细胞骨架结构也不重组细胞外基质纤连蛋白。这些结果表明,细胞相关纤连蛋白生物合成速率的降低与肌动蛋白的降低相关。此外,这两个事件基本上都可归因于分化。