Mounier N, Perriard J C, Gabbiani G, Chaponnier C
Centre de Génétique Moléculaire et Cellulaire, Université Lyon 1, Villeurbanne, France.
J Cell Sci. 1997 Apr;110 ( Pt 7):839-46. doi: 10.1242/jcs.110.7.839.
We have analyzed by immunolabeling the fate of exogenous epitope-tagged actin isoforms introduced into cultured smooth muscle and non-muscle (i.e. endothelial and epithelial) cells by transfecting the corresponding cDNAs in transient expression assays. Exogenous muscle actins did not produce obvious shape changes in transfected cells. In smooth muscle cells, transfected striated and smooth muscle actins were preferentially recruited into stress fibers. In non-muscle cells, exogenous striated muscle actins were rarely incorporated into stress fibers but remained scattered within the cytoplasm and frequently appeared organized in long crystal-like inclusions. Transfected smooth muscle actins were incorporated into stress fibers of epithelial cells but not of endothelial cells. Exogenous non-muscle actins induced alterations of cell architecture and shape. All cell types transfected by non-muscle actin cDNAs showed an irregular shape and a poorly developed network of stress fibers. beta- and gamma-cytoplasmic actins transfected into muscle and non-muscle cells were dispersed throughout the cytoplasm, often accumulated at the cell periphery and rarely incorporated into stress fibers. These results show that isoactins are differently sorted: not only muscle and non-muscle actins are differentially distributed within the cell but also, according to the cell type, striated and smooth muscle actins can be discriminated for. Our observations support the assumption of isoactin functional diversity.
我们通过免疫标记分析了在瞬时表达试验中通过转染相应cDNA将外源表位标记的肌动蛋白异构体引入培养的平滑肌和非肌肉(即内皮和上皮)细胞后的命运。外源肌肉肌动蛋白在转染细胞中未产生明显的形态变化。在平滑肌细胞中,转染的横纹肌和平滑肌肌动蛋白优先被募集到应力纤维中。在非肌肉细胞中,外源横纹肌肌动蛋白很少掺入应力纤维,而是散在于细胞质中,并经常呈长晶体状内含物排列。转染的平滑肌肌动蛋白被掺入上皮细胞的应力纤维中,但未掺入内皮细胞的应力纤维中。外源非肌肉肌动蛋白诱导细胞结构和形态的改变。所有用非肌肉肌动蛋白cDNA转染的细胞类型均呈现不规则形状且应力纤维网络发育不良。转染到肌肉和非肌肉细胞中的β-和γ-细胞质肌动蛋白分散在整个细胞质中,经常聚集在细胞周边,很少掺入应力纤维中。这些结果表明不同的肌动蛋白异构体被不同地分选:不仅肌肉和非肌肉肌动蛋白在细胞内的分布不同,而且根据细胞类型,横纹肌和平滑肌肌动蛋白也可以被区分。我们的观察结果支持肌动蛋白异构体功能多样性的假设。