Bershadsky A D, Tint I S, Neyfakh A A, Vasiliev J M
Exp Cell Res. 1985 Jun;158(2):433-44. doi: 10.1016/0014-4827(85)90467-7.
Morphology and distribution of cell-substrate contacts and their association with microfilament bundles in normal and RSV-transformed quail fibroblasts (16Q line) were studied by indirect immunofluorescence. The focal contacts were visualized by antibody exclusion method using monoclonal antibody to 80 kD serum protein adsorbed on the substratum. Embryo quail cells formed focal contacts of two morphological types: (1) small punctate; and (2) elongated large contacts. These two variants of contacts were designated respectively as dot and dash contacts. Both of focal contacts contained vinculin and alpha-actinin. Double immunofluorescence staining with polyclonal antibody to actin and monoclonal antibody to vinculin revealed that the dot contacts, in contrast to the dash ones, were not associated with microfilament bundles. The dot contacts were localized mostly near the active cell edges, while the dash contacts were found near the retracted cell edges and also under the central parts of the cell. We suppose that dot contacts represent initial structures which then can undergo maturation transforming them into dash contacts. RSV-transformed 16Q cells had predominantly the dot contacts which were not only located at the edges but also in the more central parts of the lamella. The dash contacts were present only in the minority of 16Q cells. RSV transformation is assumed to affect not the ability of cells to form initial dot contacts, but the maturation of dot contacts into dash contacts.
通过间接免疫荧光法研究了正常和经劳斯肉瘤病毒(RSV)转化的鹌鹑成纤维细胞(16Q系)中细胞与底物接触的形态、分布及其与微丝束的关联。使用吸附在底物上的针对80kD血清蛋白的单克隆抗体,通过抗体排除法观察粘着斑。鹌鹑胚胎细胞形成两种形态类型的粘着斑:(1)小的点状;(2)细长的大粘着斑。这两种粘着斑变体分别被命名为点状和短划线状粘着斑。两种粘着斑都含有纽蛋白和α-辅肌动蛋白。用抗肌动蛋白多克隆抗体和抗纽蛋白单克隆抗体进行双重免疫荧光染色显示,与短划线状粘着斑不同,点状粘着斑与微丝束无关。点状粘着斑大多位于活跃的细胞边缘附近,而短划线状粘着斑则出现在回缩的细胞边缘附近以及细胞的中央部分下方。我们推测点状粘着斑代表初始结构,随后可经历成熟过程转化为短划线状粘着斑。经RSV转化的16Q细胞主要具有点状粘着斑,这些粘着斑不仅位于边缘,还位于片层的更中央部分。短划线状粘着斑仅存在于少数16Q细胞中。推测RSV转化影响的不是细胞形成初始点状粘着斑的能力,而是点状粘着斑向短划线状粘着斑的成熟过程。