Yates J R, Izzard C S
J Cell Sci. 1981 Dec;52:183-96. doi: 10.1242/jcs.52.1.183.
The cell-to-substrate contacts of the adhesion-defective mutant, AD6, have been examined by interference reflexion microscopy and compared with those of the wild-type Balb/c3T3 cell. The 2 cell-types differed in their ability to produce focal contacts with the substrate (10-15 nm separation distance). Only 10% of AD6 versus 92% of Balb/c3T3 cells formed these contacts. When present in AD6 the focal contacts were smaller and fewer in number per cell than in Balb/c3T3. Close contact with the substrate (approx. 30 nm separation distance) was formed by both cell types. The absence of the stronger focal contacts accounts for the reduced substrate adhesion of the mutant cells and for other phenotypic characteristics such as reduced spreading, rounded shape, altered pattern of movement, and absence of stress fibres. The reduced adhesion of the mutant has been attributed to a change in the pattern of glycoproteins exposed at the cell surface, which results from a defect in glycosylation. This suggests that normal glycosylation and correct exposure of one or more cell surface components are required for the formation of a specific substrate adhesion, the focal contact, and offers the possibility of identifying individual surface components involved in formation of this adhesion. Attention is drawn to zero-order minima originating from cell thickness and not cell-to-substrate separation distance in the interference reflexion image.
通过干涉反射显微镜对黏附缺陷型突变体AD6的细胞与底物接触情况进行了检测,并与野生型Balb/c3T3细胞的接触情况进行了比较。这两种细胞类型在与底物形成粘着斑(分离距离为10 - 15纳米)的能力上存在差异。AD6细胞中只有10%能形成这些粘着斑,而Balb/c3T3细胞的这一比例为92%。在AD6细胞中,若存在粘着斑,其尺寸比Balb/c3T3细胞中的更小,且每个细胞中的数量也更少。两种细胞类型都能与底物形成紧密接触(分离距离约为30纳米)。突变细胞缺乏更强的粘着斑,这导致了其对底物的黏附力降低,以及其他表型特征,如铺展减少、形状变圆、运动模式改变和应力纤维缺失。突变体黏附力降低被归因于细胞表面暴露的糖蛋白模式发生了变化,这是由糖基化缺陷导致的。这表明正常的糖基化以及一种或多种细胞表面成分的正确暴露是形成特定底物黏附即粘着斑所必需的,并且提供了识别参与这种黏附形成的单个表面成分的可能性。需要注意的是,干涉反射图像中源于细胞厚度而非细胞与底物分离距离的零级极小值。