O'Neill C, Jordan P, Riddle P, Ireland G
Imperial Cancer Research Fund, Lincoln's Inn Fields, London, UK.
J Cell Sci. 1990 Apr;95 ( Pt 4):577-86. doi: 10.1242/jcs.95.4.577.
The stimulus to growth that occurs when cells attach to the substratum can be studied with small adhesive islands. Large numbers of these islands can be grouped together into arrays made up of various sizes, and the response of cells to incubation on these arrays allows the anchorage stimulus to be measured. Past work has shown that single isolated cells can be stimulated to proliferate under these circumstances quite as freely as they do in ordinary cultures, and that the maximum response is given by islands whose size is less than 5000 microns 2. This anchorage stimulus might be mediated by the cytoskeleton, which assembles rapidly around the points of attachment to the substratum. One possible approach to testing this hypothesis is to expose cells to islands of different shapes, and to search for common factors among the different arrangements of the cytoskeleton that these different islands cause. Circular islands induced a relatively disordered arrangement of actin fibres. The fibres were attached at one end to foci of vinculin, which sometimes became arranged in a ring around the margin of the island. Triangular islands showed a more orderly arrangement of actin, in three bands parallel to the sides. In this case, the vinculin accumulated at the apices. Long islands only 3 microns wide could also provide effective attachment for the cells. In this shape the actin accumulated in two bands 2 microns or more apart and up to 5 microns high, and the vinculin similarly collected in parallel interrupted bands along the margins of the island. The number of vinculin foci differed on these three different island shapes, and the total area of vinculin was more than three times greater on long islands than on circles or triangles of the same size. Despite these differences, all three different shapes of island were capable of inducing up to 100 microns 2 of vinculin foci in each cell. Round and triangular islands induced this maximum amount of vinculin when their size was 5000 microns 2. Linear islands induced the same amount when they were only 1000 microns 2. The effect of different shapes on total vinculin focal area was paralleled by their effects on growth. All three shapes could support a similar amount of proliferation. Round and triangular islands induced the maximum amount of proliferation when they were 5000 microns 2 in area, and linear islands when they were only 1000 microns 2.(ABSTRACT TRUNCATED AT 400 WORDS)
当细胞附着于基质时发生的生长刺激作用,可以通过小的黏附岛来进行研究。大量的这些岛可以组合在一起形成由各种大小组成的阵列,细胞在这些阵列上孵育的反应使得能够测量锚定刺激。过去的研究表明,在这种情况下,单个分离的细胞能够像在普通培养中一样自由地被刺激增殖,并且最大反应是由大小小于5000平方微米的岛产生的。这种锚定刺激可能是由细胞骨架介导的,细胞骨架会在附着于基质的点周围迅速组装。检验这一假设的一种可能方法是将细胞暴露于不同形状的岛,并在这些不同岛所导致的细胞骨架不同排列中寻找共同因素。圆形岛诱导肌动蛋白纤维形成相对无序的排列。这些纤维一端附着于纽蛋白焦点,纽蛋白焦点有时会围绕岛的边缘排列成环。三角形岛显示肌动蛋白有更有序的排列,呈三条与边平行的带。在这种情况下,纽蛋白聚集在顶点。仅3微米宽的长形岛也能为细胞提供有效的附着。在这种形状下,肌动蛋白聚集在两条相距2微米或更多、高达5微米的带中,纽蛋白同样沿着岛的边缘收集在平行的间断带中。这三种不同形状的岛上纽蛋白焦点的数量不同,长形岛上纽蛋白的总面积比相同大小的圆形或三角形岛大三倍多。尽管存在这些差异,但所有三种不同形状的岛都能够在每个细胞中诱导出多达100平方微米的纽蛋白焦点。当圆形和三角形岛的大小为5000平方微米时,诱导出这种最大量的纽蛋白。当线性岛只有1000平方微米时,诱导出相同的量。不同形状对纽蛋白总焦点面积的影响与其对生长的影响相似。所有三种形状都能支持相似数量的增殖。当圆形和三角形岛的面积为5000平方微米时,诱导出最大量的增殖,而线性岛在只有1000平方微米时诱导出最大量的增殖。(摘要截选至400字)