Yvon Anne-Marie C, Walker Jonathan W, Danowski Barbara, Fagerstrom Carey, Khodjakov Alexey, Wadsworth Patricia
Department of Biology and Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, MA 01002, USA.
Mol Biol Cell. 2002 Jun;13(6):1871-80. doi: 10.1091/mbc.01-11-0539.
The reorientation of the microtubule organizing center during cell migration into a wound in the monolayer was directly observed in living wound-edge cells expressing gamma-tubulin tagged with green fluorescent protein. Our results demonstrate that in CHO cells, the centrosome reorients to a position in front of the nucleus, toward the wound edge, whereas in PtK cells, the centrosome lags behind the nucleus during migration into the wound. In CHO cells, the average rate of centrosome motion was faster than that of the nucleus; the converse was true in PtK cells. In both cell lines, centrosome motion was stochastic, with periods of rapid motion interspersed with periods of slower motion. Centrosome reorientation in CHO cells required dynamic microtubules and cytoplasmic dynein/dynactin activity and could be prevented by altering cell-to-cell or cell-to-substrate adhesion. Microtubule marking experiments using photoactivation of caged tubulin demonstrate that microtubules are transported in the direction of cell motility in both cell lines but that in PtK cells, microtubules move individually, whereas their movement is more coherent in CHO cells. Our data demonstrate that centrosome reorientation is not required for directed migration and that diverse cells use distinct mechanisms for remodeling the microtubule array during directed migration.
在表达绿色荧光蛋白标记的γ-微管蛋白的活伤口边缘细胞中,直接观察到细胞迁移到单层伤口时微管组织中心的重新定向。我们的结果表明,在CHO细胞中,中心体重新定向到细胞核前方、朝向伤口边缘的位置,而在PtK细胞中,在迁移到伤口的过程中,中心体落后于细胞核。在CHO细胞中,中心体运动的平均速率比细胞核快;在PtK细胞中则相反。在这两种细胞系中,中心体运动都是随机的,快速运动期与慢速运动期交替出现。CHO细胞中的中心体重新定向需要动态微管和细胞质动力蛋白/动力蛋白激活蛋白的活性,并且可以通过改变细胞间或细胞与底物的黏附来阻止。使用笼化微管蛋白光激活的微管标记实验表明,在这两种细胞系中微管都沿细胞运动方向运输,但在PtK细胞中,微管单独移动,而在CHO细胞中它们的移动更连贯。我们的数据表明,定向迁移不需要中心体重新定向,并且不同的细胞在定向迁移过程中使用不同的机制来重塑微管阵列。