Delanoë-Ayari H, Iwaya S, Maeda Y T, Inose J, Rivière C, Sano M, Rieu J-P
Université de Lyon, F-6900, France, Université Lyon 1,CNRS UMR 5586, F-69622 Villeurbanne Cedex, France.
Cell Motil Cytoskeleton. 2008 Apr;65(4):314-31. doi: 10.1002/cm.20262.
The distribution of forces exerted by migrating Dictyostelium amebae at different developmental stages was measured using traction force microscopy. By using very soft polyacrylamide substrates with a high fluorescent bead density, we could measure stresses as small as 30 Pa. Remarkable differences exist both in term of the magnitude and distribution of forces in the course of development. In the vegetative state, cells present cyclic changes in term of speed and shape between an elongated form and a more rounded one. The forces are larger in this first state, especially when they are symmetrically distributed at the front and rear edge of the cell. Elongated vegetative cells can also present a front-rear asymmetric force distribution with the largest forces in the crescent-shaped rear of the cell (uropod). Pre-aggregating cells, once polarized, only present this last kind of asymmetric distribution with the largest forces in the uropod. Except for speed, no cycle is observed. Neither the force distribution of pre-aggregating cells nor their overall magnitude are modified during chemotaxis, the later being similar to the one of vegetative cells (F(0) approximately 6 nN). On the contrary, both the force distribution and overall magnitude is modified for the fast moving aggregating cells. In particular, these highly elongated cells exert lower forces (F(0) approximately 3 nN). The location of the largest forces in the various stages of the development is consistent with the myosin II localization described in the literature for Dictyostelium (Yumura et al.,1984. J Cell Biol 99:894-899) and is confirmed by preliminary experiments using a GFP-myosin Dictyostelium strain.
利用牵引力显微镜测量了不同发育阶段迁移的盘基网柄菌变形虫所施加力的分布情况。通过使用具有高荧光珠密度的非常柔软的聚丙烯酰胺底物,我们能够测量低至30帕斯卡的应力。在发育过程中,力的大小和分布方面均存在显著差异。在营养状态下,细胞在伸长形态和更圆润形态之间呈现速度和形状的周期性变化。在这种初始状态下力更大,尤其是当它们对称分布在细胞的前缘和后缘时。伸长的营养细胞也可以呈现前后不对称的力分布,在细胞新月形的后部(尾足)力最大。预聚集细胞一旦极化,仅呈现最后这种不对称分布,在尾足处力最大。除了速度外,未观察到周期性变化。在趋化作用期间,预聚集细胞的力分布及其总体大小均未改变,后者与营养细胞的相似(F(0)约为6纳牛顿)。相反,快速移动的聚集细胞的力分布和总体大小均发生了改变。特别是,这些高度伸长的细胞施加的力较低(F(0)约为3纳牛顿)。发育各阶段最大力的位置与文献中描述的盘基网柄菌肌球蛋白II的定位一致(Yumura等人,1984年。《细胞生物学杂志》99:894 - 899),并且通过使用绿色荧光蛋白 - 肌球蛋白盘基网柄菌菌株的初步实验得到了证实。