Simson R, Wallraff E, Faix J, Niewöhner J, Gerisch G, Sackmann E
Physics Department, Technische Universität München, Garching, Germany.
Biophys J. 1998 Jan;74(1):514-22. doi: 10.1016/S0006-3495(98)77808-7.
We have employed an interferometric technique for the local measurement of bending modulus, membrane tension, and adhesion energy of motile cells adhering to a substrate. Wild-type and mutant cells of Dictyostelium discoideum were incubated in a flow chamber. The flow-induced deformation of a cell near its adhesion area was determined by quantitative reflection interference contrast microscopy (RICM) and analyzed in terms of the elastic boundary conditions: equilibrium of tensions and bending moments at the contact line. This technique was employed to quantify changes caused by the lack of talin, a protein that couples the actin network to the plasma membrane, or by the lack of cortexillin I or II, two isoforms of the actin-bundling protein cortexillin. Cells lacking either cortexillin I or II exhibited reduced bending moduli of 95 and 160 k(B)T, respectively, as compared to 390 k(B)T, obtained for wild-type cells. No significant difference was found for the adhesion energies of wild-type and cortexillin mutant cells. In cells lacking talin, not only a strongly reduced bending modulus of 70 k(B)T, but also a low adhesion energy one-fourth of that in wild-type cells was measured.
我们采用了一种干涉测量技术来局部测量黏附在基质上的运动细胞的弯曲模量、膜张力和黏附能。将盘基网柄菌的野生型和突变型细胞置于流动腔中培养。通过定量反射干涉对比显微镜(RICM)确定细胞在其黏附区域附近因流动引起的变形,并根据弹性边界条件进行分析:接触线上的张力和弯矩平衡。该技术用于量化因缺乏将肌动蛋白网络与质膜连接的蛋白质踝蛋白,或因缺乏肌动蛋白成束蛋白皮层肌动蛋白I或II的两种同工型而导致的变化。与野生型细胞的390 k(B)T相比,缺乏皮层肌动蛋白I或II的细胞的弯曲模量分别降低至95和160 k(B)T。野生型细胞和皮层肌动蛋白突变型细胞的黏附能没有显著差异。在缺乏踝蛋白的细胞中,不仅测量到弯曲模量大幅降低至70 k(B)T,而且黏附能仅为野生型细胞的四分之一。