Yang W P, Onuma E K, Hui S W
Exp Cell Res. 1984 Nov;155(1):92-104. doi: 10.1016/0014-4827(84)90770-5.
C3H/10T1/2 mouse embryo fibroblasts were stimulated by a steady electric field ranging up to 15 V/cm. The percentage of spindle-shaped cells increased with the field strength and duration of the stimulation. These cells oriented preferentially with their long axis perpendicular to the field direction. A small percentage of the cells were found to move slightly toward the cathode during the course of electric stimulation. Although no apparent field-induced redistribution of fluorescent-labelled concanavalin A (conA) receptor along the cell periphery was observed, the bright perinuclear area appeared preferentially on the anode side. Correlative fluorescence and scanning electron microscopy (SEM) revealed no difference in the density of conA-gold microsphere labels on either side of the cell. The density of intramembranous particles on the E-face of the plasma membrane was 54% higher on the anode side than on the cathode side of the cell. The microfilament bundles were observed to be disrupted after 30 min of 10 V/cm stimulation by rhodamine phalloidin labelling of F-actin. The cell sensitivity to electric field-induced reorientation and cell shape changes was reduced by pretreatment with conA, and to a lesser extent, with succinyl conA or wheat germ agglutinin (WGA). ConA pretreatment alone also reduced the prominence of microfilament bundles. However, post-field lectin binding to the cell has no effect on cell recovery. It is possible that the generally flat 10T1/2 cells retract and realign in order to minimize the disruption of their membrane potential. The conA binding-mediated receptor-cytoskeletal linkage temporarily immobilizes the cell and inhibits subsequent field-induced shape changes.
C3H/10T1/2小鼠胚胎成纤维细胞受到高达15 V/cm的稳定电场刺激。纺锤形细胞的百分比随场强和刺激持续时间的增加而升高。这些细胞优先使其长轴垂直于场方向排列。在电刺激过程中,发现一小部分细胞会略微向阴极移动。尽管未观察到荧光标记的伴刀豆球蛋白A(conA)受体沿细胞周边出现明显的场诱导再分布,但明亮的核周区域优先出现在阳极侧。相关荧光和扫描电子显微镜(SEM)显示,细胞两侧的conA-金微球标记密度没有差异。细胞膜E面的膜内颗粒密度在细胞阳极侧比阴极侧高54%。通过用罗丹明鬼笔环肽标记F-肌动蛋白,观察到在10 V/cm刺激30分钟后微丝束被破坏。用conA预处理可降低细胞对电场诱导的重新定向和细胞形状变化的敏感性,琥珀酰conA或麦胚凝集素(WGA)预处理的影响程度较小。单独用conA预处理也会降低微丝束的突出程度。然而,场后凝集素与细胞的结合对细胞恢复没有影响。一般扁平的10T1/2细胞可能会收缩并重新排列,以尽量减少其膜电位的破坏。conA结合介导的受体-细胞骨架连接会暂时固定细胞,并抑制随后场诱导的形状变化。