ELECTRIC-FIELD DIRECTED CELL-SHAPE CHANGES, DISPLACEMENT, AND CYTOSKELETAL REORGANIZATION ARE CALCIUM DEPENDENT
ELECTRIC-FIELD DIRECTED CELL-SHAPE CHANGES, DISPLACEMENT, AND CYTOSKELETAL REORGANIZATION ARE CALCIUM DEPENDENT
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DOI：
10.1083/jcb.106.6.2067
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发表时间：
1988-06-01
影响因子：
7.8
通讯作者：
HUI, SW
中科院分区：
文献类型：
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作者：
ONUMA, EK;HUI, SW
C3H/10T1/2小鼠胚胎成纤维细胞受到高达10 V/cm的稳定电场刺激。一些细胞伸长并垂直于磁场方向排列。钙通道阻滞剂D-600和钙调素拮抗剂三氟拉嗪抑制了向阴极的优先位置移动。罗丹明-phalloidin标记的肌动蛋白丝显示了电场诱导的应力纤维模式的紊乱，当在缺钙缓冲液或含有钙拮抗剂CoCl2、钙通道阻滞剂D-600或钙调素拮抗剂三氟拉嗪的缓冲液中进行刺激时，这种紊乱会减少。钙离子载体A23187处理也有类似的效果，但D-600的存在并没有减少应力纤维的破坏。钙敏感光蛋白aequorin用于监测细胞内游离钙的变化。电刺激使钙增加到微摩尔范围。这种增加被缺钙缓冲液或CoCl2抑制，并被D-600降低。一个钙依赖的机制被提出来解释观察到的场定向细胞形状变化，优先取向和位移。
C3H/10T1/2 mouse embryo fibroblasts were stimulated by a steady electric field ranging up to 10 V/cm. Some cells elongated and aligned perpendicular to the field direction. A preferential positional shift toward the cathode was observed which was inhibited by the calcium channel blocker D-600 and the calmodulin antagonist trifluoperazine. Rhodamine-phalloidin labeling of actin filaments revealed a field-induced disorganization of the stress fiber pattern, which was reduced when stimulation was conducted in calcium-depleted buffer or in buffer containing calcium antagonist CoCl2, calcium channel blocker D-600, or calmodulin antagonist trifluoperazine. Treatment with calcium ionophore A23187 had similar effects, except that the presence of D-600 did not reduce the stress fiber disruption. The calcium-sensitive photoprotein aequorin was used to monitor changes in intracellular-free calcium. Electric stimulation caused an increased of calcium to the micromolar range. This increase was inhibited by calcium-depleted buffer or by CoCl2, and was reduced by D-600. A calcium-dependent mechanism is proposed to explain the observed field-directed cell shape changes, preferential orientation, and displacement.