Following a central transcorneal circular freeze injury, organ-cultured rat corneal endothelial cells surrounding the wound reorganize peripheral actin bands into stress fibers and migrate individually into the wound. To ascertain the significance of this rearrangement relative to morphological changes accompanying migration and wound repair, some tissues were incubated overnight in 4 μM TRITC-conjugated phalloidin to stabilize actin and prevent its reorganization. After a freeze injury to the endothelium tissues were histologically observed at 24 h post-wounding and demonstrated that despite a lack of actin organization, cells responding to the injury appeared morphologically similar to their control counterparts. Tissues cultivated in the presence of either cytochalasin B (CB), soybean agglutinin (SBA), or fluorouracil (FU) and also exhibited actin cytoskeletal disruption. Under these conditions, migration continued despite the absence of detectable stress fibers. For SBA-, CB-, and FU-treated tissues, wound repair did not significantly differ from control preparations although FU-treated tissues showed a slower repair. Electron micrographs confirmed an absence of stress fibers in migrating cells treated with any of these agents. Tissues were also treated with ML 141 and EY294002 to inhibit the cdc-42 and PI-3K pathways, respectively. While cell movement still occurred in the presence of ML 141, migration into the wound was greatly restricted in the presence of EY294002. These results indicate that rat corneal endothelial cell movement in situ does not require actin reorganization into stress fibers, but the functioning of the PI-3K pathway is indispensable for their migration along the basement membrane during wound repair.

中文翻译：

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基底膜伤口修复过程中大鼠角膜内皮细胞迁移更多地依赖于 PI-3K 通路而不是 cdc-42 通路或肌动蛋白应力纤维

在中央经角膜圆形冷冻损伤后，伤口周围器官培养的大鼠角膜内皮细胞将外周肌动蛋白带重组为应力纤维并单独迁移到伤口中。为了确定这种重排相对于伴随迁移和伤口修复的形态变化的重要性，将一些组织在 4 μM TRITC 偶联的鬼笔环肽中孵育过夜，以稳定肌动蛋白并防止其重组。在受伤后 24 小时组织学观察内皮组织冷冻损伤后，证明尽管缺乏肌动蛋白组织，但对损伤有反应的细胞在形态上与对照细胞相似。在细胞松弛素 B (CB)、大豆凝集素 (SBA)、或氟尿嘧啶 (FU) 并且还表现出肌动蛋白细胞骨架破坏。在这些条件下，尽管没有可检测到的应力纤维，迁移仍在继续。对于 SBA、CB 和 FU 处理的组织，伤口修复与对照制剂没有显着差异，尽管 FU 处理的组织显示出较慢的修复。电子显微照片证实在用任何这些试剂处理的迁移细胞中不存在应力纤维。组织也用 ML 141 和 EY294002 处理以分别抑制 cdc-42 和 PI-3K 通路。虽然在 ML 141 存在下仍会发生细胞运动，但在 EY294002 存在下，向伤口的迁移受到极大限制。这些结果表明，大鼠角膜内皮细胞原位运动不需要肌动蛋白重组为应力纤维，