Role of Rho GTPase in astrocyte morphology and migratory response during in vitro wound healing
Role of Rho GTPase in astrocyte morphology and migratory response during in vitro wound healing
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DOI：
10.1111/j.1471-4159.2005.03443.x
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发表时间：
2005-12-01
影响因子：
4.7
通讯作者：
Ahnert-Hilger, G
中科院分区：
文献类型：
--
作者：
Höltje, M;Hoffmann, A;Ahnert-Hilger, G
小 Rho GTP 酶是多种细胞中细胞骨架的关键调节因子。 Rho 功能介导形态变化以及运动活动。利用新生小鼠建立的星形胶质细胞培养物，我们研究了 Rho 在星形胶质细胞星状形成过程形成中的作用。使用划伤伤口模型，我们研究了 Rho 对伤口愈合过程中各种形态和功能变量（例如星状和迁移活动）的影响。 C3 蛋白广泛用于研究细胞 Rho 功能。此外，源自肉毒杆菌(C3(bot))的C3被认为选择性地促进神经元再生。由于后者需要神经元和神经胶质细胞的平衡活性，因此必须仔细考虑C3蛋白对星形胶质细胞等神经胶质细胞的影响。低纳摩尔浓度的 C3 蛋白显着促进过程生长并增加过程分支。除了 ADP-核糖基化导致 Rho 酶失活之外，各种 Rho GTP 酶的蛋白质水平的变化也可能有助于观察到的效果。此外，用 C3(bot) 孵育划伤的星形胶质细胞培养物可加速伤口愈合。通过用选择性抑制剂 Y27632 抑制 Rho 下游效应器 ROCK，我们能够证明加速伤口闭合是由于极化过程形成增强和星形胶质细胞向病变部位迁移活动增加所致。这些结果表明，Rho 负向调节损伤后星形胶质细胞的生长和迁移反应，并且纳摩尔浓度的 C3(bot) 对其失活可促进星形胶质细胞迁移。
Small Rho GTPases are key regulators of the cytoskeleton in a great variety of cells. Rho function mediates morphological changes as well as locomotor activity. Using astrocyte cultures established from neonatal mice we investigated the role of Rho in process formation during astrocyte stellation. Using a scratch-wound model, we examined the impact of Rho on a variety of morphological and functional variables such as stellation and migratory activity during wound healing. C3 proteins are widely used to study cellular Rho functions. In addition, C3 derived from Clostridium botulinum (C3(bot)) is considered selectively to promote neuronal regeneration. Because the latter requires a balanced activity of neurones and glial cells, the effects of C3 protein on glial cells such as astrocytes have to be considered carefully. Low nanomolar concentrations of C3 proteins significantly promoted process outgrowth and increased process branching. Besides enzymatic inactivation of Rho by ADP-ribosylation, changes in protein levels of the various Rho GTPases may also contribute to the observed effects. Furthermore, incubation of scratch-wounded astrocyte cultures with C3(bot) accelerated wound healing. By inhibiting the Rho downstream effector ROCK with the selective inhibitor Y27632 we were able to demonstrate that the accelerated wound closure resulted from both enhanced polarized process formation and increased migratory activity of astrocytes into the lesion site. These results suggest that Rho negatively regulates astrocytic process growth and migratory responses after injury and that its inactivation by C3(bot) in nanomolar concentrations promotes astrocyte migration.