Hyperpolarization of the Plasma Membrane Potential Provokes Reorganization of the Actin Cytoskeleton and Increases the Stability of Adherens Junctions in Bovine Corneal Endothelial Cells in Culture
Hyperpolarization of the Plasma Membrane Potential Provokes Reorganization of the Actin Cytoskeleton and Increases the Stability of Adherens Junctions in Bovine Corneal Endothelial Cells in Culture
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DOI：
10.1002/cm.20416
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发表时间：
2009-12-01
影响因子：
--
通讯作者：
Chifflet, Silvia
中科院分区：
文献类型：
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作者：
Nin, Veronica;Hernandez, Julio A.;Chifflet, Silvia
在以前的工作中，我们表明，质膜电位（PMP）的去极化决定了文化中的不同上皮细胞的细胞骨架的重组，主要包括对细胞中心的外周肌动蛋白的重新分配，最终引起细胞间的破坏。鉴于这一证据，我们在这项研究中探讨了膜电位超极化对细胞骨架组织和粘附连接（AJ）的形态和培养的融合牛角膜内皮细胞的稳定性的可能影响。为此，超极化是通过用不可扩散的阳离子取代细胞外钠或通过将缬氨霉素掺入对照溶液来实现的。通过荧光显微镜图像的定量分析来评估细胞周围的肌动蛋白致密性。AJ的稳定性受到钙剥夺或温度降低的挑战。我们的研究结果表明，质膜超极化引起的压缩AJ-associated肌动蛋白丝对质膜和增加的稳定性AJs。我们还观察到，超极化程序确定在整个牛角膜内皮细胞的肌动蛋白细胞骨架类似的修改。连同我们以前的工作，这项研究的结果有助于的想法，在PMP的非兴奋性细胞的修改参与细胞的适应性反应，涉及重组的细胞骨架成分。细胞动力。Cytoskeleton 66：1087-1099，2009. (C)2009威利-利斯公司
In previous works we showed that the depolarization of the plasma membrane potential (PMP) determines a reorganization of the cytoskeleton of diverse epithelia in culture, consisting mainly of a reallocation of peripheral actin toward the cell center, ultimately provoking intercellular disruption. In view of this evidence, we explored in this study the possible effects of membrane potential hyperpolarization on the cytoskeletal organization and adherens junction (AJ) morphology and the stability of confluent bovine corneal endothelial cells in culture. For this purpose, hyperpolarization was achieved by substitution of extracellular sodium by nondiffusible cations or via the incorporation of valinomycin to the control solution. Actin compactness at the cell periphery was assessed by quantitative analysis of fluorescence microscopy images. The stability of the AJ was challenged by calcium deprivation or temperature decrease. Our results showed that plasma membrane hyperpolarization provokes a compaction of AJ-associated actin filaments toward the plasma membrane and an increase in the stability of the AJs. We also observed that the hyperpolarizing procedures determined similar modifications in the actin cytoskeleton of endothelial cells in whole bovine corneas. Together with our previous work, the results of this study contribute to the idea that modifications in the PMP of nonexcitable cells participate in cellular adaptive responses involving reorganization of cytoskeletal components. Cell Motil. Cytoskeleton 66: 1087-1099, 2009. (C) 2009 Wiley-Liss, Inc.