Surface Plasmon Resonance Monitoring of Cell Monolayer Integrity: Implication of Signaling Pathways Involved in Actin-Driven Morphological Remodeling.
Surface Plasmon Resonance Monitoring of Cell Monolayer Integrity: Implication of Signaling Pathways Involved in Actin-Driven Morphological Remodeling.
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DOI：
10.1007/s12195-008-0028-4
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发表时间：
2008-12-01
影响因子：
2.8
通讯作者：
Grandbois, Michel
中科院分区：
文献类型：
--
作者：
Cuerrier, Charles M.;Chabot, Vincent;Vigneux, Sylvain;Aimez, Vincent;Escher, Emanuel;Gobeil, Fernand, Jr.;Charette, Paul G.;Grandbois, Michel
单个细胞中发生的形态变化在很大程度上影响了各种细胞层的生理功能。上皮和内皮的屏障功能的控制是高度取决于细胞形态和细胞层完整性的过程的一个主要例子。在这里，我们将表面等离子体共振（SPR）技术应用于由血管紧张素II刺激的上皮细胞单层的细胞活性的定量。 SPR信号的分析显示了用血管紧张素II激活细胞激活后可再现浓度依赖性双相反应。进行相位对比和共聚焦显微镜成像，以将SPR信号与分子和全局形态重塑联系起来。观察到SPR信号与快速细胞收缩和随后通过相比显微镜观察到的细胞扩散有关。此外，血管紧张素II刺激后通过共聚焦显微镜观察到肌动蛋白的时间再分布，也与SPR信号变化一致。肌动蛋白 - 肌球蛋白驱动的细胞收缩中涉及的信号通路的调节证实了肌动蛋白结构在SPR反应中的直接含义。此外，我们表明与血管紧张素II刺激相关的细胞内钙动员不会产生任何显着的SPR信号变化。总而言之，我们的结果表明，SPR是一种研究细胞活性和分子机制的快速无标记方法，该方法与细胞单层的完整性有关，与相关的细胞形态变化有关。
Morphological changes occurring in individual cells largely influence the physiological functions of various cell layers. The control of barrier function of epithelia and endothelia is a prime example of processes highly dependent on cellular morphology and cell layer integrity. Here, we applied the surface plasmon resonance (SPR) technique to the quantification of cellular activity of an epithelial cell monolayer stimulated by angiotensin II. The analysis of the SPR signal shows reproducible concentration-dependent biphasic responses after cell activation with angiotensin II. Phase-contrast and confocal microscopy imaging was performed to link the SPR signal to molecular and global morphological remodeling. The SPR signal was observed to be in relation with the rapid cell contraction and the subsequent cell spreading observed by phase-contrast microscopy. Additionally, the temporal redistribution of actin, observed by confocal microscopy after angiotensin II stimulation, was also found to be consistent with the SPR signal variation. The modulation of signaling pathways involved in actin-myosin driven cell contraction confirms the direct implication of actin structures in the SPR response. Additionally, we show that the intracellular calcium mobilization associated with angiotensin II stimulation did not produce any significant SPR signal variation. Altogether, our results demonstrate that SPR is a rapid label-free method to study cellular activity and molecular mechanisms implicated in the modulation of the integrity of a cell monolayer in relation to cytoskeleton remodeling with associated cell morphological changes.
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影响因子：
3.7
作者：
Auger-Messier, M;Turgeon, ES;Guillemette, G
通讯作者：
Guillemette, G
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Siess, W
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