Cell-substrate impedance analysis of epithelial cell shape and micromotion upon challenge with bacterial proteins that perturb extracellular matrix and cytoskeleton
Cell-substrate impedance analysis of epithelial cell shape and micromotion upon challenge with bacterial proteins that perturb extracellular matrix and cytoskeleton
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DOI：
10.1016/s0167-7012(98)00083-9
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发表时间：
1998-11-01
影响因子：
2.2
通讯作者：
Ellen, RP
中科院分区：
文献类型：
--
作者：
Ko, KSC;Lo, CM;Ellen, RP
研究宿主细胞-微生物串扰的本质需要更灵敏的方法来分析细胞对感染或感染因子的毒性因子的实时反应。在这里，我们介绍了新的细胞-基质阻抗传感（ECIS）的分析应用，以量化细胞形态和微运动在响应细菌挑战时的变化。用密螺旋体（Treponema denticola）的外膜（OM）提取物、纯化的95 kDa凝乳胰蛋白酶和主要表面蛋白（Msp）攻击上皮单层。通过模型计算分析和区分阻抗变化为结电阻（R-b）或平均细胞-衬底分离(h)。用阻抗波动和归一化方差比来量化微动。OM和95 kDa蛋白酶使阻抗迅速下降。Msp对R-b和h没有影响，但有助于减少微动。这些新的ECIS分析应用可用于研究任何影响细胞连接、细胞骨架完整性、细胞形状或与细胞外基质接触的病原体的产毒活性。(C) 1998 Elsevier Science BN。版权所有。
Research into the nature of host cell-microbial cross-talk needs more sensitive methods to analyze real time cellular reactions to infection or to toxic factors from infecting agents. Here, we introduce novel analytical applications of electrical cell-substrate impedance sensing (ECIS) to quantify changes in cell morphology and micromotion in response to bacterial challenge. Epithelial monolayers were challenged with an outer membrane (OM) extract of Treponema denticola, its purified 95 kDa chymotrypsin, and major surface protein (Msp). Impedance changes were analyzed and discriminated through model calculation as either junctional resistance (R-b) or average cell-substrate separation (h). Fluctuations in impedance and the normalized variance ratio were used to quantify micromotion. The OM and the 95 kDa protease caused a rapid drop in impedance. Msp had no effect on R-b and h, but contributed to the reduction in micromotion. These new analytical applications of ECIS can be used to study toxigenic activities of any pathogen that affects cell junctions, cytoskeletal integrity, cell shape, or contact with extracellular matrix. (C) 1998 Elsevier Science BN. All rights reserved.