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Shear-induced Volume Decrease in MDCK Cells

机译：剪切诱导的MDCK细胞体积减少

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Using a microfluidic cell volume sensor we measured the change in the cell volume of Madin-Darby Canine Kidney (MDCK) cells induced by shear stress. An increase in shear stress from 0.2 to 2.0 dyn/cmsup2/sup resulted in a volume decrease to a steady state volume ～ 20 – 30 % smaller than the initial resting cell volume. Independent experiments based on fluorescence quenching confirmed the volume reduction. This shear-induced cell shrinkage was irreversible on the time scale of the experiment (～ 30 min). Treatment of 0.1 µM Hgsup2+/sup significantly inhibited the volume decrease, suggesting that the shear-induced cell shrinkage is associated with water efflux through aquaporins. The volume decrease cannot be inhibited by 75 mM TEA, 100 µM DIDS, or 100 µM Gdsup3+/sup suggesting that volume reduction is not directly mediated by Ksup+/sup and Clsup-/supchannels that typically function during regulatory volume decrease (RVD), nor is it through cationic stretch-activated ion channels (SACs). The process also appears to be Casup2+/sup independent because it was insensitive to intracellular Casup2+/sup level. Since cell volume is determined by the intracellular water content, we postulate that the shear induced reductions in cell volume may arise from increased intracellular hydrostatic pressure as the cell is deformed under flow, which promotes the efflux of water. The increase in internal pressure in a deformable object under the flow is supported by the finite element mechanical model.
机译：使用微流控细胞体积传感器，我们测量了剪切应力诱导的Madin-Darby犬肾（MDCK）细胞的细胞体积变化。剪切应力从0.2 dyn / cm 2 增加到稳态时，体积减小到比初始静息池小20〜30％。基于荧光猝灭的独立实验证实了体积的减少。这种剪切诱导的细胞收缩在实验时间范围内（〜30分钟）是不可逆的。 0.1 µM Hg 2 + 的处理显着抑制了体积的减少，这表明剪切诱导的细胞收缩与水通道蛋白的水外排有关。 75 mM TEA，100 µM DIDS或100 µM Gd 3 + 不能抑制体积减小，这表明体积减小不是直接由K + 和Cl -通道，也不通过阳离子拉伸激活离子通道（SAC）起作用。该过程似乎也与Ca 2 + 无关，因为它对细胞内Ca 2 + 的水平不敏感。由于细胞的体积是由细胞内的水含量决定的，因此我们推测剪切诱导的细胞体积的减少可能是由于细胞在流动状态下变形而引起的细胞内静水压力的增加，从而促进了水的外流。流动条件下可变形物体内部压力的增加由有限元力学模型支持。

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