Cellular and Molecular Biophysics Lab, Department of Physics, University of South Florida, 4202 E. Fowler Ave., Tampa, FL 33620, USA.
J Membr Biol. 2012 Feb;245(2):97-105. doi: 10.1007/s00232-012-9418-6. Epub 2012 Feb 23.
Our previous studies have shown that a specially designed, so-called synchronization modulation electric field can entrain active transporter Na/K pumps in the cell membrane. This approach was previously developed in a study of single cells using a voltage clamp to monitor the pump currents. We are now expanding our study from isolated single cells to aggregated cells in a 3-dimensional cell matrix, through the use of a tissue slice from the rat heart. The slice is about 150 μm in thickness, meaning the slices contain many cell layers, resulting in a simplified 3-dimensional system. A fluorescent probe was used to identify the membrane potential and the ionic concentration gradients across the cell membrane. In spite of intrinsic cell-to-cell interactions and the difficulty in stimulating cell aggregation in the tissue slice, the oscillating electric field increased the intracellular fluorescent intensity, indicating elevation of the cell ionic concentration and hyperpolarization of the cell membrane. Blockage of these changes by ouabain confirmed that the results are directly related to Na/K pumps. These results along with the backward modulation indicate that the synchronization modulation electric field can influence the Na/K pumps in tissue cells of a 3-dimensional matrix and therefore hyperpolarize the cell membrane.
我们之前的研究表明,一种特殊设计的所谓同步调制电场可以使细胞膜中的主动转运体钠钾泵同步运转。该方法最初是在一项使用电压钳监测泵电流的单细胞研究中开发的。现在,我们通过使用来自大鼠心脏的组织切片,将研究从单个分离细胞扩展到三维细胞基质中的聚集细胞。该切片厚度约为 150μm,这意味着切片包含许多细胞层,形成了简化的三维系统。荧光探针用于识别细胞膜电位和跨细胞膜的离子浓度梯度。尽管存在固有细胞间相互作用以及在组织切片中刺激细胞聚集的困难,振荡电场增加了细胞内荧光强度,表明细胞离子浓度升高和细胞膜超极化。哇巴因阻断这些变化证实了这些结果与钠钾泵直接相关。这些结果以及反向调制表明,同步调制电场可以影响三维基质中组织细胞的钠钾泵,从而使细胞膜超极化。