Plasma membrane voltage changes during nanosecond pulsed electric field exposure
Plasma membrane voltage changes during nanosecond pulsed electric field exposure
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DOI：
10.1529/biophysj.105.072777
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发表时间：
2006-05-01
影响因子：
3.4
通讯作者：
Kolb, JF
中科院分区：
文献类型：
--
作者：
Frey, W;White, JA;Kolb, JF
研究了持续时间为60 ns、最大电场强度接近100 kV/cm （100 V/细胞直径）的纳秒脉冲电场对Jurkat细胞膜电位的响应。用快速电压敏感染料ANNINE-6对Jurkat细胞膜进行染色，该染料具有亚纳秒电压响应时间。用可调谐激光脉冲激发该染料，获得了5ns的时间分辨率。激光脉冲与外加电场同步，在曝光前、期间和之后记录图像。当Jurkat电池暴露在脉冲下时，电池阳极极膜上的电压在15 ns后达到1.6 V的值，几乎是电穿孔通常所需电压水平的两倍。在同一时间段内，面向阴极一侧膜上的电压仅达到0.6 V，这表明两个相对细胞半球膜上的传导机制存在强烈的不对称性。在质膜上的0.6 - 1.6 V的小电压降表明，几乎整个施加的10 V/ μ m的电场都渗透到细胞内部和每个细胞器中。
The change in the membrane potential of Jurkat cells in response to nanosecond pulsed electric fields was studied for pulses with a duration of 60 ns and maximum field strengths of similar to 100 kV/cm (100 V/cell diameter). Membranes of Jurkat cells were stained with a fast voltage-sensitive dye, ANNINE-6, which has a subnanosecond voltage response time. A temporal resolution of 5 ns was achieved by the excitation of this dye with a tunable laser pulse. The laser pulse was synchronized with the applied electric field to record images at times before, during, and after exposure. When exposing the Jurkat cells to a pulse, the voltage across the membrane at the anodic pole of the cell reached values of 1.6 V after 15 ns, almost twice the voltage level generally required for electroporation. Voltages across the membrane on the side facing the cathode reached values of only 0.6 V in the same time period, indicating a strong asymmetry in conduction mechanisms in the membranes of the two opposite cell hemispheres. This small voltage drop of 0.6 - 1.6 V across the plasma membrane demonstrates that nearly the entire imposed electric field of 10 V/mu m penetrates into the interior of the cell and every organelle.