Electroporation by subnanosecond pulses.
Electroporation by subnanosecond pulses.
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DOI：
10.1016/j.bbrep.2016.05.002
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发表时间：
2016-07
影响因子：
2.7
通讯作者：
Pakhomov AG
中科院分区：
文献类型：
--
作者：
Semenov I;Xiao S;Pakhomov AG
微秒和纳秒刺激对细胞膜的电穿透作用已经被广泛研究，而皮秒电脉冲(PsEP)的作用基本上还没有被探索。我们利用全细胞膜片钳和Di-8-ANEPPS电压敏感染料测量方法，研究了500npS刺激对大鼠海马神经元(RHN)、NG108和CHO细胞质膜的影响。即使是190千伏/厘米的500ps脉冲也会增加膜电导并使细胞去极化。这些效应通过施加短暂的psEP脉冲(5-125个脉冲)而被放大，而脉冲传递速率(8 GHz-1 GHz)起到的作用很小。PSEP处理的细胞在负膜电位下表现出较大的内向电流，而在正电位下表现出较小的电导变化或没有变化。25个脉冲的1 kHz脉冲使RHN和−108细胞的全细胞电导在(−100)-(NG60)mV范围内增加到22-26nS(分别从3和0.7nS)，但在CHO细胞中仅增加到5nS(从0.3nS)。电导的增加在约2分钟内可逆。这种细胞通透性模式具有向内整流和缓慢恢复的特点，与早先报道的60 ns和600 ns脉冲的效应相似，表明尽管膜充电机制不同，但结构膜重排相似。在190千伏/厘米的500-ps脉冲电穿孔哺乳动物细胞。在负电位时，细胞失去膜电位，电导增加。通过增加脉冲次数而不是通过增加速率来增强透过性。膜的重新密封需要几分钟。
Electropermeabilization of cell membranes by micro- and nanosecond-duration stimuli has been studied extensively, whereas effects of picosecond electric pulses (psEP) remain essentially unexplored. We utilized whole-cell patch clamp and Di-8-ANEPPS voltage-sensitive dye measurements to characterize plasma membrane effects of 500 ps stimuli in rat hippocampal neurons (RHN), NG108, and CHO cells. Even a single 500-ps pulse at 190 kV/cm increased membrane conductance and depolarized cells. These effects were augmented by applying brief psEP bursts (5–125 pulses), whereas the rate of pulse delivery (8 Hz–1 kHz) played little role. psEP-treated cells displayed large inward current at negative membrane potentials but modest or no conductance changes at positive potentials. A 1-kHz burst of 25 pulses increased the whole-cell conductance in the range (−100)–(−60) mV to 22–26 nS in RHN and NG108 cells (from 3 and 0.7 nS, respectively), but only to 5 nS in CHO (from 0.3 nS). The conductance increase was reversible within about 2 min. Such pattern of cell permeabilization, with characteristic inward rectification and slow recovery, was similar to earlier reported effects of 60- and 600-ns pulses, pointing to the similarity of structural membrane rearrangements in spite of a different membrane charging mechanism. 500-ps pulses at 190 kV/cm electroporate mammalian cells. Cells lose membrane potential and increase conductance at negative potentials. Permeabilization is enhanced by increasing pulse number but not the rate. Membrane resealing takes minutes.