Department of Physiology and Biophysics, Dalhousie University, Halifax, NS, B3H 1X5, Canada.
J Membr Biol. 2010 Dec;238(1-3):69-80. doi: 10.1007/s00232-010-9320-z. Epub 2010 Nov 21.
Electroporation induced by high-strength electrical fields has long been used to investigate membrane properties and facilitate transmembrane delivery of molecules and genes for research and clinical purposes. In the heart, electric field-induced passage of ions through electropores is a factor in defibrillation and postshock dysfunction. Voltage-clamp pulses can also induce electroporation, as exemplified by findings in earlier studies on rabbit ventricular myocytes: Long hyperpolarizations to ≤-110 mV induced influx of marker ethidium and irregular inward currents that were as large with external NMDG(+) as Na(+). In the present study, guinea pig ventricular myocytes were bathed with NMDG(+), Na(+) or NMDG(+) + La(3+) solution (36°C) and treated with five channel blockers. Hyperpolarization of myocytes in NMDG(+) solution elicited an irregular inward current (I (ep)) that reversed at -21.5 ± 1.5 mV. In myocytes hyperpolarized with 200-ms steps every 30 s, I (ep) occurred in "episodes" that lasted for one to four steps. Boltzmann fits to data on the incidence of I (ep) per experiment indicate 50% incidence at -129.7 ± 1.4 mV (Na(+)) and -146.3 ± 1.6 mV (NMDG(+)) (slopes ≈-7.5 mV). I (ep) amplitude increased with negative voltage and was larger with Na(+) than NMDG(+) (e.g., -2.83 ± 0.34 vs. -1.40 ± 0.22 nA at -190 mV). La(3+) (0.2 mM) shortened episodes, shifted 50% incidence by -35 mV and decreased amplitude, suggesting that it inhibits opening/promotes closing of electropores. We compare our findings with earlier ones, especially in regard to electropore selectivity. In the Appendix, relative permeabilities and modified excluded-area theory are used to derive estimates of electropore diameters consistent with reversal potential -21.5 mV.
电场诱导的电穿孔长期以来一直被用于研究细胞膜特性,并促进分子和基因的跨膜传递,用于研究和临床目的。在心脏中,电场诱导的离子通过电孔的传递是除颤和电击后功能障碍的一个因素。电压钳脉冲也可以诱导电穿孔,这在前些年对兔心室肌细胞的研究中得到了例证:长时程超极化至≤-110 mV 会引起指示剂 ethidium 的内流和不规则内向电流,而用外源性 NMDG(+) 取代 Na(+) 时,这种电流会变得更大。在本研究中,豚鼠心室肌细胞用 NMDG(+)、Na(+)或 NMDG(+) + La(3+)溶液(36°C)孵育,并使用 5 种通道阻断剂处理。在 NMDG(+)溶液中,心肌细胞的超极化会引起不规则内向电流(I (ep)),该电流在-21.5 ± 1.5 mV 处反转。在每 30 秒用 200-ms 步长超极化的心肌细胞中,I (ep)会以持续 1 到 4 个步长的“发作”形式发生。Boltzmann 拟合实验中 I (ep)发生率的数据表明,在 50%的实验中,I (ep)的发生率在-129.7 ± 1.4 mV(Na(+))和-146.3 ± 1.6 mV(NMDG(+))(斜率≈-7.5 mV)。I (ep)的幅度随负电压的增加而增加,且在 Na(+)中的幅度大于 NMDG(+)(例如,在-190 mV 时,-2.83 ± 0.34 nA 比-1.40 ± 0.22 nA)。La(3+)(0.2 mM)缩短了发作时间,将 50%的发生率移向-35 mV,并降低了幅度,表明它抑制了电孔的开放/促进了电孔的关闭。我们将我们的发现与早期的发现进行了比较,特别是在电孔选择性方面。附录中,相对渗透率和修正的排除面积理论用于推导与反转电位-21.5 mV 一致的电孔直径估计值。