Cell stimulation and calcium mobilization by picosecond electric pulses.

Cell stimulation and calcium mobilization by picosecond electric pulses.
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DOI:
10.1016/j.bioelechem.2015.05.013
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发表时间:
2015-10
影响因子:
5
通讯作者:
Pakhomov, Andrei G.
Pakhomov, Andrei G.
中科院分区:
化学2区
文献类型:
--
作者:
Semenov, Iurii;Xiao, Shu;Kang, Dongkoo;Schoenbach, Karl H.;Pakhomov, Andrei G.

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我们测试了比通道激活时间短得多的皮秒电脉冲(psEP;190 kV/cm,500 ps,50% 高度)是否可以激活电压门控(VG)通道。通过 Fura-2 比率成像技术监测 GH3 和 NG108 细胞(表达多种类型的 VG 钙通道 VGCC)和 CHO 细胞(不表达 VGCC)中的胞质 Ca2+。 1 kHz 下高达 100 psEP 的训练在 CHO 细胞中没有引起任何反应。然而,即使是单个 psEP 也会显着增加 GH3(增加 114+/-48 nM)和 NG108 细胞(增加 6 +/-1.1 nM)中的 Ca2+。 100 psEP 的序列将响应分别放大至 379+/-33 nM 和 719+/-315 nM。 Ca2+ 反应在 2-15 秒内达到峰值,并在 100 秒以上恢复;它们被维拉帕米和 ω-芋螺毒素抑制 80-100%,但用 N-甲基-D-葡萄糖胺取代 Na+ 则不能抑制它们。在不含 Ca2+ 的培养基中对 psEP 没有反应,但添加外部 Ca2+ 即使 10 秒后也能引起 Ca2+ 反应。我们得出的结论是,短至 500 ps 的电刺激可以通过不涉及传统电穿孔、加热(每 psEP 低于 0.06 °K)或通过打开 VG Na+ 通道进行膜去极化的机制来导致 VGCC 持久打开。
We tested if picosecond electric pulses (psEP; 190 kV/cm, 500 ps at 50% height), which are much shorter than channel activation time, can activate voltage-gated (VG) channels. Cytosolic Ca2+ was monitored by Fura-2 ratiometric imaging in GH3 and NG108 cells (which express multiple types of VG calcium channels, VGCC), and in CHO cells (which express no VGCC). Trains of up to 100 psEP at 1 kHz elicited no response in CHO cells. However, even a single psEP significantly increased Ca2+ in both GH3 (by 114+/−48 nM) and NG108 cells (by 6 +/−1.1 nM). Trains of 100 psEP amplified the response to 379+/−33 nM and 719+/−315 nM, respectively. Ca2+ responses peaked within 2–15 s and recovered for over 100 s; they were 80–100% inhibited by verapamil and ω-conotoxin, but not by the substitution of Na+ with N-methyl-D-glucamine. There was no response to psEP in Ca2+-free medium, but adding external Ca2+ even 10 s later evoked Ca2+ response. We conclude that electrical stimuli as short as 500 ps can cause long-lasting opening of VGCC by a mechanism which does not involve conventional electroporation, heating (which was under 0.06 °K per psEP), or membrane depolarization by opening of VG Na+ channels.
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