A Study of Electroporation of Cardiac Myocytes by Simultaneous Recording of Membrane current and Cellular Fluorescence
同时记录膜电流和细胞荧光的心肌细胞电穿孔研究
基本信息
- 批准号:12670046
- 负责人:
- 金额:$ 2.18万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2000
- 资助国家:日本
- 起止时间:2000 至 2001
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Strong electrical pulses produce membrane rupture that is associated with multiple pores (electroporation). Ion permeation through these pores can influence various cellular activities. In cardiac myocytes irregular inward currents (I_M) elicited by hyperpolarization has been assumed to represent electroporation. We examined it by simultaneously recording membrane currents and fluorescence of ethidium bromide (EB) that is membrane impermeable, binds with DNA and increases cellular fluorescence. Whole cell patch clamp technique and fluorescence microscope equipped with CCD camera were applied to rabbit ventricular myocytes supervising by K^+ -free solution with 5μM glibenclamide and 10 μg/ml EB. 40 s-negative square pulses from holding potential of -20 mV to potentials between -80 and -180 mV increased I_M with increase in hyperpolarization mainly by its conductance increase and it was associated with cellular EB fluorescence increase. The conductance produced by -80 and -1 6 0 mV steps were, 9.0±1.8 (n=14) and 66.2±25.4 pS/pF (n=7), and in these cells EB fluorescence increase was 0.062±0.005 and 0.321±0.075 in relative unit in the nuclei and 0.042±0.004 and 0.069±0.005 in the cytoplasm. EB fluorescence increased approximately in parallel with time integral of I_M, but nuclear EB fluorescence could rise also after I_M. LPC (10μM) enhanced I_M conductance to 103.0±21.8 pS/pF (n=5) and nuclear EB fluorescence to 0.34±0.12 in 5 min at -80 mV. Thus, hyperpolarization transiently ruptures membrane to produce pores to allow ethidium^+ (and Ca^<2+>) permeation and even more extensive membrane poration is produced by LPC at normal resting potential in cardiac myocytes.
强电脉冲产生与多个孔相关的膜破裂(电穿孔)。通过这些孔的离子渗透可以影响各种细胞活性。在心肌细胞中,由超极化引起的不规则内向电流(I_M)被认为代表了电穿孔。我们通过同时记录膜电流和溴化乙锭(EB)的荧光来研究它,溴化乙锭是膜不可渗透的,与DNA结合并增加细胞荧光。应用全细胞膜片钳技术和配有CCD摄像头的荧光显微镜观察含5μM格列本脲和10 μg/ml EB的无钾液对兔心室肌细胞的监控。从-20 mV到-80 ~-180 mV的40 s负方波脉冲使I_M随超极化的增加而增加,主要是通过增加I_M的电导而增加,并与细胞EB荧光的增加有关。-80 mV和-160 mV阶跃产生的电导分别为9.0±1.8(n=14)和66.2±25.4 pS/pF(n=7),细胞核EB荧光相对单位增加分别为0.062±0.005和0.321±0.075,细胞质EB荧光相对单位增加分别为0.042±0.004和0.069±0.005。EB荧光强度随I_M时间积分的增加而增加,但I_M后细胞核EB荧光强度也随之增加。LPC(10μM)在-80 mV时,5 min内使IM电导增加到103.0± 21.8pS/pF(n=5),核EB荧光增加到0.34±0.12。因此,在心肌细胞中,超极化会短暂地使膜破裂,产生孔,使乙锭^+(和Ca^2+)渗透,而LPC在正常静息电位下会产生更广泛的膜穿孔。
项目成果
期刊论文数量(22)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
渡邉 マキノ, 大地 陸男: "血管内皮細胞におけるCl^-電流および細胞内Ca^<2+>濃度の浸透圧による修飾"脈管学. 40. 205-209 (2000)
Makino Watanabe、Rikuo Daichi:“渗透压对血管内皮细胞中 Cl^- 电流和细胞内 Ca^2+ 浓度的修改”Angiology 40. 205-209 (2000)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Gupte SA, Okada T, Tateyama M, Ochi R: "Activation of (TxA_<2>/PGH_<2>) receptors and protein kinase C contribute to coronary dysfuction in supcroxide treated rat hearts."J Mol Cell Cardiol. 32. 937-946 (2000)
Gupte SA、Okada T、Tateyama M、Ochi R:“(TxA_2/PGH_2) 受体和蛋白激酶 C 的激活导致超氧化物处理的大鼠心脏中的冠状动脉功能障碍。”J Mol Cell Cardiol。
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- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Sonoda S, Ochi R: "Independent modulation of L-type Ca2+ channel in guinea pig ventricular cells by nitrendipine and isoproterenol."Jpn Heart J. 42. 77-86 (2002)
Sonoda S、Ochi R:“尼群地平和异丙肾上腺素对豚鼠心室细胞中 L 型 Ca2 通道的独立调节。”Jpn Heart J. 42. 77-86 (2002)
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
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大地陸男: "心血管系における細胞膜傷害"脈管学. 41. 13-22 (2001)
Rikuo Daichi:“心血管系统中的细胞膜损伤”血管学。41. 13-22 (2001)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Song YM, Ochi R: "Voltage-dependent increase of nuclear ethidium fluorescence in rabbit ventricular myocytes."Biophys J. 82. 96a-97a (2002)
Song YM、Ochi R:“兔心室肌细胞中核乙锭荧光的电压依赖性增加。”Biophys J. 82. 96a-97a (2002)
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- 影响因子:0
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OCHI Rikuo其他文献
OCHI Rikuo的其他文献
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{{ truncateString('OCHI Rikuo', 18)}}的其他基金
Study on mechanism of modulation of cardiac L-type Ca^<2+> channel by phosphorylation and Ca^<2+>
磷酸化和Ca^<2>调节心脏L型Ca^<2>通道的机制研究
- 批准号:
07457013 - 财政年份:1995
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Electrophysiological study of existence and regulation of chloride current in endothelial cells
内皮细胞氯电流存在及其调控的电生理学研究
- 批准号:
04454132 - 财政年份:1992
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Analysis of the Gating Mechanism of Cardiac Ca Channel by Long Recorder
长记录仪分析心脏Ca通道门控机制
- 批准号:
02670041 - 财政年份:1990
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
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