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原代培养大鼠主动脉平滑肌细胞中的低电压激活钙电流

Low-voltage-activated calcium current in rat aorta smooth muscle cells in primary culture.

作者信息

Akaike N, Kanaide H, Kuga T, Nakamura M, Sadoshima J, Tomoike H

机构信息

Research Institute of Angiocardiology, Kyushu University, Fukuoka, Japan.

出版信息

J Physiol. 1989 Sep;416:141-60. doi: 10.1113/jphysiol.1989.sp017754.

DOI:10.1113/jphysiol.1989.sp017754
PMID:2558173
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1189208/
Abstract
  1. Electrical and pharmacological properties of the low-voltage-activated Ca2+ current (ICa, LVA) in rat aorta smooth muscle cells (SMC) in primary culture were examined, particularly in comparison with the high-voltage-activated Ca2+ current (ICa, HVA). Both types of Ca2+ currents were recorded in external solution containing 20 mM-Ca2+, using the whole-cell voltage-clamp technique. 2. ICa, LVA was evoked by step depolarizations to potentials more positive than -60 mV from a holding potential of -100 mV, and reached a peak in the current-voltage (I-V) relationship around -30 mV. ICa, HVA was activated at -20 mV, and reached a peak at +20 mV. 3. The intracellular dialysis of 5 mM-F- irreversibly suppressed ICa, HVA, with time, while it has little effect on the ICa, LVA. The ICa, LVA could be separated from the ICa, HVA by either selecting the holding and test potential levels or by perfusing intracellularly with F-. 4. The ratio of peak amplitude of Ba2+, Sr2+ and Ca2+ currents in the respective I-V relationship was 1.6:1.2:1.0 for high-voltage-activated Ca2+ channels and was 1.0:1.4:1.0 for low-voltage-activated ones. 5. The inactivation phase of ICa, HVA was fitted by a sum of double-exponential functions, the time constants of which were larger when the current was carried by Ba2+ than by Ca2+. The inactivation time course of ICa, LVA was fitted by a single-exponential function, and the time constant was practically the same when the current was carried by Ba2+ or by Ca2+. Activation and inactivation processes of ICa, LVA were potential-dependent. 6. The steady-state inactivation curve of ICa, LVA was fitted by the Boltzmann equation, having a mid-potential of -80 mV and a slope factor of 5.0. The recovery time course from steady-state inactivation was fitted by a sum of two exponential functions. The time constants of the faster phase were 230 and 380 ms, and those of slower phase were 2.8 and 1.8 s at the repolarization potentials of -120 and -100 mV, respectively. 7. The amplitude of ICa, LVA depended on the external Ca2+ concentration ([Ca2+]o), approaching saturation at 95 mM [Ca2+]o. 8. Various polyvalent cations blocked both types of Ca2+ current reversibly in the order (IC50 in M): La3+ (8 x 10(-8)) greater than Cd2+ (6 x 10(-6)) greater than Ni2+ (1 x 10(-5)) greater than Zn2+ (2 x 10(-5)) for ICa, HVA, and La3+ (6 x 10(-7)) greater than Zn2+ (3 x 10(-5)) greater than Cd2+ (4 x 10(-4)) greater than Ni2+ (6 x 10(-4)) for ICa, LVA.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 研究了原代培养的大鼠主动脉平滑肌细胞(SMC)中低电压激活的Ca2+电流(ICa,LVA)的电学和药理学特性,特别是与高电压激活的Ca2+电流(ICa,HVA)进行比较。使用全细胞电压钳技术,在含有20 mM - Ca2+的外部溶液中记录这两种类型的Ca2+电流。2. ICa,LVA通过从 - 100 mV的 holding 电位逐步去极化到比 - 60 mV更正的电位来诱发,并在电流 - 电压(I - V)关系中在 - 30 mV左右达到峰值。ICa,HVA在 - 20 mV时被激活,并在 + 20 mV时达到峰值。3. 5 mM - F - 的细胞内透析随时间不可逆地抑制ICa,HVA,而对ICa,LVA影响很小。通过选择 holding 和测试电位水平或通过细胞内灌注F - ,可以将ICa,LVA与ICa,HVA分离。4. 在各自的I - V关系中,高电压激活的Ca2+通道的Ba2+、Sr2+和Ca2+电流的峰值幅度比为1.6:1.2:1.0,低电压激活的通道的该比值为1.0:1.4:1.0。5. ICa,HVA的失活相由双指数函数之和拟合,当电流由Ba2+携带时,其时间常数比由Ca2+携带时更大。ICa,LVA的失活时间进程由单指数函数拟合,当电流由Ba2+或Ca2+携带时,时间常数实际上相同。ICa,LVA的激活和失活过程是电位依赖性的。6. ICa,LVA的稳态失活曲线由玻尔兹曼方程拟合,中点电位为 - 80 mV,斜率因子为5.0。从稳态失活恢复的时间进程由两个指数函数之和拟合。在 - 120和 - 100 mV的复极化电位下,较快相的时间常数分别为230和380 ms,较慢相的时间常数分别为2.8和1.8 s。7. ICa,LVA的幅度取决于外部Ca2+浓度([Ca2+]o),在95 mM [Ca2+]o时接近饱和。8. 各种多价阳离子以(IC50以M计)的顺序可逆地阻断两种类型的Ca2+电流:对于ICa,HVA,La3+(8×10(-8))>Cd2+(6×10(-6))>Ni2+(1×10(-5))>Zn2+(2×10(-5));对于ICa,LVA,La3+(6×10(-7))>Zn2+(3×10(-5))>Cd2+(4×10(-4))>Ni2+(6×10(-4))。(摘要截断于400字)