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人Jurkat T细胞系中Ca2+和K+电流的特征:植物血凝素的作用

Characterization of Ca2+ and K+ currents in the human Jurkat T cell line: effects of phytohaemagglutinin.

作者信息

Dupuis G, Héroux J, Payet M D

机构信息

Department of Biochemistry, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.

出版信息

J Physiol. 1989 May;412:135-54. doi: 10.1113/jphysiol.1989.sp017608.

DOI:10.1113/jphysiol.1989.sp017608
PMID:2557424
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1190568/
Abstract
  1. Inward and outward currents were recorded in the human Jurkat T cell line using the whole-cell configuration of the patch-clamp technique. 2. The transient outward current was activated at membrane potentials positive to -60 mV. The activation time constant-voltage relationship decreased from 17 ms to 2 ms for membrane potentials ranging from -40 to +40 mV. The inactivation phase could be fitted by a single-exponential function and the inactivation time constant decreased from 250 ms to 150 ms for membrane potentials ranging from -20 to +100 mV. 3. The steady-state inactivation-voltage relationship showed a mid-point potential of -32 +/- 2.6 mV, and the slope factor was 10.8 +/- 1.8 mv (n = 3). 4. The calcium ionophore A23187 provoked a decrease in the amplitude of the outward current, suggesting a dependence of this current on the cytosolic concentration of Ca2+. 5. The K+ outward current was blocked by tetraethylammonium (TEA, Michaelis-Menten constant (Km), 6 mM) and by the calcium channel blockers Ni2+, Co2+, Mn2+ and Cd2+. 6. Forty per cent (n = 120) of the patched Jurkat cells displayed an inward current. In a physiological medium containing Ca2+ (2.2 mM), the inward current threshold voltage was -60 mV, the maximum current was observed at -40 mV and the zero current voltage was positive to +20 mV. At negative membrane potentials, the time required to reach 50% of the maximum amplitude was 60 ms and grew shorter with increasing depolarization, reaching a value of 5 ms at -5 mV. The inactivation of the inward current was very slow and the time constant varied from 1200 ms at -35 mV to approximately 250 ms for potentials positive to -10 mV. 7. The current availability had a value of one for potentials negative to -50 mV and zero for potentials positive to -15 mV. The mid-point potential was -31 +/- 3.4 mV and the slope factor was 3.3 +/- 0.2 mV (n = 3). 8. The inward channels were permeable to Sr2+, but were blocked by classical Ca2+ channel inhibitors such as Co2+, Mn2+ and Ni2+. 9. Phaseolus vulgaris phytohaemagglutinin (PHA), an inducer of interleukin-2 production in Jurkat cells, increased the inward current amplitude by 32 +/- 20% (n = 4). This increase was concomitant with a decrease (45 +/- 12%) in the amplitude of the outward current, but only when the current was carried by Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 采用膜片钳技术的全细胞模式记录人Jurkat T细胞系的内向和外向电流。2. 瞬时外向电流在膜电位高于 -60 mV时被激活。对于膜电位从 -40 mV到 +40 mV的范围,激活时间常数 - 电压关系从17 ms降至2 ms。失活相可用单指数函数拟合,对于膜电位从 -20 mV到 +100 mV的范围,失活时间常数从250 ms降至150 ms。3. 稳态失活 - 电压关系显示中点电位为 -32±2.6 mV,斜率因子为10.8±1.8 mV(n = 3)。4. 钙离子载体A23187引起外向电流幅度减小,表明该电流依赖于胞质Ca2+浓度。5. K+外向电流被四乙铵(TEA,米氏常数(Km),6 mM)以及钙通道阻滞剂Ni2+、Co2+、Mn2+和Cd2+阻断。6. 40%(n = 120)的膜片钳Jurkat细胞显示有内向电流。在含有Ca2+(2.2 mM)的生理介质中,内向电流阈值电压为 -60 mV,在 -40 mV时观察到最大电流,零电流电压高于 +20 mV。在负膜电位下,达到最大幅度50%所需时间为60 ms,且随着去极化增加而缩短,在 -5 mV时达到5 ms。内向电流的失活非常缓慢,时间常数从 -35 mV时的1200 ms变化到高于 -10 mV电位时的约250 ms。7. 对于低于 -50 mV的电位,电流可用性值为1,对于高于 -15 mV的电位,电流可用性值为零。中点电位为 -31±3.4 mV,斜率因子为3.3±0.2 mV(n = 3)。8. 内向通道对Sr2+通透,但被经典的Ca2+通道抑制剂如Co2+、Mn2+和Ni2+阻断。9. 菜豆植物血凝素(PHA)是Jurkat细胞中白细胞介素 -2产生的诱导剂,使内向电流幅度增加32±20%(n = 4)。这种增加伴随着外向电流幅度的减小(45±12%),但仅当电流由Ca2+携带时。(摘要截短于400字)

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