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关于一种人类神经母细胞瘤细胞系的钠电流和钾电流

On the sodium and potassium currents of a human neuroblastoma cell line.

作者信息

Ginsborg B L, Martin R J, Patmore L

机构信息

Department of Pharmacology, University of Edinburgh.

出版信息

J Physiol. 1991 Mar;434:121-49. doi: 10.1113/jphysiol.1991.sp018462.

Abstract
  1. The patch-clamp method was applied to the study of ionic currents activated by depolarization of undifferentiated IMR-32 human neuroblastoma cells. Whole-cell sodium and potassium currents and single potassium ion channel currents from cell-attached patches were investigated. 2. Cells had a mean resting potential of -38 mV and mean input resistance of 1.6 G omega. Single action potentials were evoked under current clamp during the injection of depolarizing currents. 3. A voltage-dependent inward sodium current was observed which reversed at +44 mV. A Boltzmann fit to the activation curve gave a half-maximal activation voltage of -41.6 mV and a 'slope' of 3.9 mV. The steady-state inactivation curve had a half-maximal inactivation voltage of -81 mV and a 'slope' of 9.7 mV. 4. The time-dependent activation and inactivation of the current displayed classical Hodgkin-Huxley kinetics. Values for the time constants tau m and tau h of 0.16 and 0.63 ms were calculated for a voltage jump from -80 to -10 mV; tau m and tau h decreased as the step potential was changed from -30 to +20 mV. 5. Outward currents were activated in bathing solutions substantially free of anions and could thus be attributed to potassium ions. The tail current reversed in direction on repolarization to -60 mV when the potassium concentration in the bathing solution was increased from 6 to 30 mM. When the bathing solution contained 145 mM-potassium, and the patch pipette, 95 mM, a depolarization to -10 mV from a holding potential of -60 mV evoked an inward current. 6. Outward currents were examined by using voltage pulses which depolarized the cell to -20 mV, or more positive values, from a holding potential of -80 mV and by pulses which depolarized the cell to 0 mV, or to positive values, from a holding potential of -30 mV. A Boltzmann fit of typical activation data gave a half-maximal activation voltage of 17 mV and a 'slope' of 14 mV. 7. The time course of the rising phase of the current was described by a function of the form A(1-exp[-(t-delta t)/tau]), where delta t varied between 1 and 4 ms and tau varied between 4 and 27 ms, decreasing with increasing depolarization. There was no evidence for a fast transient component. 8. The amplitude of outward currents was reduced by extracellular calcium ions, cobalt ions, tetraethylammonium and 4-aminopyridine.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 膜片钳技术被用于研究未分化的人IMR - 32神经母细胞瘤细胞去极化激活的离子电流。研究了全细胞钠电流和钾电流以及细胞贴附式膜片的单个钾离子通道电流。2. 细胞的平均静息电位为 - 38 mV,平均输入电阻为1.6 GΩ。在电流钳模式下注入去极化电流时可诱发单个动作电位。3. 观察到一种电压依赖性内向钠电流,其反转电位为 + 44 mV。对激活曲线进行玻尔兹曼拟合得到半最大激活电压为 - 41.6 mV,“斜率”为3.9 mV。稳态失活曲线的半最大失活电压为 - 81 mV,“斜率”为9.7 mV。4. 电流的时间依赖性激活和失活表现出经典的霍奇金 - 赫胥黎动力学。对于从 - 80 mV到 - 10 mV的电压阶跃,计算得到时间常数τm和τh的值分别为0.16和0.63 ms;当阶跃电位从 - 30 mV变为 + 20 mV时,τm和τh减小。5. 在基本无阴离子的浴液中可激活外向电流,因此可归因于钾离子。当浴液中的钾浓度从6 mM增加到30 mM时,复极化到 - 60 mV时尾电流方向反转。当浴液含有145 mM - 钾,膜片微电极含有95 mM钾时,从 - 60 mV的钳制电位去极化到 - 10 mV可诱发内向电流。6. 通过使用从 - 80 mV的钳制电位将细胞去极化到 - 20 mV或更正电位的电压脉冲,以及从 - 30 mV的钳制电位将细胞去极化到0 mV或正电位的脉冲来检测外向电流。对典型激活数据进行玻尔兹曼拟合得到半最大激活电压为17 mV,“斜率”为14 mV。7. 电流上升相的时间进程可用A(1 - exp[ - (t - Δt)/τ])形式的函数描述,其中Δt在1到4 ms之间变化,τ在4到27 ms之间变化,随去极化增加而减小。没有证据表明存在快速瞬态成分。8. 细胞外钙离子、钴离子、四乙铵和4 - 氨基吡啶可降低外向电流的幅度。(摘要截断于400字)

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