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大鼠急性分离的大细胞胆碱能基底前脑神经元中的全细胞钙电流

The whole-cell calcium current in acutely dissociated magnocellular cholinergic basal forebrain neurones of the rat.

作者信息

Allen T G, Sim J A, Brown D A

机构信息

Department of Pharmacology, University College London.

出版信息

J Physiol. 1993 Jan;460:91-116. doi: 10.1113/jphysiol.1993.sp019461.

Abstract
  1. The electrophysiological and pharmacological characteristics of the calcium current (ICa) in acutely dissociated magnocellular cholinergic basal forebrain neurones from 11- to 14-day-old post-natal rats were studied using the whole-cell patch-clamp technique. 2. All cells exhibited a small transient low-voltage-activated (LVA) current with half-activation and half-inactivation potentials of -40.2 and -49.3 mV and slope factors for activation and inactivation of 4.82 and 3.85 mV per e-fold change in membrane potential (Vm) respectively. Activation and inactivation rates for the LVA current were highly voltage dependent. For test potential changes from -50 to -20 mV, the time-to-peak of the current decreased from 39.1 to 6.4 ms, and the time constant of current decay decreased from 81.7 to 15.5 ms. 3. A high-voltage-activated (HVA) component of ICa could be elicited at threshold voltages between -46 and -30 mV from a holding potential (VH) of -80 mV. The HVA current peaked around 0 mV; a 10-fold increase in [Ca2+]o produced a 13 mV positive shift in the peak, whilst the amplitude of the current showed an approximately hyperbolic relationship to [Ca2+]o with half-saturation at 2.5 mM. The transient phase of the HVA current could be described by two exponential functions with time constants tau fast and tau slow of 16.2 and 301 ms. Steady-state inactivation of the transient and extrapolated true sustained (pedestal) components of HVA current were described by Boltzmann equations, with half-inactivation potentials (slope factors) of -47.3 mV, (9.04) and -29.2 mV (11.8) respectively. 4. omega-Conotoxin (omega-CgTX; 100 nM) irreversibly inhibited a kinetically distinct component of HVA current but had no effect upon the transient LVA current. The omega-CgTX-sensitive current could not be distinguished from the control HVA current by the voltage dependence of its activation or inactivation rates. 5. Low concentrations of amiloride (< or = 300 microM) or Ni2+ (< or = 5 microM) selectively inhibited the transient LVA current, with IC50 values of 97 and 5 microM respectively. Cd2+ (< or = 1 microM) selectively blocked a component of HVA current. At higher concentrations, Cd2+ and Ni2+ were non-selective and totally blocked all components of ICa. 6. The lanthanide ions Gd3+ and La3+ produced saturable incomplete block of the HVA current. Maximally effective concentrations of Gd3+ (100 microM) or La3+ (30 microM) inhibited 76.5 and 41.2% respectively of the sustained component of HVA current with corresponding IC50 values of 2.2 and 1.1 microM.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 采用全细胞膜片钳技术,研究了出生后11至14天大鼠急性分离的大细胞胆碱能基底前脑神经元钙电流(ICa)的电生理和药理学特性。2. 所有细胞均表现出一种小的瞬时低电压激活(LVA)电流,其半激活和半失活电位分别为-40.2和-49.3 mV,激活和失活的斜率因子分别为每膜电位(Vm)变化1倍时4.82和3.85 mV。LVA电流的激活和失活速率高度依赖电压。对于测试电位从-50 mV变化到-20 mV,电流的峰值时间从39.1 ms降至6.4 ms,电流衰减的时间常数从81.7 ms降至15.5 ms。3. 从-80 mV的钳制电位(VH)开始,在-46至-30 mV的阈值电压下可诱发ICa的高电压激活(HVA)成分。HVA电流在0 mV左右达到峰值;细胞外钙离子浓度([Ca2+]o)增加10倍,峰值正向偏移13 mV,而电流幅度与[Ca2+]o呈现近似双曲线关系,半饱和浓度为2.5 mM。HVA电流的瞬时相可用两个指数函数描述,快速时间常数(tau fast)和慢速时间常数(tau slow)分别为16.2和301 ms。HVA电流的瞬时成分和外推的真正持续(基座)成分的稳态失活可用玻尔兹曼方程描述,半失活电位(斜率因子)分别为-47.3 mV(9.04)和-29.2 mV(11.8)。4. ω-芋螺毒素(ω-CgTX;100 nM)不可逆地抑制HVA电流中动力学上不同的一个成分,但对瞬时LVA电流无影响。ω-CgTX敏感电流在激活或失活速率的电压依赖性方面与对照HVA电流无法区分。5. 低浓度的氨氯地平(≤300 μM)或镍离子(≤5 μM)选择性抑制瞬时LVA电流,IC50值分别为97和5 μM。镉离子(≤1 μM)选择性阻断HVA电流的一个成分。在较高浓度时,镉离子和镍离子无选择性,完全阻断ICa的所有成分。6. 镧系离子钆离子(Gd3+)和镧离子(La3+)对HVA电流产生饱和性不完全阻断。Gd3+(100 μM)或La3+(30 μM)的最大有效浓度分别抑制HVA电流持续成分的76.5%和41.2%,相应的IC50值分别为2.2和1.1 μM。(摘要截短至400字)

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