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急性分离的大鼠新纹状体神经元中的多种钙通道类型。

Multiple types of calcium channels in acutely isolated rat neostriatal neurons.

作者信息

Hoehn K, Watson T W, MacVicar B A

机构信息

Neuroscience Research Group, University of Calgary, Alberta, Canada.

出版信息

J Neurosci. 1993 Mar;13(3):1244-57. doi: 10.1523/JNEUROSCI.13-03-01244.1993.

Abstract

Voltage-activated high- and low-threshold Ca2+ currents were studied using whole-cell voltage-clamp techniques and fura-2 fluorescence measurements of intracellular Ca2+ in neurons acutely isolated from rat neostriatum. High-threshold Ca2+ currents activated around -40 mV and were present in at least 95% of neostriatal neurons. The maximum current, 736 +/- 44 pA (mean +/- SEM, n = 141), was observed around 0 mV. In 70% of neurons, high-threshold Ca2+ currents exhibited both inactivating and noninactivating components. The majority of the high-threshold Ca2+ currents appeared to belong neither to the "L-type" nor the "N-type" classification, since omega-conotoxin (5 microM) decreased this current by only 29% and nimodipine (10 microM) decreased the noninactivating component of this current by only 17%. A low-threshold transient (T-type) Ca2+ current was observed in 40% of neurons. When both T-type and high-threshold Ca2+ currents were present, their maximum amplitudes were 78 +/- 7 pA and 800 +/- 57 pA, respectively (mean +/- SEM, n = 58). At a holding potential of -100 mV, the T-type Ca2+ current activated around -60 mV, with maximum current near -40 mV. Steady-state inactivation of the T-type Ca2+ current was observed at holding potentials positive to -125 mV, and the current was half-inactivated at -88 mV. Recovery from inactivation to 90% of maximum occurred within 800 msec. Mn2+ or Co2+ (3 mM) blocked both high-threshold and T-type Ca2+ currents, whereas Cd2+ (25 microM) or verapamil (50 microM and 150 microM) preferentially blocked high-threshold over T-type Ca2+ currents. In response to depolarization by 50 mM K+, fura-2 fluorescence measurements showed increased intracellular Ca2+ in both the soma and the proximal dendrites of neostriatal neurons that was markedly reduced by 25 microM Cd2+. These findings suggest that high-threshold Ca channels are present in both the soma and proximal dendrites of neostriatal neurons.

摘要

采用全细胞膜片钳技术以及对急性分离自大鼠新纹状体的神经元进行fura-2荧光测量细胞内Ca2+的方法,研究了电压门控的高阈值和低阈值Ca2+电流。高阈值Ca2+电流在约-40 mV时激活,至少95%的新纹状体神经元中存在该电流。最大电流为736±44 pA(平均值±标准误,n = 141),在约0 mV时观察到。在70%的神经元中,高阈值Ca2+电流表现出失活和非失活成分。大多数高阈值Ca2+电流似乎既不属于“L型”也不属于“N型”分类,因为ω-芋螺毒素(5 μM)仅使该电流降低29%,尼莫地平(10 μM)仅使该电流的非失活成分降低17%。在40%的神经元中观察到低阈值瞬态(T型)Ca2+电流。当T型和高阈值Ca2+电流同时存在时,它们的最大幅度分别为78±7 pA和800±57 pA(平均值±标准误,n = 58)。在-100 mV的钳制电位下,T型Ca2+电流在约-60 mV时激活,最大电流接近-40 mV。在高于-125 mV的钳制电位下观察到T型Ca2+电流的稳态失活,该电流在-88 mV时半失活。从失活恢复到最大电流的90%在800毫秒内发生。3 mM的Mn2+或Co2+可阻断高阈值和T型Ca2+电流,而25 μM的Cd2+或维拉帕米(50 μM和150 μM)优先阻断高阈值Ca2+电流而非T型Ca2+电流。响应50 mM K+引起的去极化,fura-2荧光测量显示新纹状体神经元的胞体和近端树突内的细胞内Ca2+增加,25 μM Cd2+可使其显著降低。这些发现表明高阈值Ca通道存在于新纹状体神经元的胞体和近端树突中。

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