Gruol D L, Jacquin T, Yool A J
Department of Neuropharmacology, Research Institute of Scripps Clinic, La Jolla, California 92037.
J Neurosci. 1991 Apr;11(4):1002-15. doi: 10.1523/JNEUROSCI.11-04-01002.1991.
Voltage-sensitive K+ channels were studied in rat cerebellar Purkinje neurons in culture using the single-channel recording technique. Recordings in the cell-attached and outside-out configuration revealed multiple voltage-sensitive K+ channel types in patches from both the somatic and the dendritic regions. K+ channel types were present in all patches studied. The same channel types were observed in somatic and dendritic recordings. Channel types were identified by reversal potential, single-channel conductance, voltage sensitivity, and patterns of activity. In cell-attached patches recorded under physiological conditions, 3 channel types were identified. Mean single-channel conductances were 92, 57, and 12 pS. All 3 channel types were activated by membrane depolarization. Similar channel types were identified in inside-out and outside-out patches recorded under physiological conditions. Two additional channel types were identified in the outside-out patches, with mean single-channel conductances of 41 and 26 pS. In cell-attached recordings under symmetrical K+ conditions, 6 channel types were identified. Mean single-channel conductances were 222, 134, 39, 25, 14, and 15 pS. Channel types with mean conductances of 222, 134, and 39 pS required membrane depolarization for activation. A comparison of channel properties indicated that these channel types correlated with the 3 channel types observed in cell-attached patches under physiological conditions. The 3 smaller-conductance channel types (25, 14, and 15 pS) were active at potentials around rest or at hyperpolarized membrane potentials. Two K+ channel types (39 and 25 pS) were commonly associated with the late phase of extracellularly recorded spontaneous spike events, suggesting a functional role in the repolarizing phase of somatic and dendritic action potentials. These results demonstrate that voltage-sensitive K+ channels are a prominent component of both the somatic and the dendritic membrane of the cerebellar Purkinje neuron and support the view that multiple voltage-sensitive K+ channel types contribute to the membrane functions of both cellular regions in this CNS neuronal type.
运用单通道记录技术,对培养的大鼠小脑浦肯野神经元中的电压敏感性钾通道进行了研究。在细胞贴附式和膜外翻式配置下的记录显示,在体区和树突区的膜片中存在多种电压敏感性钾通道类型。在所研究的所有膜片中均存在钾通道类型。在体记录和树突记录中观察到相同的通道类型。通过反转电位、单通道电导、电压敏感性和活动模式来识别通道类型。在生理条件下记录的细胞贴附式膜片中,识别出了3种通道类型。平均单通道电导分别为92、57和12 pS。所有这3种通道类型均由膜去极化激活。在生理条件下记录的膜外翻式和内面向外式膜片中也识别出了类似的通道类型。在内面向外式膜片中还识别出另外2种通道类型,平均单通道电导分别为41和26 pS。在对称钾条件下的细胞贴附式记录中,识别出了6种通道类型。平均单通道电导分别为222、134、39、25、14和15 pS。平均电导为222、134和39 pS的通道类型需要膜去极化才能激活。通道特性的比较表明,这些通道类型与在生理条件下细胞贴附式膜片中观察到的3种通道类型相关。3种较小电导的通道类型(25、14和15 pS)在静息电位附近或超极化膜电位时处于激活状态。2种钾通道类型(39和25 pS)通常与细胞外记录的自发尖峰事件的后期相关,表明在体和树突动作电位的复极化阶段具有功能作用。这些结果表明,电压敏感性钾通道是小脑浦肯野神经元体膜和树突膜的重要组成部分,并支持这样一种观点,即多种电压敏感性钾通道类型对这种中枢神经系统神经元类型的两个细胞区域的膜功能都有贡献。