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内侧前庭核神经元中离子型谷氨酸受体的电生理和药理学特性:急性分离神经元的全细胞膜片钳研究

Electrophysiological and pharmacological characteristics of ionotropic glutamate receptors in medial vestibular nucleus neurons: a whole cell patch clamp study in acutely dissociated neurons.

作者信息

Sakai N, Ujihara H, Ishihara K, Sasa M, Tanaka C

机构信息

Laboratory of Molecular Pharmacology, Kobe University, Japan.

出版信息

Jpn J Pharmacol. 1996 Dec;72(4):335-46. doi: 10.1254/jjp.72.335.

Abstract

A patch clamp study was performed to determine which subtype of ionotropic glutamate receptors is involved in the glutamate-induced excitation of the medial vestibular nucleus (MVN) neurons. Whole cell recording was performed on MVN neurons that were acutely dissociated by enzymatic and mechanical treatments. Application of glutamate at a concentration of 100 microM produced a current with a reversal potential of approximately 0 mV. The glutamate-induced current was completely blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), a non-N-methyl-D-aspartate (NMDA)-receptor antagonist. Application of alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) and kainic acid (KA), non-NMDA-receptor agonists, at concentrations of 30 and 100 microM produced a concentration-dependent depolarization concomitantly with an increase in firing rates during current clamp recording. During voltage clamp recording, glutamate, AMPA and KA elicited a concentration-dependent current with an equilibrium potential of approximately 0 mV. To clarify whether NMDA receptors are present in MVN neurons, the effects of glycine on the glutamate- and NMDA-induced current were examined. Two types of NMDA receptor-mediated current (types 1 and 2) were obtained in terms of the difference in sensitivity to both magnesium ion and MK-801, which act on the NMDA-receptor channel. In the type 1 neurons, the NMDA-induced current was not apparently blocked by magnesium ion or MK-801, although a larger current was obtained in the absence of magnesium ion. In the type 2 neurons, marked blockade of the NMDA-induced current was seen in the presence of magnesium ion and MK-801, as previously reported in other neurons of the central nervous system. These findings indicate the presence of both non-NMDA and NMDA receptors, which are involved in primary afferent transmission, in the MVN neuron, and two distinct types of NMDA receptors.

摘要

进行了膜片钳研究,以确定离子型谷氨酸受体的哪种亚型参与谷氨酸诱导的内侧前庭核(MVN)神经元兴奋。对经酶解和机械处理急性解离的MVN神经元进行全细胞记录。施加浓度为100微摩尔的谷氨酸产生了反转电位约为0毫伏的电流。谷氨酸诱导的电流被非N-甲基-D-天冬氨酸(NMDA)受体拮抗剂6-氰基-7-硝基喹喔啉-2,3-二酮(CNQX,10微摩尔)完全阻断。施加浓度为30和100微摩尔的非NMDA受体激动剂α-氨基-3-羟基-5-甲基异恶唑-4-丙酸(AMPA)和海人酸(KA),在电流钳记录期间产生浓度依赖性去极化,同时放电频率增加。在电压钳记录期间,谷氨酸、AMPA和KA引发了平衡电位约为0毫伏的浓度依赖性电流。为了阐明MVN神经元中是否存在NMDA受体,研究了甘氨酸对谷氨酸和NMDA诱导电流的影响。根据对作用于NMDA受体通道的镁离子和MK-801的敏感性差异,获得了两种类型的NMDA受体介导电流(1型和2型)。在1型神经元中,NMDA诱导的电流虽然在无镁离子时获得的电流更大,但并未明显被镁离子或MK-801阻断。在2型神经元中,如先前在中枢神经系统的其他神经元中所报道的,在存在镁离子和MK-801的情况下,NMDA诱导的电流明显被阻断。这些发现表明MVN神经元中存在参与初级传入传递的非NMDA和NMDA受体,以及两种不同类型的NMDA受体。

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