Nabekura Junichi, Ueno Tsuyoshi, Katsurabayashi Shutaro, Furuta Akiko, Akaike Norio, Okada Masayoshi
Department of Cellular and System Physiology, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka 812-8582, Japan.
J Physiol. 2002 Mar 15;539(Pt 3):735-41. doi: 10.1113/jphysiol.2001.013379.
To elucidate characteristic changes in the N-methyl-D-aspartate (NMDA) receptor on neurons following axotomy, subunit expressions and functional features of the NMDA receptor were examined in the dorsal motor nucleus of vagus (DMV) of rats receiving vagal axotomy at the neck. Western blotting analysis demonstrated that the expression of NR2A decreased 2-3 days after in vivo axotomy, while expression of NR1 and NR2B, NR2C and NR2D subunits did not change significantly. To examine the functional changes, patch clamp recordings in whole-cell mode were employed on the axotomized DMV neurons identified by retrograde labelling with fluorescent dye. The amplitude ratios of ifenprodil-sensitive components of NMDA response and D,L-2-amino-5-phosphovaleric acid (APV)-sensitive evoked postsynaptic current increased after axotomy. In addition, APV-sensitive postsynaptic currents exhibited a longer decay time in identified axotomized vagal motoneurons than in control neurons. No significant differences in the current density of the NMDA response and the peak amplitude of APV-sensitive synaptic currents were observed between axotomized and intact DMV neurons. In conclusion, a decrease in NR2A expression results in the appearance of functional characteristics of the NMDA receptor predominantly containing the NR2B subunit. This might lead to a long-term increase of the susceptibility of neurons to excitotoxicity.
为阐明轴突切断术后神经元上N-甲基-D-天冬氨酸(NMDA)受体的特征性变化,在颈部接受迷走神经切断术的大鼠迷走神经背核(DMV)中检测了NMDA受体的亚基表达和功能特性。蛋白质免疫印迹分析表明,体内轴突切断术后2-3天,NR2A的表达下降,而NR1、NR2B、NR2C和NR2D亚基的表达没有明显变化。为检测功能变化,采用全细胞膜片钳记录法,对经荧光染料逆行标记鉴定的轴突切断的DMV神经元进行记录。轴突切断术后,NMDA反应中ifenprodil敏感成分与D,L-2-氨基-5-磷酸缬氨酸(APV)敏感的诱发突触后电流的幅度比增加。此外,在已鉴定的轴突切断的迷走运动神经元中,APV敏感的突触后电流的衰减时间比对照神经元更长。轴突切断的和完整的DMV神经元之间,在NMDA反应的电流密度和APV敏感的突触电流的峰值幅度方面未观察到显著差异。总之,NR2A表达的降低导致主要包含NR2B亚基的NMDA受体功能特性的出现。这可能导致神经元对兴奋性毒性的易感性长期增加。