Lerma J, Zukin R S, Bennett M V
Instituto de Neurobiologia S. Ramón y Cajal, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Proc Natl Acad Sci U S A. 1990 Mar;87(6):2354-8. doi: 10.1073/pnas.87.6.2354.
In Xenopus oocytes injected with rat brain mRNA, as in neurons, glycine greatly potentiated responses of the N-methyl-D-aspartate (NMDA) type of excitatory amino acid receptor. Injected oocytes generated a partially desensitizing inward current in response to NMDA with 30 nM added glycine. As the added glycine concentration was increased from 30 nM to 1 microM, the NMDA response was increased and exhibited less desensitization. The relationship between the NMDA peak response and added glycine concentration indicated a single component response with apparent affinity of 0.29 microM and a Hill coefficient of 0.77. The desensitized response was also fit by the Hill relation with a lower affinity but similar coefficient. The time course of desensitization at 500 microM NMDA was exponential with a time constant (350 msec) that was independent of glycine concentration between 0.03 and 0.3 microM. At higher glycine concentration a slower component of decay (tau = 1.4 sec) was observed. This component was enhanced by increasing the extracellular Ca2+. NMDA without added glycine evoked a small transient response. However this response was suppressed completely by prewashing with the glycine antagonist 7-chlorokynurenic acid, suggesting that it may have been due to glycine contamination. The dose-response relation for low concentrations of glycine indicated that the measured level of glycine contamination accounted for these responses. These results indicate that glycine has at least two actions at the NMDA receptor: it enables channel opening by the agonist and decreases desensitization.
在注射了大鼠脑信使核糖核酸(mRNA)的非洲爪蟾卵母细胞中,与神经元一样,甘氨酸极大地增强了N-甲基-D-天冬氨酸(NMDA)型兴奋性氨基酸受体的反应。注射的卵母细胞在添加30 nM甘氨酸的情况下,对NMDA产生部分脱敏的内向电流。随着添加的甘氨酸浓度从30 nM增加到1 μM,NMDA反应增强且脱敏程度降低。NMDA峰值反应与添加的甘氨酸浓度之间的关系表明存在单一组分反应,表观亲和力为0.29 μM,希尔系数为0.77。脱敏反应也符合希尔关系,亲和力较低但系数相似。在500 μM NMDA时脱敏的时间进程呈指数关系,时间常数为350毫秒,在0.03至0.3 μM的甘氨酸浓度范围内与甘氨酸浓度无关。在较高的甘氨酸浓度下,观察到较慢的衰减成分(时间常数τ = 1.4秒)。增加细胞外钙离子(Ca2+)可增强该成分。未添加甘氨酸的NMDA引起小的瞬时反应。然而,用甘氨酸拮抗剂7-氯犬尿氨酸预洗可完全抑制该反应,表明这可能是由于甘氨酸污染所致。低浓度甘氨酸的剂量反应关系表明,测得的甘氨酸污染水平可解释这些反应。这些结果表明,甘氨酸在NMDA受体上至少有两种作用:它通过激动剂使通道开放并减少脱敏。