Kemp J A, Foster A C, Leeson P D, Priestley T, Tridgett R, Iversen L L, Woodruff G N
Merck Sharp & Dohme Research Laboratories, Neuroscience Research Centre, Harlow, Essex, United Kingdom.
Proc Natl Acad Sci U S A. 1988 Sep;85(17):6547-50. doi: 10.1073/pnas.85.17.6547.
Glycine markedly potentiates N-methyl-D-aspartate (N-Me-D-Asp) responses in mammalian neurons by an action at a modulatory site on the N-Me-D-Asp receptor-ionophore complex. Here we present evidence that 7-chlorokynurenic acid (7-Cl KYNA) inhibits N-Me-D-Asp responses by a selective antagonism of glycine at this modulatory site. In rat cortical slices 7-Cl KYNA (10-100 microM) noncompetitively inhibited N-Me-D-Asp responses, and this effect could be reversed by the addition of glycine (100 microM) or D-serine (100 microM). Radioligand binding experiments showed that 7-Cl KYNA had a much higher affinity for the strychnine-insensitive [3H]glycine binding site (IC50 = 0.56 microM) than for the N-Me-D-Asp (IC50 169 microM), quisqualate (IC50 = 153 microM), or kainate (IC50 greater than 1000 microM) recognition sites. In whole-cell patch-clamp recordings from rat cortical neurones in culture, the inhibitory effects of 7-Cl KYNA on N-Me-D-Asp-induced currents could not be overcome by increasing the N-Me-D-Asp concentration but could be reversed by increasing the glycine concentration. 7-Cl KYNA could completely abolish N-Me-D-Asp responses, including basal responses in the absence of added glycine, suggesting that it may possess negative modulatory effects at the glycine site. These findings indicate that the glycine modulatory site is functional in intact adult tissue and that 7-Cl KYNA should prove to be a selective tool for elucidating the involvement of this site in physiological and pathological events mediated by N-Me-D-Asp receptors.
甘氨酸通过作用于N-甲基-D-天冬氨酸(N-Me-D-Asp)受体-离子载体复合物上的一个调节位点,显著增强哺乳动物神经元对N-Me-D-Asp的反应。在此,我们提供证据表明,7-氯犬尿酸(7-Cl KYNA)通过在该调节位点选择性拮抗甘氨酸来抑制N-Me-D-Asp反应。在大鼠皮质切片中,7-Cl KYNA(10 - 100微摩尔)非竞争性抑制N-Me-D-Asp反应,添加甘氨酸(100微摩尔)或D-丝氨酸(100微摩尔)可逆转此效应。放射性配体结合实验表明,7-Cl KYNA对士的宁不敏感的[3H]甘氨酸结合位点(IC50 = 0.56微摩尔)的亲和力比对N-Me-D-Asp(IC50 = 169微摩尔)、quisqualate(IC50 = 153微摩尔)或海人藻酸(IC50大于1000微摩尔)识别位点的亲和力高得多。在培养的大鼠皮质神经元的全细胞膜片钳记录中,增加N-Me-D-Asp浓度无法克服7-Cl KYNA对N-Me-D-Asp诱导电流的抑制作用,但增加甘氨酸浓度可逆转该抑制作用。7-Cl KYNA可完全消除N-Me-D-Asp反应,包括在不添加甘氨酸时的基础反应,这表明它可能在甘氨酸位点具有负性调节作用。这些发现表明,甘氨酸调节位点在完整的成年组织中具有功能,并且7-Cl KYNA应被证明是阐明该位点在由N-Me-D-Asp受体介导的生理和病理事件中所起作用的一种选择性工具。