Williams K
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia 19104-6084.
Mol Pharmacol. 1993 Oct;44(4):851-9.
The effects of the atypical N-methyl-D-aspartate (NMDA) receptor antagonist ifenprodil were investigated by voltage-clamp recording of Xenopus oocytes expressing heteromeric NMDA receptors from cloned NR1 and NR2 subunit RNAs. In oocytes voltage-clamped at -70 mV, ifenprodil inhibited NMDA-induced currents at NR1A/NR2B receptors with high affinity (IC50 = 0.34 microM). The affinity of NR1A/NR2A receptors for ifenprodil (IC50 = 146 microM) was 400-fold lower than that of NR1A/NR2B receptors. The rate of onset of inhibition by low concentrations of ifenprodil acting at NR1A/NR2B receptors was considerably slower than the onset of inhibition seen with high concentrations of ifenprodil acting at NR1A/NR2A receptors. The onset and recovery of blockade by ifenprodil at NR1A/NR2B receptors were not activity dependent. The inhibitory effects of low concentrations of ifenprodil at NR1A/NR2B receptors were not voltage dependent. In contrast, the inhibitory effects of high concentrations of ifenprodil at NR1A/NR2A receptors were partially voltage dependent, and a greater inhibition of NMDA-induced currents was seen at hyperpolarized membrane potentials than at depolarized membrane potentials. The reversal potential of NMDA currents was not altered in the presence of ifenprodil. Ifenprodil may act as a weak open-channel blocker of NR1A/NR2A receptors. The degree of inhibition seen with 100 microM ifenprodil at NR1A/NR2A receptors was not altered by changes in the concentration of extracellular glycine. However, the inhibitory effect of 1 microM ifenprodil at NR1A/NR2B receptors was reduced by increasing the concentration of glycine. Thus, part of the mechanism of action of ifenprodil at NR1A/NR2B receptors may involve noncompetitive antagonism of the effects of glycine. These results indicate that the mechanism of action of ifenprodil, as well as the potency of this antagonist, is different at NR1A/NR2B and NR1A/NR2A receptors expressed in Xenopus oocytes.
通过电压钳记录非洲爪蟾卵母细胞中由克隆的NR1和NR2亚基RNA表达的异聚N-甲基-D-天冬氨酸(NMDA)受体,研究了非典型NMDA受体拮抗剂艾芬地尔的作用。在钳制电压为-70 mV的卵母细胞中,艾芬地尔以高亲和力抑制NR1A/NR2B受体处NMDA诱导的电流(IC50 = 0.34 microM)。NR1A/NR2A受体对艾芬地尔的亲和力(IC50 = 146 microM)比对NR1A/NR2B受体的亲和力低400倍。低浓度艾芬地尔作用于NR1A/NR2B受体时的抑制起效速率明显慢于高浓度艾芬地尔作用于NR1A/NR2A受体时的抑制起效速率。艾芬地尔对NR1A/NR2B受体的阻断起效和恢复不依赖于活性。低浓度艾芬地尔对NR1A/NR2B受体的抑制作用不依赖于电压。相反,高浓度艾芬地尔对NR1A/NR2A受体的抑制作用部分依赖于电压,与去极化膜电位相比,在超极化膜电位下对NMDA诱导电流的抑制作用更强。在存在艾芬地尔的情况下,NMDA电流的反转电位未改变。艾芬地尔可能作为NR1A/NR2A受体的弱开放通道阻滞剂。细胞外甘氨酸浓度的变化未改变100 microM艾芬地尔对NR1A/NR2A受体的抑制程度。然而,增加甘氨酸浓度可降低1 microM艾芬地尔对NR1A/NR2B受体的抑制作用。因此,艾芬地尔在NR1A/NR2B受体处的部分作用机制可能涉及对甘氨酸作用的非竞争性拮抗。这些结果表明,艾芬地尔在非洲爪蟾卵母细胞中表达的NR1A/NR2B和NR1A/NR2A受体上的作用机制以及该拮抗剂的效力是不同的。