Differential sensitivity of recombinant N-methyl-D-aspartate receptor subtypes to zinc inhibition

Mol Pharmacol. 1997 Jun;51(6):1015-23. doi: 10.1124/mol.51.6.1015.

Abstract

Zinc has been shown to be present in synaptic vesicles of a subset of glutamatergic boutons and is believed to be core-leased with glutamate at these synapses. A variety of studies have suggested that zinc might play a role in modulation of excitatory transmission, as well as excitotoxicity, by inhibiting N-methyl-D-aspartate (NMDA)-type glutamate receptors. To further investigate the modulatory effects of zinc on NMDA receptors of different subunit compositions, we coexpressed the recombinant subunit NR1 with NR2A and/or NR2B in HEK 293 cells. In whole-cell patch-clamp recordings from these transfected cells, zinc inhibited peak glutamate-evoked current responses in a noncompetitive manner, but there were significant differences between the receptor subtypes in sensitivity to zinc inhibition. For NR1/NR2A, approximately 40% of the peak current was inhibited by zinc in a voltage-independent manner with an IC50 value of 5.0 +/- 1.6 nM and at a V(H) value of -60 mV; the remainder was blocked at a second, voltage-dependent site with an IC50 value of 79 +/- 18 microM. In contrast, NR1/NR2B currents showed nearly complete inhibition at a voltage-independent site with an IC50 value of 9.5 +/- 3.3 microM. Cells cotransfected with NR1, NR2A, and NR2B showed zinc sensitivity intermediate between that characteristic of NR1/NR2A and that of NR1/NR2B. Furthermore, zinc accelerated the macroscopic desensitization of both NR1/NR2A and NR1/NR2B in a dose-dependent manner, apparently independently of glycine-sensitive desensitization and Ca2(+)-dependent inactivation; maximal effects were to decrease desensitization time constants for NR1/NR2A by approximately 75% and for NR1/NR2B by approximately 90%. Differential modulation of NR1/NR2A and NR1/NR2B currents by zinc may play a role in regulating NMDA receptor-induced synaptic plasticity and neurotoxicity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Brain / physiology
  • Brain / ultrastructure
  • Cells, Cultured
  • Electrophysiology
  • Glutamic Acid / metabolism
  • Glutamic Acid / pharmacology
  • Mice
  • Receptors, N-Methyl-D-Aspartate / antagonists & inhibitors*
  • Receptors, N-Methyl-D-Aspartate / classification
  • Receptors, N-Methyl-D-Aspartate / physiology*
  • Recombinant Proteins / antagonists & inhibitors
  • Recombinant Proteins / classification
  • Sensitivity and Specificity
  • Transfection
  • Zinc / metabolism
  • Zinc / pharmacology*
  • Zinc / physiology

Substances

  • Receptors, N-Methyl-D-Aspartate
  • Recombinant Proteins
  • Glutamic Acid
  • Zinc