Differential sensitivity of recombinant N-methyl-D-aspartate receptor subtypes to zinc inhibition.
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.
Chen
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Department of Psychiatry, University of British Columbia, Vancouver, Canada.
Moshaver
A
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Raymond
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LA
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000502:physiology / Q000648:ultrastructure; D002478:Cells, Cultured; D004594:Electrophysiology; D018698:Glutamic Acid / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors* / Q000145:classification / Q000502:physiology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000145:classification; D012680:Sensitivity and Specificity; D014162:Transfection; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology* / Q000502:physiology
D016194:Receptors, N-Methyl-D-Aspartate; D011994:Recombinant Proteins; D018698:Glutamic Acid; D015032:Zinc