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Volume 17, Number 15, Issue of August 1, 1997 pp. 5711-5725
Copyright ©1997 Society for Neuroscience

High-Affinity Zinc Inhibition of NMDA NR1-NR2A Receptors

Received March 31, 1997; revised May 8, 1997; accepted May 13, 1997.

Pierre Paoletti, Philippe Ascher, and Jacques Neyton

Laboratoire de Neurobiologie, Centre National de la Recherche Scientifique Unité de Recherche Associée 1857, Ecole Normale Supérieure, 75005 Paris, France

Micromolar concentrations of extracellular Zn²⁺ are known to antagonize native NMDA receptors via a dual mechanism involving both a voltage-independent and a voltage-dependent inhibition. We have tried to evaluate the relative importance of these two effects and their subunit specificity on recombinant NMDA receptors expressed in HEK 293 cells and Xenopus oocytes. The comparison of NR1a-NR2A and NR1a-NR2B receptors shows that the voltage-dependent inhibition is similar in both types of receptors but that the voltage-independent inhibition occurs at much lower Zn²⁺ concentrations in NR1a-NR2A receptors (IC₅₀ in the nanomolar range) than in NR1a-NR2B receptors (IC₅₀ in the micromolar range). The high affinity of the effect observed with NR1a-NR2A receptors was found to be attributable mostly to the slow dissociation of Zn²⁺ from its binding site. By analyzing the effects of Zn²⁺ on varied combinations of NR1 (NR1a or NR1b) and NR2 (NR2A, NR2B, NR2C), we show that both the NR1 and the NR2 subunits contribute to the voltage-independent Zn²⁺ inhibition. We have observed further that under control conditions, i.e., in zero nominal Zn²⁺ solutions, the addition of low concentrations of heavy metal chelators markedly potentiates the responses of NR1a-NR2A receptors, but not of NR1a-NR2B receptors. This result suggests that traces of a heavy metal (probably Zn²⁺) contaminate standard solutions and tonically inhibit NR1a-NR2A receptors. Chelation of a contaminant metal also could account for the rapid NR2A subunit-specific potentiations produced by reducing compounds like DTT or glutathione.

Key words: NMDA; zinc; DTT; heavy metals; recombinant receptors; ionic channels

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				Z.-g. Xiong, K. A. Pelkey, W. Y. Lu, Y. M. Lu, J. C. Roder, J. F. MacDonald, and M. W. Salter Src Potentiation of NMDA Receptors in Hippocampal and Spinal Neurons Is Not Mediated by Reducing Zinc Inhibition J. Neurosci., November 1, 1999; 19(21): RC37 - RC37. [Abstract] [Full Text] [PDF]

				I. Joshi and L.-Y. Wang Developmental profiles of glutamate receptors and synaptic transmission at a single synapse in the mouse auditory brainstem J. Physiol., March 15, 2002; (2002) 2001013506. [Abstract] [PDF]

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