title: The effects of copper ions on glutamate receptors in cultured rat cortical neurons.
abstract: Copper plays an important role in the function of many physiological processes and can affect different neurotransmitter systems. In this study, we used the patch-clamp technique to investigate the effect of copper ions on glutamate receptors in cultured rat cortical neurons. Cu2+ inhibited (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptors with an IC50 of 4.3 +/- 0.6 microM (with 100 microM kainate, holding potential -60 mV). The concentration-response could be best described by a two-site binding model. Moreover, copper reduced the efficacy of kainate at the AMPA receptor: in the presence of 30 microM Cu2+, the EC50 of kainate was shifted from 100.3 +/- 2.0 microM to 329.9 +/- 31.4 microM. The block by copper ions was not use-dependent. Complete recovery only occurred after the application of a high agonist concentration, or in the presence of the antioxidant dithiotreitol (DTT). A high concentration of histidine, a physiological ligand for Cu2+, did not augment the recovery. The kinetics of block were compared to those induced by 2,3-dihydro-6-nitro-7-sulfamoyl-benz(F)quinoxaline (NBQX), a well-described competitive antagonist of AMPA receptors. The onset, as well as the offset of block by NBQX could be well approximated by single exponential functions with time constants of 0.28 +/- 0.02 and 0.87 +/- 0.09 s, respectively. Within seconds of wash-out of the antagonist, the response to kainate completely recovered. The kinetics of copper block were more complex: the block developed more slowly, and the onset, as well as the offset could be described by two exponential functions with quite different time constants (tau(on1), 0.8 +/- 0.13 s; tau(on2), 8.32 +/- 1.13 s; tau(off1), 0.17 +/- 0.01 s; tau(off2), 69 +/- 36.3 s). In addition to the described effects, Cu2+ also blocked currents induced by the application of N-methyl-D-aspartate (IC50: 15.0 +/- 2.6 microM with 50 microM NMDA). Based on these findings, a modulatory role of copper ions on the neurotransmission by excitatory amino acids is discussed.
authors.affiliation: Department of Biological Research, Boehringer Ingelheim KG, Germany. 1015742177@COMPUSERVE.COM
authors.forename: T
authors.initials: T
authors.lastname: Weiser
authors.forename: M
authors.initials: M
authors.lastname: Wienrich
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects*; D003300:Copper / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D007608:Kainic Acid / Q000494:pharmacology; D051381:Rats; D017470:Receptors, Glutamate / Q000187:drug effects*
chemical_list: D017470:Receptors, Glutamate; D003300:Copper; D007608:Kainic Acid