Neurochemical Research

, Volume 20, Issue 7, pp 779–784 | Cite as

The iron component of sodium nitroprusside blocks NMDA-induced glutamate accumulation and intracellular Ca2+ elevation

  • Seikwan Oh
  • Patrick P. McCaslin
Original Articles


These studies were designed to compare the effects of nitric oxide (NO) generating compounds with those of several iron containing, compounds which do not generate NO on glutamate receptor function. Stimulation of primary cultures of cerebellar granule cells with N-methyl-D-aspartate (NMDA) or kainate results in the elevation of intracellular calcium ([Ca2+]i) and cGMP and the release of glutamate. The iron containing compounds, sodium nitroprusside (SNP), potassium ferrocyanide (K4Fe(CN)6) and potassium ferricyanide (K3Fe(CN)6) decrease the NMDA-induced release of glutamate. SNP is the only compound of the above 3 agents which generates NO. A non-iron, NO generating compound, S-nitroso-N-acetylpenicillamin (SNAP), has no effect on the NMDA-induced glutamate release. Potassium ferrocyanide (Fe II), but not potassium ferricyanide (Fe III), blocks NMDA-induced cGMP elevations after 3 min exposure times. This contrasts with the NO generating compounds (both SNP and SNAP) which elevate cGMP levels. Furthermore, both potassium ferrocyanide (Fe II) and SNP (Fe II) suppress the elevation of [Ca2+]i induced by NMDA but neither potassium ferricyanide (Fe III) nor SNAP are effective in this regard. These effects are also independent of cyanide as another Fe II compound, ferrous sulfate (FeSO4) is also able to suppress NMDA-induced elevations of [Ca2+]i SNP was unable to suppress kainate receptor functions. Collectively, these results indicate that Fe II, independently of NO, has effects on NMDA receptor function.

Key Words

Cyclic GMP kainate potassium ferrocyanide potassium ferricyanide cerebellar granule cells 


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  1. 1.
    Bliss, T. V., and Collingridge, G. L. 1993. A synaptic model of memory: long-term potentiation in the hippocampus. Nature 361:31–39.PubMedGoogle Scholar
  2. 2.
    Bourne, H. R., and Nicoll, R. 1993. Molecular machines integrate coincident synaptic signals, Cell/Neuron 72/10 (supple):65–75.Google Scholar
  3. 3.
    Monaghan, D. T., and Cotman, C. W. 1985. Distribution of NMDA-sensitive L-3H-glutamate binding sites in rat brain as determined by quantitative autography. J. Neurosci. 5:2909–2919.PubMedGoogle Scholar
  4. 4.
    Johnson, J. W., and Ascher, P. 1987. Glycine potentiates the NMDA response in cultured mouse brain neurons, Nature 325:529–575.PubMedGoogle Scholar
  5. 5.
    Anis, N. A., Berry, S. C., Burton, N. R., and Lodge, D. 1983. The dissociative anaesthetics ketamins and phencyclidine, selectively reduce excitation of central mammalian neurons by N-methyl-aspartate. Br. J. Pharmacol. 79, 565.PubMedGoogle Scholar
  6. 6.
    Huettner, J. E., and Bean, B. P. 1988. Block of N-methyl-D-aspartate-activated current by the anticonvulsant MK-801: Selective binding to open channels, Proc. Natl. Acad. Sci. 85:1307–1311.PubMedGoogle Scholar
  7. 7.
    Nowak, L., Bregestovski, P., Ascher, P., Herbet, A., and Prochiantz, A. 1984. Magnesium gates glutamate-activated channels in mouse central neurons, Nature 307:462–465.PubMedGoogle Scholar
  8. 8.
    Mayer, M. L., Westbrook, G. L., and Guthrie, P. B. 1984. Voltagedependent block by Mg++ of NMDA responses in spinal cord neurons, Nature 309:261–263.PubMedGoogle Scholar
  9. 9.
    Peters, S., Koh, J., and Choi, D. W. 1987. Zinc selectively blocks the action of N-methyl-D-aspartate on cortical neurons, Science 236:589–592.PubMedGoogle Scholar
  10. 10.
    Westbrook, G. L., and Mayer, M. L. 1987. Micromolecular concentrations of Zn++ antagonize NMDA and GABA responses of hippocampal neurons, Nature 328:640–643.PubMedGoogle Scholar
  11. 11.
    Aizenman, E., Lipton, S. A., and Loring, R. H. 1989. Selective modulation of NMDA responses by reduction and oxidation, Neuron 2:1257–1263.PubMedGoogle Scholar
  12. 12.
    Kohr, G., Eckardt, S., Luddens, H., Monyer, H., and Seeberg, P. H. 1994. NMDA receptor channels: subunit-specific potentiation by reducing agents, Neuron 2:1031–1040.Google Scholar
  13. 13.
    Manzoni, O., Prezeau, L., Marin, P., Deshager, S., Bockaert, J., and Fagni, L. 1992. Nitric oxide-induced blockade of NMDA receptors, Neuron 8:653–662.PubMedGoogle Scholar
  14. 14.
    Ujihara, H., Akaike, A., Tamura, Y., Yokota, T., Sasa, M., Kashii, S., and Honda, Y. 1993. Blockade of retinal NMDA receptors by sodium nitroprusside is probably due to nitric oxide formation, Japan J. Pharmacol. 61:375–377.Google Scholar
  15. 15.
    Bredt, D. S., and Snyder, S. H. 1989. Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum, Proc. Natl Acad. Sci. 86:9030–9033.PubMedGoogle Scholar
  16. 16.
    Garthwaite, J., Charles, S. L., and Chess-Williams, R. 1988. Endothelium-derived relaxing factor release on activation of NMDA receptors suggests role as intercellular messenger in the brain, Nature 336:385–388.PubMedGoogle Scholar
  17. 17.
    McCaslin, P. P., and Morgan, W. W. 1987. Cultured cerebellar cells as anin vitro model of excitatory amino acid receptor function, Brain Res. 417:380–384.PubMedGoogle Scholar
  18. 18.
    Ellison, D. W., Beal, M. F., and Martin, J. B. 1987. Amino acid neurotransmitters in postmortem human brain analyzed by high performance liquid chromatography with electrochemical detection, J. Neurosci. Metho., 19:305–315.Google Scholar
  19. 19.
    Bradford, M. M. 1976. A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt. Biochem., 72:248–254.PubMedGoogle Scholar
  20. 20.
    Cai, Z., and McCaslin, P. P. 1992. Amitriptyline, desipramine, cyproheptadine and carbamazepine, in concentrations used therapeutically, reduce kainate- and N-methyl-D-aspartate-induced intracellular Ca2+ levels in neuronal culture, Eur. J. Pharmacol. 219:53–57.PubMedGoogle Scholar
  21. 21.
    Grynkiewicz, G., Poenie, M. F., and Tsien, R. Y. 1985. A new generation of Ca2+ indicators with greatly improved fluorescence properties, J. Biol. Chem. 260:3440–3450.PubMedGoogle Scholar
  22. 22.
    Lonart, G., Wang J. W., and Johnson, K. M. 1992. Nitric oxide induces neurotransmitter release from hippocampal slices, Eur. J. Pharmacol. 220:271–272.PubMedGoogle Scholar
  23. 23.
    Lawrence, A. J., and Jarrot, B. 1993. Nitric oxide increases interstitial excitatory amino acid release in the rat dorsomedial medulla oblongata, Neurosci, Lett. 151:126–129.Google Scholar
  24. 24.
    Hoyt, K. R., Tang, L. H., Aizenman, E., and Reynolds, I. 1992. Nitric oxide modulates NMDA-induced increases in intracellular Ca2+ in cultured rat forebrain neurons, Brain Res. 592:310–316.PubMedGoogle Scholar
  25. 25.
    Lei, S. Z., Pan, Z. H., Aggarwal, S. K., Chen, H. V., Hartman, J., Sucher, N. J., and Lipton, S. A. 1992. Effect of nitric oxide production on the redox modulatory site of the NMDA receptor-channel complex, Neuron 8:1087–1099.PubMedGoogle Scholar
  26. 26.
    Wroblewski, J. T., Kledrowski, L., Raulli, R., and Costa, E. 1992. Role of nitric oxide in signal transduction of glutamate receptors. in. Excitatory amino acids, ed. R. P. Simon, (Thieme Medical Publishers, New York) p. 81–87.Google Scholar
  27. 27.
    Southam, E., and Garthwaite, J. 1991. Comparative effects of some nitric oxide donors on cyclic GMP levels in rat cerebellar slices, Neurosci. Lett. 130:107–109.PubMedGoogle Scholar
  28. 28.
    Lipton, S. A. 1993. Prospects for clinically tolerated NMDA antagonists: Open-channel blockers and alternative redox states of nitric oxide, TINS 16:527–532.PubMedGoogle Scholar
  29. 29.
    Lipton, S. A., Choi, Y. B., Pan, Z. H., Lei, S. Z., Chen, H. V., Sucher, N. J., Loscalzo, J., Singel, D. J., and Stamler, J. S. 1993. A redox-based mechanism for the neuroprotective and neurodestructive effects of nitric oxide and related nitroso-compounds, Nature 364:626–632.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1995

Authors and Affiliations

  • Seikwan Oh
    • 1
  • Patrick P. McCaslin
    • 2
  1. 1.Department of Pharmacology and ToxicologyUniversity of Mississippi Medical CenterJackson
  2. 2.Division of ResearchOchsner Medical FoundationNew Orleans

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