Exposure of cultured cerebellar neurons to ±-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) in the presence of aniracetam protects all of the vulnerable neurons against the excitotoxic actions of glutamate acting on N-methyl-D-aspartate receptors. The protective effect of AMPA was both time- and concentration-dependent. Aniracetam alone did not protect the neurons against the excitotoxic effects of glutamate. Pretreatment of cerebellar neurons with the AMPA antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione blocked the neuroprotective effect mediated by AMPA indicating that the neuroprotective effect is mediated specifically by AMPA receptors. An excitotoxic concentration of glutamate, which killed between 60–80% of granule cell neurons on day 8in vitro, mediated its toxic effect via a time-dependent apoptotic pathway. Pretreatment of cerebellar granule cell neurons with AMPA (500 μM) completely blocked glutamate-mediated apoptosis. Our results suggest that AMPA receptors may play an important role in neuronal survival.
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Ankarcrona, M., Dypbukt, J.M., Bonfoco, E., Zhivotovsky, B., Orrenius, S., Lipton, S.A. and Nicotera, P. (1995) Glutamateinduced neuronal death: a succession of necrosis or apo-ptosis depending on mitochondrial function.Neuron 15: 961–73.
Ben-Ari, Y., Khazipov, R., Leinekugel, X., Caillard, O. and Gaiarsa, J.L. (1997) GABAA, NMDA and AMPA receptors: a developmentally regulated “menage a trois.”Trends Neurosci.20: 523–9.
Boxall, A.R. and Garthwaite, J. (1995) Synaptic excitation mediated by AMPA receptors in rat cerebellar slices is selectively enhanced by aniracetam and cyclothiazide.Neuwchem. Res. 20: 605–9.
Cebers, G. and Liljequist, S. (1995) Modulation of AMPA/ kainate receptors by cyclothiazide increases cytoplasmic free Ca2+ and45Ca2+ uptake in brain neurons.Eur. J. Pharmacol. 290: 105–15.
Choi, D.W. (1988) Calcium-mediated neurotoxicity: relationship to specific channel types and role in ischemic damage.Trends Neurosci. 11: 465–9.
Copani, A., Genazzani, A.A., Aleppo, G., Casabona, G., Canonico, P.L., Scapagnini, U. and Nicoletti, F. (1992) Nootropic drugs positively modulate a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-sensitive glutamate receptors in neuronal cultures.J. Neuwchem. 58: 1199–204.
Dessi, R, Charriaut-Marlangue, C, Khrestchatisky, M. and Ben-Ari, Y. (1993) Glutamate-induced neuronal death is not a programmed cell death in cerebellar culture.J. Neuwchem. 60: 1953–5.
Didier, M., Bursztajn, S., Adamec, E., Passani, L., Nixon, R.A., Coyle, J.T., Wei, J.Y. and Berman, S.A. (1996) DNA strand breaks induced by sustained glutamate excitotoxicity in primary neuronal cultures.J. Neurosci. 16: 2238–50.
Dipasquale, B., Marini, A.M. and Youle, R.J. (1991) Apoptosis and DNA degradation induced by 1-methyl-4-phenyl-pyridinium in neurons.Biochem. Biophys. Res. Commun. 181: 1442–8.
Favaron, M., Manev, H., Alho, H., Bertolino, M., Ferret, B., Guidotti, A. and Costa, E. (1988) Gangliosides prevent glutamate and kainate neurotoxicity in primary neuronal cultures of neonatal rat cerebellum and cortex.Proc. Natl. Acad. Sci. USA 85: 7351–5.
Gallo, V., Suiergiu, R., Giovannini, C. and Levi, G. (1987) Glutamate receptor subtypes in cultured cerebellar neurons: modulation of glutamate and gamma-aminobutyric acid release.J. Neuwchem. 49: 1801–9.
Giardina, S., Cheung, N.S., Reid, M.T. and Beart, P.M. (1998) Kainate-induced apoptosis in cultured murine cerebellar granule cells elevates expression of the cell cycle gene cyclin Dl.J. Neurochem. 71: 1325–8.
Hack, N. and Balazs, R. (1995) Properties of AMPA receptors expressed in rat cerebellar granule cells in culture.J. Neurochem. 65: 1077–84.
Hayashi Sekiyama, N., Nakanishi, S., Jane, D.E., Sunter, D.C., Birse, E.F., Udvarhelyi, P.M. and Watkins, J.C. (1994) Analysis of agonist and antagonist activities of phenylglycine derivatives for different cloned metabotropic glutamate receptor subtypes.Neurosci. 14: 3370–7.
Hoffman, P.L., Rabe, C.S., Moses, F. and Tabakoff, B. (1989) N-Methyl-D-aspartate receptors and ethanol: inhibition of calcium flux and cyclic GMP production.J. Neurochem. 52: 1937–40.
Hollmann, M., O’Shea-Greenfield, A., Rogers, S.W. and Heinemann, S. (1989) Cloning by functional expression of a member of the glutamate receptor family.Nature 342: 643–8.
Jensen, J.B., Schousboe, A. and Pickering, D.S. (1998) Development of calcium-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in cultured neocortical neurons visualized by cobalt staining.J. Neurosci. Res. 54: 273–81.
Johansen, T.H., Chaudhary, A. and Verdoorn, T.A. (1995) Interactions among GYKI-52466, cyclothiazide, and aniracetam at recombinant AMPA and kainate receptors.Mol. Pharmacol. 48: 946–55.
Kato, K., Puttfarcken, P.S., Lyons, W.E. and Coyle, J.T. (1991) Developmental time course and ionic dependence of kainate-mediated toxicity in rat cerebellar granule cell cultures.J. Pharmacol. Exp. Ther. 256: 402–11.
Keinanen, K., Wisden, W., Sommer, B., Werner, P., Herb, An., Verdoorn, T.A., Sakmann, B. and Seeburg, P.H. (1990) A family of AMPA-selective glutamate receptors.Science 249: 556–60.
Lee, J.-M., Zipfel, G.J. and Choi, D.W. (1999) The changing landscape of ischaemic brain injury mechanisms.Nature 399(Suppl.): A7-A14.
Longone, P., Impagnatiello, R, Mienville, J.M., Costa, E. and Guidotti, A. (1998) Changes in AMPA receptor-spliced variant expression and shift in AMPA receptor spontaneous desensitization pharmacology during cerebellar granule cell maturationin vitro.J. Mol. Neurosci. 11: 23–41.
Liao, D., Hessler, N.A. and Malinow, R. (1995) Activation of postsynaptically silent synapses during pairing-induced LTP in CA1 region of hippocampal slice.Nature 375: 400–04.
Marini, A.M. and Novelli, A. (1991) The glutamate uptake blocker DL-threo-3-hydroxyaspartate reduces NMDA receptor activation by glutamate in cultured neurons.European J. Pharmacol. 194: 131–2.
Marini, A.M. and Paul, S.M. (1992) N-methyl-D-aspartate receptor-mediated neuroprotection in cerebellar granule cells requires new RNA and protein synthesis.Proc. Natl. Acad. Sci. 89: 6555–9.
Marini, A.M., Rabin, S.J., Lipsky, R.H. and Mocchetti, I. (1998) Activity-dependent release of brain-derived neurotrophic factor underlies the neuroprotective effect of N-methyl-D-aspartate.J. Biol. Chem. 273: 29394–9.
Marini, A.M., Ueda, Y and June, C.H. (1999) Intracellular survival pathways: The role of intracellular calcium.New York Academy of Sciences.890: 421–437.
Muller, D. and Lynch, G. (1988) Long-term potentiation differentially affects two components of synaptic responses in hippocampus.Proc. Natl. Acad. Sci. USA 85: 9346–50.
Murphy, S.N. and Miller, J.R. (1989) Regulation of Ca++ influx into striatal neurons by kainic acid.J. Pharmacol. Exp. Ther. 249: 184–93.
Nicoletti, R, Bruno, V, Catania, M.V., Battaglia, G., Copani, A., Barbagallo, G., Cena, V., Sanches-Prieto, J., Spano, P.F. and Pizzi, M. (1999) Group-I metabotropic glutamate recepors: hypotheses to explain their dual role in neurotoxicity and neuroprotection.Neuropharmacology 38: 1477–84.
Novelli, A., Reilly J.A., Lysko, P.G. and Henneberry, R.C. (1988) Glutamate becomes neurotoxic via the N-methyl-D-aspartate receptor when intracellular energy levels are reduced.Brain Res. 451: 205–12.
Olney, J.W., Ho, O.L. and Rhee, V. (1971) Cytotoxic effects of acidic and sulphur-containing amino acids on the infant mouse central nervous system.Exp. Brain Res. 14: 61–76.
Partin, K.M., Fleck, M.W. and Mayer, M.L. (1996) AMPA receptor flip/flop mutants affecting deactivation, desensitization, and modulation by cyclothiazide, aniracetam, and thiocyanate.J. Neurosci. 16: 6634–47.
Perez-Pinzon, M.A., Maier, CM., Yoon, E.J., Sun, G.H., Giffard, R.G. and Steinberg, G.K. (1995) Correlation of CGS 19755 neuroprotection againstin vitro excitotoxicity and focal cerebral ischemia.J. Cereb. Blood Flow Metab. 15: 865–76.
Pizzi, M., Fallacara, C, Arrighi, V., Memo, M. and Spano, P.F. (1993) Attenuation of excitatory amino acid toxicity by metabotropic glutamate receptor agonists and aniracetam in primary cultures of cerebellar granule cells.J. Neurochem. 61: 683–9.
Pruss, R.M., Akeson, R.C., Racke, M.M. and Wilburn, J.L. (1991) Agonist activated cobalt identifies divalent cation-permeable kainate receptors on neurons and glial cells.Neuron 7: 509–18.
Schoepp, D.D., Jane, D.E. and Monn, J.A. (1999) Pharmacological agents acting at subtypes of metabotropic glutamate receptors.Neuropharmacology 38: 1431–76.
Sheardown, M.J., Nielsen, E.O., Hansen, A.J., Jacobsen, P. and Honore, T. (1990) 2,3-Dihydroxy-6-nitro-7-sulfamoyl-benzo-(F)quinoxaline: a neuroprotectant for cerebral ischemia.Science 247: 571–4.
Shi, S.-H., Hayashi, Y., Petralia, R.S., Zaman, S.H., Wenthold, R.J., Svoboda, K. and Malinow, R. (1999) Rapid spine delivery and redistribution of AMPA receptors after synaptic NMDA receptor activation.Science 284: 1811–16.
Swanson, G.T., Kamboj, S.K. and Cull-Candy, S.G. (1997) Single-channel properties of recombinant AMPA receptors depend on RNA editing, splice variation, and subunit composition.J. Neurosci. 17: 58–69.
Yamada, K.A. (1998) Modulating exciatory synaptic neurotransmission: potential treatment for neurological disease?Neurobiol. Dis. 5: 67–80.
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Banaudha, K., Marini, A.M. AMPA prevents glutamate-induced neurotoxicity and apoptosis in cultured cerebellar granule cell neurons. neurotox res 2, 51–61 (2000). https://doi.org/10.1007/BF03033327
- Cerebellar neurons