Kainic Acid, NMDA and Bicuculline Induce Elevation in Concentrations of Glutathione and Amino Acids in Vivo: Biomarkers for Seizure Predisposition?


The present study was carried out to investigate the effect of NMDA, bicuculline and kainic acid (KA) on the extracellular concentration of glutathione, phosphoethanolamine (PEA) and taurine in rat hippocampus in vivo. Rats were implanted with intrahippocampal microelectrodes perfused with free-glucose Krebs-Ringer solution and allowed to recover for about 2 h. After assaying baseline concentrations of amino acids, NMDA or bicuculline was administered intrahippocampally, whereas KA was given systemically. Either treatment resulted in significant high extracellular concentrations of glutathione, but only NMDA or KA resulted in high concentrations of PEA and taurine. Interestingly, the increase in glutathione concentration due to KA was followed by a delayed increase of glutamate and PEA. Our results demonstrated that increased efflux of glutathione, a common consequence of different neuroexcitotoxic agents, occurs in vivo. Given that the agents used in the present study were also convulsunts, the implication of the findings on seizure predisposition was also considered.

Share and Cite:

Abbas, A. (2015) Kainic Acid, NMDA and Bicuculline Induce Elevation in Concentrations of Glutathione and Amino Acids in Vivo: Biomarkers for Seizure Predisposition?. Journal of Behavioral and Brain Science, 5, 163-172. doi: 10.4236/jbbs.2015.55017.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Forman, H.J., Zhang, H. and Rinna, A. (2009) Glutathione: Overview of Its Protective Roles, Measurement, and Biosynthesis. Molecular Aspects of Medicine, 30, 1-12.
[2] Slivka, A., Spina, M.B. and Cohen, G. (1987) Reduced and Oxidized Glutathione in Human and Monkey Brain. Neuroscience Letters, 74, 112-118.
[3] Chinta, S.J., Kumar, M.J., Hsu, M., Rajagopalan, S., Kaur, D., et al. (2007) Inducible Alterations of Glutathione Levels in Adult Dopaminergic Midbrain Neurons Result in Nigrostriatal Degeneration. Journal of Neuroscience, 27, 13997-14006.
[4] Oliveira, A.A., Almeida, J.P.C., Freitas, R.M., Nascimento, V.S., Aguiar, L.M.V., et al. (2007) Effects of Levetiracetam in Peroxidation Level, Nitrite-Nitrate Formation and Antioxidant Enzymatic Activity in Mice Brain after Pilocarpine-Induced Seizures. Cellular and Molecular Neurobiology, 27, 395-406.
[5] Kohr, G., Eckardt, S., Lüddens, H., Monyer, H. and Seeburg, P.H. (1994) NMDA Receptor Channels: Subunit-Specific Potentiation by Reducing Agents. Neuron, 12, 1031-1040.
[6] Bodhinathan, K., Kumar, A. and Foster, T.C. (2010) Intracellular Redox State Alters NMDA Receptor Response during Aging through Ca2+/Calmodulin-Dependent Protein Kinase II. The Journal of Neuro-science, 30, 1914-1924.
[7] Ogita, K., Kitago, T., Nakamuta, H., Fukuda, Y., Koida, M., et al. (1986) Glutathione-Induced Inhibition of Na+- Independent and -Dependent Bindings of L-[3H] Glutamate in Rat Brain. Life Sciences, 39, 2411-2418.
[8] Patten, A.R., Brocardo, P.S., Sakiyama, C., Wortman, R.C., Noonan, A., et al. (2013) Impairments in Hippocampal Synaptic Plasticity Following Prenatal Ethanol Exposure Are Dependent on Glutathione Levels. Hippocampus, 23, 1463-1475.
[9] Robillard, J.M., Gordon, G.R., Choi, H.B., Christie, B.R. and MacVicar, B.A. (2011) Glutathione Restores the Mechanism of Synaptic Plasticity of the Adult. PLoS ONE, 6, e20676.
[10] Bruce, A.J. and Baudry, M. (1995) Oxygen Free Radicals in Rat Limbic Structures after Kainite-Induced Seizures. Free Radical Biology and Medicine, 18, 993-1002.
[11] Freitas, R.M., Vasconcelos, S.M.M., Souza, F.C.F., Viana, G.S.B. and Fonteles, M.M.F. (2005) Oxidative Stress in the Hippocampus after Pilocarpine-Induced Status Epilepticus in Wistar Rats. FEBS Journal, 272, 1307-1312.
[12] Dugan, L.L., Sensl, S.L., Canzoniero, L.M.T., Handran, S.D., Rothman, S.M., et al. (1995) Mitochondrial Production of Reactive Oxygen Species in Cortical Neurons Following Exposure to N-methyl-D-Aspartate. Journal of Neuroscience, 15, 6377-6388.
[13] Wade, J.V., Olson, J.P., Samson, F.E., Nelson, S.R. and Pazdernik, T.L. (1988) A Possible Role for Taurine in Osmoregulation within the Brain. Journal of Neurochemistry, 51, 740-745.
[14] El-Abhar, H.S. and El Gawad, H.M. (2003) Modulation of Cortical Nitric Oxide Synthase, Glutamate, and Redox State by Nifedipine and Taurine in PTZ-Kindled Mice. Epilepsia, 44, 276-281.
[15] Huxtable, R.J. (1992) Physiological Actions of Taurine. Physiological Reviews, 72, 101-163.
[16] Hagberg, H., Lehmann, A., Sandberg, M., Nystrom, B., Jacobson, I., et al. (1985) Ischemia-Induced Shift of Inhibitory and Excitatory Amino Acids from Intra- to Extracellular Compartments. Journal of Cerebral Blood Flow & Metabolism, 5, 413-419.
[17] Sandberg, M., Butcher, S.P. and Hagberg, H. (1986) Extracellular Overflow of Neuroactive Amino Acids during Severe Insulin-Induced Hypoglycemia: In Vivo Dialysis of the Rat Hippocampus. Journal of Neurochemistry, 47, 178-184.
[18] Lo, E.H., Bosque-Hamilton, P. and Meng, W. (1998) Inhibition of Poly(ADP-Ribose) Polymerase: Reduction of Ischemic Injury and Attenuation of N-methyl-D-Aspartate-Induced Neurotransmitter Dysregulation. Stroke, 29, 830-836.
[19] Wallin, C., Weber, S.G. and Sandberg, M. (1999) Glutathione Efflux Induced by NMDA and Kainate: Implications in Neurotoxicity? Journal of Neurochemistry, 73, 1566-1572.
[20] Wallin, C., Abbas, A.K., Tranberg, M., Weber, S.G., Wigstrom, H. and Sandberg, M. (2003) Searching for Mechanisms of N-Methyl-D-Aspartate-Induced Glutathione Efflux in Organotypic Hippocampal Cultures. Neurochemical Research, 28, 281-291.
[21] Lehmann, A., Isacsson, H. and Hamberger, A. (1983) Effects of in Vivo Administration of Kainic Acid on the Extracellular Amino Acid Pool in the Rabbit Hippocampus. Journal of Neurochemistry, 40, 1314-1320.
[22] Keillor, J.W., Castonguay, R. and Lherbet, C. (2005) Gamma-Glutamyl Transpeptidase Substrate Specificity and Catalytic Mechanism. Methods in Enzymology, 401, 449-467.
[23] Li, X., Wallin, C., Weber, S.G. and Sandberg, M. (1999) Net Efflux of Cysteine, Glutathione and Related Metabolites from Rat Hippocampal Slices during Oxygen/Glucose Deprivation: Dependence on Gamma-Glutamyl Transpeptidase. Brain Research, 815, 81-88.
[24] Kim, H.C., Jhoo, W.K., Kim, W.K., Suh, J.H., Shin, E.J., et al. (2000) An Immunocytochemical Study of Mitochondrial Manganese-Superoxide Dismutase in the Rat Hippocampus after Kainite Administration. Neuroscience Letters, 281, 65-68.
[25] Shih, Y.H., Chein, Y.C., Wang, J.Y. and Fu, Y.S. (2004) Ursolic Acid Protects Hippocampal Neurons against Kainite-Induced Excitotoxicity in Rats. Neuroscience Letters, 362, 136-140.
[26] Saito, T., Sakamoto, K., Koizumi, K. and Stewart, M. (2006) Repeatable Focal Seizure Suppression: A Rat Preparation to Study Consequences of Seizure Activity Based on Urethane Anesthesia and Reversible Carotid Artery Occlusion. Journal of Neuroscience Methods, 155, 241-250.
[27] Fedele, E., Varnier, G. and Paiteri, M. (1997) In Vivo Microdialysis Study of GABAA and GABAB Receptors Modulating the Glutamate Receptor/NO/Cyclic GMP Pathway in the Rat Hippocampus. Neuropharmacology, 36, 1405-1415.
[28] Anschel, D.J., Ortega, E. and Fisher, R.S. (2004) Diazepam Prophylaxis for Bicuculline-Induced Seizures: A Rat Dose-Response Model. Neuroscience Letters, 356, 66-68.
[29] Engel Jr., J. (1992) Experimental Animal Models of Epilepsy: Classification and Relevance to Human Epileptic Phenomena. Epilepsy Research. Supplement, 8, 9-20.
[30] Tranberg, M., Stridh, M.H., Guy, Y., Jilderos, B., Wigstrom, H., Weber, S.G. and Sandberg, M. (2004) NMDA-Receptor Mediated Efflux of N-Acetylaspartate: Physiological and/or Pathological Importance? Neurochemistry International, 45, 1195-1204.
[31] de Freitas, R.M. (2010) Lipoic Acid Alters δ-Aminolevulinic Dehydratase, Glutathione Peroxidase and Na+,K+-ATPase Activities and Glutathione-Reduced Levels in Rat Hippocampus after Pilocarpine-Induced Seizures. Cellular and Molecular Neurobiology, 30, 381-387.
[32] Li, Z.X., Yu, H.M. and Jiang, K.W. (2013) Tonic GABA Inhibition in Hippocampal Dentate Granule Cells: Its Regulation and Function in Temporal Lobe Epilepsies. Acta Physiologica, 209, 199-211.
[33] Eyo, U.B., Peng, J., Swiatkowski, P., Mukherjee, A., Bispo, A. and Wu, L.J. (2014) Neuronal Hyperactivity Recruits Microglial Processes via Neuronal NMDA Receptors and Microglial P2Y12 Receptors after Status Epilepticus. Journal of Neuroscience, 34, 10528-10540.
[34] Zeevalk, G.D., Bernard, L.P., Sinha, C., Ehrhart, J. and Nicklas, W.J. (1998) Excitotoxicity and Oxidative Stress during Inhibition of Energy Metabolism. Developmental Neuroscience, 20, 444-453.
[35] Wu, J.Y., Wu, H., Jin, Y., Wie, J., Sha, D., et al. (2009) Mechanism of Neuroprotective Function of Taurine. Advances in Experimental Medicine and Biology, 643, 169-179.
[36] Lehmann, A., Hagberg, H., Jacobson, I. and Hamberger, A. (1985) Effects of Status Epilepticus on Extracellular Amino Acids in the Hippocampus. Brain Research, 359, 147-151.
[37] Saransaari, P. and Oja, S.S. (2000) Taurine Release Modified by GABAergic Agents in Hippocampal Slices from Adult and Developing Mice. Amino Acids, 18, 17-30.
[38] Gasull, T., Sarri, E., DeGregorio-Rocasolano, N. and Trullas, R. (2003) NMDA Receptor Overactivation Inhibits Phospholipid Synthesis by Decreasing Choline-Ethanolamine Phosphotransferase Activity. Journal of Neuroscience, 23, 4100-4107.
[39] Stein, B.A. and Sapolsky, R.M. (1988) Chemical Adrenalectomy Reduces Hippocampal Damage Induced by Kainic Acid. Brain Research, 473, 175-180.
[40] Cavus, I., Kasoff, W.S., Cassaday, M.P., Jacob, R., Gueorguieva, R., et al. (2005) Extracellular Metabolites in the Cortex and Hippocampus of Epileptic Patients. Annals of Neurology, 57, 226-235.
[41] Lévesque, M., Avoli, M. and Bernard, C. (2015) Animal Models of Temporal Lobe Epilepsy Following Systemic Chemoconvulsant Administration. Journal of Neuroscience Methods (In Press).
[42] Ludvig, N. and Tang, H.M. (2000) Cellular Electrophysiological Changes in the Hippocampus of Freely Behaving Rats during Local Microdialysis with Epileptogenic Concentration of N-Methyl-D-Aspartate. Brain Research Bulletin, 51, 233-240.
[43] Stein, A.G., Eder, H.G., Blum, D.E., Drachev, A. and Fisher, R.S. (2000) An Automated Drug Delivery System for Focal Epilepsy. Epilepsy Research, 39, 103-114.
[44] Jiang, D., Akopian, G., Ho, Y.S., Walsh, J.P. and Andersen, J.K. (2000) Chronic Brain Oxidation in a Glutathione Peroxidase Knockout Mouse Model Results in Increased Resistance to Induced Epileptic Seizures. Experimental Neurology, 164, 257-268.
[45] Fico, A., Manganelli, G., Cigliano, L., Bergamo, P., Abrescia, P., et al. (2008) 2-deoxy-d-Ribose Induces Apoptosis by Inhibiting the Synthesis and Increasing the Efflux of Glutathione. Free Radical Biology and Medicine, 45, 211-217.
[46] Regan, R.F. and Guo, Y.P. (1999) Potentiation of Excitotoxic Injury by High Concentrations of Extracellular Reduced Glutathione. Neuroscience, 91, 463-470.
[47] Aroniadou-Anderjaska, V., Pidoplichko, V.I., Figueiredo, T.H., Almeida-Suhett, C.P., Prager, E.M. and Braga, M.F.M. (2012) Presynaptic Facilitation of Glutamate Release in the Basolateral Amygdala: A Mechanism for the Anxiogenic and Seizurogenic Function of GluK1 Receptors. Neuroscience, 221, 157-169.
[48] Sierra-Paredes, G., Loureiro, A.I., Wright, L.C., Sierra-Marcuo, G. and Soares-da-Silva, P. (2014) Effects of Eslicarbazepine Acetate on Acute and Chronic Latrunculin A-Induced Seizures and Extracellular Amino Acid Levels in the Mouse Hippocampus. BMC Neuroscience, 15, 134.
[49] Oja, S.S. and Saransaari, P. (2013) Regulation of Taurine Release in the Hippocampus of Developing and Adult Mice. Advances in Experimental Medicine and Biology, 775, 135-143.
[50] Junyent, E., Utrera, J., Romero, R., Pallas, M., Camins, A., Duque, D. and Auladell, C. (2009) Prevention of Epilepsy by Taurine Treatments in Mice Experimental Model. Journal of Neuroscience Research, 87, 1500-1508.
[51] Durelli, L. and Mutani, R. (1983) The Current Status of Taurine in Epilepsy. Clinical Neuropharmacology, 6, 37-48.
[52] Dzirkale, Z., Pupure, J., Rumaks, J., Svirskis, S., Vanina, M., et al. (2011) Comparative Study of Taurine and Tauropyrone: GABA Receptor Binding, Mitochondrial Processes and Behavior. Journal of Pharmacy and Pharmacology, 63, 230-237.
[53] Seu, K.J., Cambrea, L.R., Everly, R.M. and Hovis, J.S. (2006) Influence of Lipid Chemistry on Membrane Fluidity: Tail and Headgroup Interactions. Biophysical Journal, 91, 3727-3735.
[54] Lehmann, A. and Hamberger, A. (1984) A Possible Interrelationship between Extracellular Taurine and Phosphoethanolamine in the Hippocampus. Journal of Neurochemistry, 42, 1286-1290.
[55] Huxtable, R.J., Crosswell, S. and Parker, D. (1989) Phospholipid Composition and Taurine Content of Synaptosomes in Developing Rat Brain. Neurochemistry International, 15, 233-238.
[56] Jirsa, V.K., Stacey, W.C., Quilichini, P.P., Ivanov, A.I. and Bernard, C. (2014) On the Nature of Seizure Dynamics. Brain, 137, 2210-2230.
[57] Klein, J.A. and Ackerman, S.L. (2003) Oxidative Stress, Cell Cycle, and Neurodegeneration. Journal of Clinical Investigation, 111, 785-793.
[58] Currais, A. and Maher, P. (2013) Functional Consequences of Age-Dependent Changes in Glutathione Status in the Brain. Antioxidants & Redox Signaling, 19, 813-822.
[59] Bragin, D.E., Zhou, B., Ramamoorthy, P., Müller, W.S., Connor, J.A. and Shi, H. (2010) Differential Changes of Glutathione Levels in Astrocytes and Neurons in Ischemic Brains by Two-Photon Imaging. Journal of Cerebral Blood Flow & Metabolism, 30, 734-738.
[60] Meyerson, B.A., Linderoth, B., Karlsson, H. and Ungerstedt, U. (1990) Microdialysis in the Human Brain: Extracellular Measurements in the Thalamus of Parkinsonian Patients. Life Sciences, 46, 301-308.
[61] Hillered, L., Persson, L., Pontén, U. and Ungerstedt, U. (1990) Neurometabolic Monitoring of the Ischaemic Human Brain Using Microdialysis. Acta Neurochirurgica, 102, 91-97.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.