Abstract
L-2-Hydroxyglutaric acid (LGA) accumulates and is the biochemical hallmark of the neurometabolic disorder L-2-hydroxyglutaric aciduria (LHGA). Although this disease is predominantly characterized by severe neurological findings and pronounced cerebral atrophy, the pathomechanisms of brain injury are virtually unknown. In the present study, we investigated the effect of LGA (0.1–1 mM) on various parameters of the glutamatergic system, namely the basal and potassium-induced release of L-[3H]glutamate by synaptosomal preparations, Na+-dependent L-[3H]glutamate uptake by synaptosomal preparations and Na+-independent L-[3H]glutamate uptake by synaptic vesicles, as well as of L-[3H]glutamate binding to synaptic plasma membranes from cerebral cortex of male adult Wistar rats. We observed that LGA significantly increased L-[3H]glutamate uptake into synaptosomes and synaptic vesicles, without altering synaptosomal glutamate release and glutamate binding to synaptic plasma membranes. Although more comprehensive studies are necessary to evaluate the exact role of LGA on neurotransmission, our findings do not support a direct excitotoxic action for LGA. Therefore, other abnormalities should be searched for to explain neurodegeneration of LHGA.
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Barbot, C., Fineza, I., Diogo, L., Maia, M., Melo, J., Guimarães, A., Pires, M.M., Cardoso, M.L., and Vilarinho, L. (1997). L-2-Hydroxyglutaric aciduria: Clinical, biochemical and magnetic resonance imaging in six portuguese pediatric patients. Brain Dev. 19:268-273.
Barth, P.G., Hoffman, G.F., Jaeken, J.J., Lehnert, W., Hanefeld, F., van Gennip, A.H., Duran, M., Valk, J., Schutgens, R.B., Trefz, F.K., Reimann, G., and Hartung, H.P. (1992). L-2-Hydroxyglutaric acidaemia: A novel inherited neurometabolic disorder. Ann. Neurol. 32:66-71.
Barth, P.G., Hoffmann, G.F., Jaeken, J.J., Wanders, R.J., Duran, M., Jansen, G.A., Jakobs, C., Lehnert, W., Hanefeld, F., Valk, J., Schutgens, R.B.H., Trefz, F.K., Hartung, H.-P., Chamoles, N.A., Sfaello, Z., and Caruso, U., (1993). L-2-Hydroxyglutaric acidemia: Clinical and biochemical findings in 12 patients and preliminary report on L-2-hydroxyacid dehydrogenase. J. Inherit. Metab. Dis. 16:753-761.
Beal, M.F. (1992). Does impairment of energy metabolism, result in excitotoxic neuronal death in neurodegenerative illnesses? Ann. Neurol. 31:119-130.
Chen, E., Nyhan, W.L., Jakobs, C., Greco, C.M., Barkovich, A.J., Cox, V.A., and Packman, S. (1996). L-2-Hydroxyglutaric aciduria: Neuropathological correlations and first report of severe neurodegenerative disease and neonatal death. J. Inherit. Metab. Dis. 19:335-343.
de Mello, C.F., Kölker, S., Ahlemeyer, B., de Souza, F.R., Fighera, M.R., Mayatepek, E., Krieglstein, J., Hoffmann, G.F., and Wajner, M. (2001). Intrastriatal administration of 3-hydroxyglutaric acid induces convulsions and striatal lesions in rats. Brain Res. 916:70-75.
Divry, P., Jakobs, C., Vianey-Saban, C., Gibson, K.M., Michelakakis, H., Papadimitriou, A., Divari, R., Chabrol, B., Cournelle, M.A., and Livet, M.O. (1993). L-2-hydroxyglutaric aciduria: two further cases. J. Inherit. Metab. Dis. 16:505-507.
Dunkley, P.R., Heath, J., Harrison, S.M., Jarvie, P.E., Glenfield, P.J., and Rostas, J.A.P. (1986). A rapid Percoll gradient procedure for isolation of synaptosomes directly from an S1 fraction: Homogeneity and morphology of subcellular fractions. Brain Res. 441:59-71.
Duran, M., Kamerling, J.P., Barkker, H.D., van Gennip, H.A., and Wadman, S.K. (1980). L-2-hydroxyglutaric aciduria: An inborn error of metabolism? J. Inherit Metab. Dis. 3:11-15.
Emanuelli, T., Antunes, V., and Souza, D.O.G. (1998). Characterization of L-[3H]glutamate binding to fresh and frozen crude plasma membranes isolated from cerebral cortex of adult rats. Biochem. Mol. Biol. Int. 44:1265-1272.
Erickson-Viitanen, S., Viitanen, P., Geiger, P.J., Yang, W.C., and Bessman, S.P. (1982). Compartmentation of mitochondrial creatine phosphokinase. I. Direct demonstration of compartmentation with the use of labeled precursors. J. Biol. Chem. 257:14395-14404.
Flott-Rahmel, B., Falter, C., Schluff, P., Fingerhut, R., Christensen, E., Jakobs, C., Musshoff, U., Fautek, J.D., Ludolph, A., and Ullrich, K. (1997). Nerve cell lesions caused by 3-hydroxyglutaric acid: A possible mechanism for neurodegeneration in glutaric acidaemia I. J. Inherit. Metab. Dis. 20:387-390.
Fujitake, J., Ishikawa, Y., Fujii, H., Nishimura, K., Hayakawa, K., Inoue, F., Terada, N., Okochi, M., and Tatsuoka, Y. (1999). L-2-Hydroxyglutaric aciduria: Two Japanese adult cases in one family. J. Neurol. 246:378-382.
Fykse, E.M., and Fonnum, F.J. (1996). Amino acid neurotransmission: Dynamics of vesicular uptake. Neurochem. Res. 21:1053-1060.
Hoffmann, G.F., Jakobs, C., Holmes, B., Mitchell, L., Becker, G., Hartung, H.P., and Nyhan, W.L. (1995). Organic acids in cerebrospinal fluid and plasma of patients with L-2-hydroxyglutaric aciduria. J. Inherit. Metab. Dis. 18:189-193.
Kölker, S., Ahlemeyer, B., Krieglstein, J., and Hoffmann, G.F. (1999). 3-Hydroxyglutaric and glutaric acids are neurotoxic through NMDA receptors in vitro. J. Inherit. Metab. Dis. 22:259-262.
Kölker, S., Ahlemeyer, B., Krieglstein, J., and Hoffmann, G.F. (2000a). Cerebral organic acid disorders induce neuronal damage via excitotoxic organic acids in vitro. Amino Acids 18:31-40.
Kölker, S., Ahlemeyer, B., Krieglstein, J., Hoffmann, G.F., (2000b). Maturation-dependent neurotoxicity of 3-hydroxyglutaric and glutaric acids in vitro: A new pathophysiologic approach to glutaryl-CoA dehydrogenase deficiency. Pediatr. Res. 47:495-503.
Kölker, S., Köhr, G., Ahlemeyer, B., Okun, J.G., Pawlak, V., Horster, F., Mayatepek, E., Krieglstein, J., and Hoffmann, G.F. (2002a). Ca2+ and Na+ dependence of 3-hydroxyglutarate-induced excitotoxicity in primary neuronal cultures from chick embryo telencephalons. Pediatr. Res. 52(2):199-206.
Kölker, S., Okun, J.G., Ahlemeyer, B., Wyse, A. T., Horster, F., Wajner, M., Kohlmuller, D., Mayatepek, E., Krieglstein, J., and Hoffmann, G.F. (2002b). Chronic treatment with glutaric acid induces partial tolerance to excitotoxicity in neuronal cultures from chick embryo telencephalons. J. Neurosci. Res. 685:424-431.
Latini, A., Scussiato, K., Rosa, R.B., Leipnitz, G., Llesuy, S., Belló-Klein, A., Dutra-Filho, C.S., and Wajner, M. (in press) Induction of oxidative stress by L-2-hydroxyglutaric acid in rat brain. J. Neurosci. Res.
Lima, T.T.F, Begnini, J., Bastiani, J, Fíalo, D.B., Jurach, A., Ribeiro, M.C., Wajner, M., and de Mello, C.F. (1998). Pharmacological evidence for GABAergic and glutaminergic involvement in the convulsant and behavioral effects of glutaric acid. Brain Res. 802:55-60.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265-275.
Ludolph, A.C., Riepe, M., and Ullrich, K. (1993). Excitotoxicity, energy metabolism and neurodegeneration. J. Inherit. Metab. Dis. 16:716-723.
Meldrum, B., and Garthwaite, J. (1990). Excitatory amino acid neurotoxicity and neurodegenerative disease, Trends Pharmacol. Sci. 11:379-387.
Migues, P.V., Leal, R.B., Mantovani, M., Nicolau, M., Gabilan, N.H. (1999). Synaptosomal glutamate release induced by the fraction BC2 from the venom of the sea anemone Bunadosoma caissarum. NeuroReport 10:67-70.
Nagi, A.K., Shuster, T.A., and Delgado-Escueta, A.V. (1986). Ecto-ATPase of mammalian synaptosomes: Identification and enzymic characterization. J. Neurochem. 47:976-986.
Naito, S., and Ueda, T. (1985). Characterization of glutamate uptake into synaptic vesicles. J. Neurochem. 44:99-109.
Novelli, A., Reilly, J.A., Lysko, P.G., and Haennebery, R.C. (1998). Glutamates becomes neurotoxic via the NMDA receptor when intracellular energy levels are reduced. Brain Res. 451:205-212.
Olney, J.W. (1980). Excitotoxic mechanisms of neurotoxicity. In (P.S. Spencer, and H.H. Schaumburg, eds.), Clinical and Experimental Neurotoxicology, Williams & Wilkins, Baltimore, MD, pp. 272-294.
Ozawa, S., Kamyiar, K., and Tsuzuki, K. (1998). Glutamate receptors in the mamalian cerebral nervous system. Prog. Neurobiol. 54:581-618.
Porciúncula, L.O., Dal-Pizzol, A., Jr., Tavares, R.G., Coitinho, A.S., Emanuelli, T., Souza, D.O., and Wajner, M. (2000). Inhibition of synaptosomal [3H]glutamate and [3H]glutamate binding to plasma membranes from brain of young rats by glutaric acid in vitro. J. Neurol. Sci. 173:93-96.
Robinson, M.B., and Dowd, L.A. (1997). Heterogeneity and functional properties of subtypes of sodium-dependent glutamate transporters in the mammalian central nervous system. Adv. Pharmacol. 37:69-115.
Silva, C.G., Bueno, A.R.F., Schuck, P.F., Leipnitz, G., Ribeiro, C.A.J., Wannmacher, C.M.D., Wyse, A.T.S., and Wajner, M. (2003). L-2-Hydroxyglutaric acid inhibits mitochondrial creatine kinase activity from cerebellum of developing rats. Int. J. Dev. Neurosci. 21:217-224.
Steeghs, K., Benders, A., Oerlemans, F., de Haan, A., Heerschap, A., Ruitenbeek, W., Jost, C., van Deursen, J., Perryman, B., Pette, D., Bruckwilder, M., Koudijs, J., Jap, P., Veerkamp, J., and Wieringa, B. (1997). Altered Ca++ response in muscles with combined mitochondrial and cytosolic creatine kinase deficiencies. Cell 89:93-103.
Ullrich, K., Flott-Rahmel, B., Schluff, P., Musshoff, U., Das, A., Lücke, T., Steinfeld, R., Christensen, E., Jacobs, C., Ludolph, A., Neu, A., and Röper, R. (1999). Glutaric aciduria type I: Pathomechanisms of neurodegeneration. J. Inherit. Metab. Dis. 22:392-403.
Wolosker, H., Souza, D.O.G., and De Meis, L.J. (1996). Regulation of glutamate transport into synaptic vesicles by a chloride and proton gradient. J. Biol. Chem. 271:11726-11731.
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Junqueira, D., Brusque, A.M., Porciúncula, L.O. et al. Effects of L-2-Hydroxyglutaric Acid on Various Parameters of the Glutamatergic System in Cerebral Cortex of Rats. Metab Brain Dis 18, 233–243 (2003). https://doi.org/10.1023/A:1025559200816
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DOI: https://doi.org/10.1023/A:1025559200816