Abstract
Astrocytes and neurons cultured from mouse cerebellum and cerebral cortex were analyzed with respect to content and synthesis of amino acids as well as export of metabolites to the culture medium and the response to fluorocitrate, an, inhibitor of aconitase. The intracellular levels of amino acids were similar in the two astrocytic populations. The release of citrate, lactate and glutamine, however, was markedly higher from cerebellar than from cortical astrocytes. Neurons contained higher levels of glutamate, aspartate and GABA than astrocytic cultures. Cortical neurons were especially high in GABA and aspartate, and the level of aspartate increased specifically when the extracellular level of glutamine was elevated. Fluorocitrate inhibited the TCA cycle in the astrocytes, but was less effective in cerebellar neurons. Whereas neurons responded to fluorocitrate with an increase in the formation of lactate, reflecting, glycolysis, astrocytes decreased the formation of lactate in the presence of fluorocitrate, indicating that astrocytes to a high degree synthesize pyruvate and hence lactate from TCA cycle intermediates.
Similar content being viewed by others
References
Patel, A. J., and Hunt, A. 1985. Concentration of free amino acids in primary cultures of neurones and astrocytes. J. Neurochem. 44: 1816–1821.
Schousboe, A., Drejer, J., Hansen, G. H., and Meier, E. 1985. Cultured neurons as model systems for biochemical and pharmacological studies on receptors for neurotransmitter amino acids. Dev. Neurosci. 7:252–262.
Lehmann, A., and Hansson, E. 1987. Amino acid content in astroglial primary cultures from different brain regions during cultivation. Neurochem. Res. 12:797–800.
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. J. Neurochem. 45:335–344.
Kurz, G. M., Wiesinger, H., and Hamprecht, B. 1993. Purification of cytosolic malic enzyme from bovine brain, generation of monoclonal antibodies, and immunocytochemical localization of the enzyme in glial cells of neural primary cultures. J. Neurochem. 60:1467–1474.
Sonnewald, U., Westergaard, N., Krane, J., Unsgård, G., Petersen, S. B., and Schousboe, A. 1991. First direct demonstration of preferential release from astrocytes of citrate using [13C] NMR spectroscopy of cultured neurons and astrocytes. Neurosci. Lett. 128: 235–239.
Peters, R. A. 1957. Mechanisms of the toxicity of the active constitutent ofDichapetalum cymosum and related compounds. Adv. Enzymol. 18:113–159.
Clarke, D. D. 1991. Fluoroacetate and fluorocitrate: Mechanism of action. Neurochem. Res. 16:1055–1058.
Paulsen, R. E., Contestabile, A., Villani, L., and Fonnum, F. 1987. An in vivo model for studying function of brain tissue temporarily devoid of glial cell metabolism. The use of fluorocitrate. J. Neurochem. 48:1377–1385.
Hassel, B., Paulsen, R. E., Johnsen, A., and Fonnum, F. 1992. Selective inhibition of glial cell metabolism in vivo by fluorocitrate. Brain Res. 576:120–124.
Hertz, E., Yu, A. C. H., Hertz, L., Juurlink, B. H. J., and Schousboe, A. 1989. Preparation of primary cultures of mouse cortical neurons. Pages 183–186,in A. Shahar, J de Vellis, A. Vernadakis, and B. Haber (eds.). A dissection and tissue culture manual for the nervous system. Alan R. Liss, New York.
Hertz, L., Juurlink, B. H. J., Hertz, E., Fosmark, H., and Schousboe, A. 1989. Preparation of primary cultures of mouse (rat) astrocytes. Pages 105–108,in A. Shahar, J. de Vellis, A. Vernadakis, and B. Haber (eds.) A dissection and tissue culture manual for the nervous system. Alan R. Liss, New York.
Schousboe, A., Meier, E., Drejer, J., and Hertz, J. 1989. Preparation of primary cultures of mouse (rat) cerebellar granule cells. Pages 203–206,in A. Shahar, J. de Vellis, A. Vernadakis, and B. Haber (eds.) A dissection and tissue culture manual for the nervous system. Alan R. Liss, New York.
Drejer, J., and Schousboe, A. 1989. Selection of a pure cerebellar granule cell culture by kainate treatment. Neurochem. Res. 14: 751–754.
Larsson, O. M., Drejer, J., Kvamme, E., Svenneby, G., and Hertz, L. 1985. Ontogenetic development of glutamate and GABA metabolizing enzymes in cultured cerebral cortex interneurons and in cerebral cortex in vivo. Int. J. Dev. Neurosci. 3:177–185.
Schousboe, A., and Hertz, L. 1987. Primary cultures of GABAergic and glutamatergic neurons as model systems to study neurotransmitter functions II. Pages 33–42,in A. Vernadakis, A. Privat, J. Lauder, P. S. Timiras, and E. Giacobini (eds.) Model systems of development and ageing of the nervous system. Martinus Nijhoff Publ., Boston.
Drejer, J., Larson, O. M., Kvamme, E., Svenneby, G., Hertz, L., and Schousboe, A. 1985. Ontogenetic development of glutamate metabolizing enzymes in cultured cerebellar granule cells and in cerebellum in vivo. Neurochem. Res. 10:49–62.
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.
Martinez-Hernandez, A., Bell, K. P., and Norenberg, M. D. 1977. Glutamine synthetase: glial localization in brain. Science 195: 1356–1358.
Hornfeldt, C. S. and Larson, A. A. 1990. Seizures induced by fluoroacetic acid and fluorocitric acid may involve chelation of divalent cations in the spinal cord. Eur. J. Pharmacol. 179:307–313.
Leprince, P., Lefebvre, P. P., Rigo, J.-M., Delrée, P., Rogister, B., and Moonen, G. 1989. Cultured astroglia release a neurotoxic activity that is not related to the excitotoxins. Brain Res. 502:21–27.
Hassel, B., Sonnewald, U., Unsgård, G., and Fonnum, F. 1994. NMR spectroscopy of cultured astrocytes. Effects of glutamine and the gliotoxin fluorocitrate. J. Neurochem. 62:2187–2194.
Pasantes-Morales, H., and Schousboe, A. 1988. Volume regulation in astrocytes: A role for taurinc as an osmoeffector. J. Neurosci. Res. 20:505–509.
Slivka, A., Mytilineou, C., and Cohen, G. 1987. Histochemical evaluation of glutathione in brain. Brain Res. 409:275–284.
Ottersen, O. P., and Storm-Mathisen, J. 1985. Different neuronal localization of aspartate-like and glutamate-like immunoreactivities in the hippocampus of rat, guinea-pig and senegalese baboon (Papio papio), with a note on the distribution of γ-aminobutyrate. Neuroscience 6:589–606.
Goldberg, M. P., Monyer, H., and Choi, D. W. 1988. Hypoxic neuronal injury in vitro depends on extracellular glutamine. Neurosci. Lett. 94:52–57.
Monyer, H., and Choi, D. W. 1990. Glucose deprivation neuronal injury in vitro is modified by withdrawal of extracellular glutamine. J. Cereb. Blood Flow Metab. 10:337–342.
Sonnewald, U., Westergaard, N., Krane, J., Unsgård, G., Petersen, S. B., and Schousboe, A. 1993. Metabolism of [U-13C]glutamate in astrocytes studied by13C NMR spectroscopy: incorporation of more label into lactate than into glutamine demonstrates the importance of the tricarboxylic acid cycle. J. Neurochem. 61:1179–1182.
Kurz, G. M., Wiesinger, H., and Hamprecht, B. 1993. Immunocytochemical localization of cytosolic malic enzyme in slices from adult brain. (Abstr.) J. Neurochem. 61 (Suppl):S228D.
Meister, A. 1988. Glutathione metabolism and its selective modification. J. Biol. Chem. 263:17205–17208.
Yu A. C. H., Drejer, J., Hertz, L., and Schousboe, A. 1983. Pyruvate carboxylase activity in primary cultures of astrocytes and neurons. J. Neurochem. 41:1484–1487.
Patel, M. S. 1974. The relative significance of CO2-fixing enzymes in the metabolism of rat brain. J. Neurochem. 22:717–724.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hassel, B., Westergaard, N., Schousboe, A. et al. Metabolic differences between primary cultures of astrocytes and neurons from cerebellum and cerebral cortex. Effects of fluorocitrate. Neurochem Res 20, 413–420 (1995). https://doi.org/10.1007/BF00973096
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00973096