Abstract
Neuronal cortical cell cultures obtained from fetal mice were subjected to an hypoxic insult produced by sodium cyanide (1 mM) for 24 h. Neurochemical assays were performed 13–14 days after plating on intact cells in situ to determine if there was a specific pattern of cellular dysfunction in addition to morphologic change. Ro5-4864-displaceable benzodiazepine (BDZ) binding and high-affinity [3H]β-alanine uptake were not reduced when compared to control values. However, specific and clonazepam-displaceable BDZ binding (81±4% and 50±9% of control values, respectively), high-affinity [3H]GABA uptake (75±2%), and choline acetyltransferase activity (82±2%) were significantly lower. When the data were expressed in terms of protein content, high-affinity [3H]β-alanine uptake was significantlyincreased in cyanide-exposed and magnesium-treated cultures (123±5% and 117±3%, respectively) as was R05-4864-displaceable BDZ binding (152±14%), consistent with stimulation of nonneuronal BDZ binding and increased glial neurotransmitter uptake. Moreover, pretreatment of the cultures with magnesium effectively prevented both the morphologic and neurochemical evidence of hypoxic injury. These data lend further support to the notion that the release of excitatory neurotransmitters may mediate neurotoxicity in developing brain.
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References
Myers, R. E. 1979. Lactic acid accumulation as a cause of brain edema and cerebral necrosis resulting from oxygen deprivation. Pages 85–114,in, Korobkin, R., and Guilleminault, D. (eds.) Advances in Perinatal Neurology, New York: Spectrum.
Raichle, M. 1983. The pathophysiology of brain ischemia. Ann. Neurol. 13:2–10.
Rehncrona, S., Rosen, I., and Siesjo, B. 1980. Excessive cellular acidosis: an important mechanism of neuronal damage in the brain. Act Physiol. Scand. 110:435–437.
Siesjo, B. 1981. Cell damage in the brain. A speculative synthesis. J. Cereb. Blood Flow Metab. 1:155–185.
Choi, D. W. 1985. Glutamate neurotoxicity in cortical cell cultures is calcium dependent. Neurosci. Lett. 58:293–297.
Johnston, M. V., and Silverstein, F. S. 1986. New insights into mechanisms of neuronal damage in developing brain. Pediat. Neurosci. 12:87–89.
Rothman, S. M. 1983. Synaptic activity mediates death of hypoxic neurons. Science 220:536–537.
Rothman, S. 1984. Synaptic release of excitatory amino acid neurotransmitters mediates anoxic neuronal death. J. Neurosci. 4:1884–1891.
Rothman, S. M., and Olney, J. W. 1986. Glutamate and the pathophysiology of hypoxic-ischemic brain damage. Ann. Neurol. 19:105–111.
Silverstein, F. S., Buchanan, K., and Johnston, M. V. 1986. Perinatal hypoxia-ischemia disrupts striatal high-affinity [3H]glutamate uptake into synaptosomes. J. Neurochem. 47:1614–1619.
Weiss, J., Goldberg, M. P., and Choi, D. W. 1986. Ketamine protects cultured neocortical neurons from hypoxic injury. Brain Res. 380:186–190.
Swaiman, K. F., Neale, E. A., Fitzgerald, S. C., and Nelson, P. G. 1982. A method for large-scale production of mouse brain cortical cultures. Dev. Brain Res. 3:361–369.
sher, P. K., Schrier, B. K., and Van Putten, D. 1982. An in situ assay for determination of benzodiazepine binding. Dev. Neurosci. 5:271–277.
Schon, F., and Kelly, J. S. 1975. Selective uptake of [3H]β-alanine by glia: association with the glial uptake system for GABA. Brain Res. 86:243–257.
Schrier, B. K., Neale, E. A., Sher, P. K., Brenneman, D. E., and Nelson, P. G. 1981. GABA uptake and other cell markers distinguish cell types in drug toxicity studies with dispersed cultures of developing CNS cells. Pages 268–279,in Okada, Y., and Roberts, E. (eds.), Problems in GABA Research, Amsterdam: Excerpta Medica, Elsevier North-Holland.
Godfrey, E. W., Schrier, B. K., and Nelson, P. G. 1980. Source and target cell specificities of a conditioned medium factor that increases choline acetyltransferase activity in cultured spinal cord cells. Dev. Biol. 77:403–418.
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.
Jones, M. G., Bicklar, D., Wilson, M. T., Brunori, M., Colosimo, A., and Sarti, M. P. 1984. A re-examination of the reactions of cyanide with cytochrome c oxidase. 1984. Biochem. J. 220:57–66.
Gallager, D. W., Mallorga, P., Oertel, W., Henneberry, R., and Tallman, J. 1981. [3H]diazepam binding in mammalian central nervous system: a pharmacological consideration. J. Neurosci. 1:218–225.
Sher, P. K. 1982. Development of the benzodiazepine receptor in cultures of fetal mouse cerebral cortex mimics its development in vivo. Dev. Neurosci. 5:263–270.
Sher, P. K., Neale, E. A., and Machen, V. L., 1986. Autoradiographic localization of the benzodiazepine receptor in cultures of fetal mouse cerebral cortex. J. Neurochem. 46:899–904.
McCarthy, K. D., and Harden, T. K. 1981. Identification of two benzodiazepine binding sites on cells cultures from rat cerebral cortex. J. Pharmacol exp. Ther. 216:183–191.
Talwar, D., and Sher, P. K. Development of the benzodiazepine receptor in murine glial cells. Dev. Neurosci. In press.
Stewart, R. M., and Rosenberg, R. N. 1979. Physiology of glia: glial-neuronal interactions. Int. Rev. Neurobiol. 21:275–309.
Rice, J. E., III, Vanucci, R. C., and Brierly, J. B. 1981. The influence of immaturity on hypoxic-ischemic brain damage in the rat. Ann. Neurol. 9:131–141.
Hertz, L. 1981. Features of astrocyte function apparently involved in the response of the central nervous system to ischemia-hypoxia. J. Cer. Blood Flow Metab. 1:143–154.
Benveniste, H., Drejer, J., Schousboe, A. and Diemer, N. H. 1984. Elevation of extracellular concentrations of glutamate and aspartate during transient cerebral ischemia. J. Neurochem. 43:1369–1374.
Hauptman, M., Nelson, D., Wilson, D. F., and Erecinska, M. 1984. Neurotransmitter aminoacids in the CNS. II. Some changes in amino acid levels in rat brain synaptosomes during and after in vitro anoxia and simulated ischemia. Brain Res. 304:23–35.
Weinstein, L. A., and Landin, D. M. D. 1985. Anoxia does not alter membrane structure in cultured astrocytes. Abst. Soc. Neurosci. 11:777.
Coyle, J. T., Bird, S. J., Evans, R. H., Gulley, R. L., Nadler, J. V., Nicklas, W. J., and Olney, J. W. 1981. Excitatory amino acid neurotoxins: selectivity, specificity, and mechanism of action. Neurosci. Res. Prog. Bull. 19:329–427.
Collins, R. L., and Olney, J. W. 1982. Focal cortical seizures cause distant thalamic lesions. Science 218:177–179.
Wieloch, T. 1985. Hypoglycemia-induced neuronal damage prevented by an N-methyl-d-aspartate antagonist. Science 230:681–683.
Meldrum, B. 1985. Possible therapeutic applications of antagonists of excitatory amino acid neurotransmitters. Clin. Sci. 68:113–122.
Aitken, P. G., and Schiff, S. J. 1986. Barbiturate protection against hypoxic neuronal damage in vitro. J. Neurosurg. 65:230–232.
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Sher, P.K. Neurochemical correlates of cyanide-induced hypoxic neuronal damage in vitro. Neurochem Res 13, 159–163 (1988). https://doi.org/10.1007/BF00973328
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DOI: https://doi.org/10.1007/BF00973328