Abstract
Cell cultures are useful tools to study the mechanisms involved in cell death following hypoxia or ischemia. By manipulating the extracellular environment, conditions that closely mimic the conditions that are thought to occur in vivo can be produced. These conditions permit study of cell’s reaction to the trauma under specific conditions. Monitoring of the extracellular pH and ionic environment in cell cultures is much easier than in vivo. Further, metabolites produced by injured cells can be quantitated easier from cultures than from tissues in vivo.
Cell cultures have recently been used to examine in detail the neurotoxicity of glutamate. Intracellular Ca2+ increases appear to be involved in the mechanisms of neurotoxic cell death. This Ca2+ entry appears to be through the NMDA receptor’s Ca2+ channel. Ischemic and hypoxic injury produced by mechanisms other than glutamate neurotoxicity appear to involve increases in intracellular Ca2+ by releasing internal Ca2+ stores or by the influx of extracellular Ca2+. This Ca2+ entry may be through voltage-gated channels of the NMDA channel, or may be attributable to membrane perturbations. Through the use of cell cultures, each of the mechanism’s involvement in the injury can be delineated.
Similar content being viewed by others
References
Benveniste H., Drejer J., Schoisboe A., and Diemer N. H. (1984) Elevation of the extracellular concentrations of glutamate and aspartate in rat hippocampus during transient cerebral ischemia monitored by intracellular microdialysis.J. Neurochem. 43, 1369–1374.
Callahan D. J., Engle M. J., and Volpe J. J. (1990) Hypoxic injury to developing glial cells: Protective effect of high glucose.Ped. Res. 27, 186–190.
Chan P. H., Chu L., and Chen S. (1990) Effects of MK-801 on glutamate-induced swelling of astrocytes in primary cell culture.J. Neurosci. Res. 25, 87–93.
Choi D. W. (1985) Glutamate neurotoxicity in cortical cell culture is calcium dependent.Neurosci. Lett. 58, 293–297.
Choi D. W., Maulucci-Gedde M., and Kriegstein A. R. (1987) Glutamate neurotoxicity in cortical cell culture.J. Neurosci. 7, 357–368.
Demediuk P., Saunders R. D., Anderson D. K., Means E. D., and Horrocks L. A. (1985) Membrane lipid changes in laminectomized and traumatized cat spinal cord.Proc. Natl. Acad. Sci. USA 82, 7071–7075.
Demediuk P., Daly M. P., and Faden A. I. (1989) Effect of impact trauma on neurotransmitter and nonneurotransmitter amino acids in rat spinal cord.J. Neurochem. 52, 1529–1536.
Drejer J., Benveniste H., Diemer N. H., and Schousboe A. (1985) Cellular origin of ischemia-induced glutamate release from brain tissue in vivo and in vitro.J. Neurochem. 45, 145–151.
Frandsen A. and Schousboe A. (1991) Dantrolene prevents glutamate cytotoxicity and Ca2+ release from intracellular stores in cultured cerebral cortical neurons.J. Neurochem. 56, 1075–1078.
Freese A., DiFiglia M., Koroshetz W. J., Beal M. F., and Martin J. B. (1990) Characterization and mechanism of glutamate neurotoxicity in primary striatal cultures.Brain Res. 521, 254–264.
Giffard R. G., Monyer H., and Choi D. W. (1990) Selective vulnerability of cultured cortical glia to injury by extracellular acidosis.Brain Res. 530, 138–141.
Goldberg W. J., Kadingo R. M., and Barrett J. N. (1986) Effects of ischemia-like conditions on cultured neurons: Protection by low Na+, low Ca2+ solutions.J. Neurosci. 6, 3144–3151.
Goldberg M. P., Pham P. C., and Choi D. W. (1987a) Dextrorphan and dextromethorphan attenuate hypoxic injury in neuronal culture.Neurosci. Lett. 80, 11–15.
Goldberg M. P., Weiss J. H., Pham P. C., and Choi D. W. (1987b) N-methyl-D-aspartate receptors mediate hypoxic neuronal injury in cortical culture.J. Pharmacol. Exp. Therapeut. 243, 784–791.
Goldberg M. P., Viseskul V., and Choi D. W. (1988) Phencyclidine receptor ligands attenuate cortical neuronal injury after N1-methyl-d-aspartate exposure or hypoxia.J. Pharmacol. Exp. Therapeut. 245, 1081–1087.
Gregory G. A., Yu A. C. H., and Chan P. H. (1989) Fructose-1,6-bisphospate protects astrocytes from hypoxic damage.J. Cereb. Blood Flow Metab. 9, 29–34.
Gregory G. A., Welsh F. A., Yu A. C. H., and Chan P. H. (1990) Fructose-1,6-bisphosphate reduces ATP loss from hypoxic astrocytes.Brain Res. 516, 310–312.
Horrocks L. A., Dorman R. V., and Porcellati G. (1984) Fatty acids and phosolipids in brain during ischemia, inCerebral Ischemia, International Congress Series No. 654 (Bes A., Braquet P., Paoletti R., and Siesjö B. K., eds), pp. 221–222, Elsevier, Amsterdam.
Ikeda M., Yoshida S., Busto R., Santiso M., and Ginsberg M. D. (1986) Polyphosphoinositides as a probable source of brain free fatty acids accumulated at the onset on ischemia.J. Neurochem. 47, 123–132.
Kimelberg H. K., Goderie S. K., Higman S., Pang S., and Waniewski R. A. (1990) Swelling-induced release of glutamate, aspartate, and taurine from astrocyte cultures.J. Neurosci. 10, 1583–1591.
Michaels R. L. and Rothman S. M. (1990) Glutamate neurotoxicity in vitro: Antogonist pharmacology and intracellular calcium concentrations.J. Neurosci. 10, 283–292.
Monyer H., Goldberg M. P., and Choi D. W. (1989) Glucose deprivation neuronal injury in cortical culture.Brain Res. 483, 347–354.
Morales H. P. and Schousboe A. (1988) Volume regulation in astrocytes: A role for taurine as an osmoeffector.J. Neurosci. Res. 20, 505–509.
Murphy E. J., Anderson D. K., Means E. D., and Horrocks L. A. (1989) Pressure induced trauma in ROC-1 oligodendroglial cell cultures.Trans. Am. Soc. Neurochem. 20, 141.
Norenberg M. D., Mozes L. W., Gregorios J. B., and Norenberg L. O. B. (1987) Effects of lactic acid on astrocytes in primary culture.J. Neuropathol. Exp. Neurol. 46, 154–166.
Panter S. S., Yum S. W., and Faden A. I. (1990) Alteration in extracellular amino acids after traumatic spinal cord injury.Ann. Neurol. 27, 96–99.
Phillips H. J. (1973) Dye exclusion tests for cell viability, inTissue Culture Methods and Applications (Kruse P. P., Jr. and Patterson M. K., eds.), pp. 406–408, Academic, New York.
Renkawek K. Herbaczynska-Cedro K., and Mossakowski M. J. (1986) The effect of prostacyclin on the morphological and enzymatic properties of CNS cultures exposed to anoxia.Acta Neurol. Scand. 73, 111–118.
Rotman B. and Papermaster B. W. (1966) Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters.Proc. Natl. Acad. Sci. USA 55, 134–141.
Singh N. P. and Stephens R. E. (1986) A novel technique for viable cell determinations.Stain Tech. 61, 315–318.
Swanson R. A., Yu A. C. H., Chan P. H., and Sharp F. R. (1990) Glutamate increases glycogen content and reduces glucose utilization in primary astrocyte culture.J. Neurochem. 54, 490–495.
Weiss J. H., Hartley D. M., Koh J., and Choi D. W. (1990) The calcium channel blocker nifedipine attenuates slow excitatory amino acid neurotoxicity.Science 247, 1474–1477.
Yoshida S., Inoh S., Asano T., Sano K., Kubota M., Shmazaki H., and Ueta N. (1980) Effect of transient ischemia on free fatty acids and phospholipids in the gerbil brain.J. Neurosurg. 53, 323–331.
Yu A. C. H., Gregory G. A., and Chan P. H. (1989) Hypoxia-induced dysfunction and injury of astrocytes in primary cell cultures.J. Cereb. Blood Flow Metab. 9, 20–28.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Murphy, E.J., Horrocks, L.A. Mechanisms of hypoxic and ischemic injury. Molecular and Chemical Neuropathology 19, 95–106 (1993). https://doi.org/10.1007/BF03160171
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF03160171