Mitochondrial production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate

L. L. Dugan, S. L. Sensi, L. M T Canzoniero, S. D. Handran, S. M. Rothman, T. S. Lin -, M. P. Goldberg, D. W. Choi

Research output: Contribution to journalArticle

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Abstract

Increasing evidence suggests that glutamate neurotoxicity is partly mediated by reactive oxygen species, formed as a consequence of several processes, including arachidonic acid metabolism and nitric oxide production. Here we used an oxidation-sensitive indicator, dihydrorhodamine 123, in combination with confocal microscopy, to examine the hypothesis that electron transport by neuronal mitochondria may be an important source of glutamate- induced reactive oxygen species (ROS). Exposure to NMDA, but not kainate, ionomycin, or elevated potassium stimulated oxygen radical production in cultured murine cortical neurons, demonstrated by oxidation of nonfluorescent dihydrorhodamine 123 to fluorescent rhodamine 123. Electron paramagnetic resonance spectroscopy studies using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a radical-trapping agent, also showed production of ROS by cortical neurons after NMDA but not kainate exposure. NMDA-induced ROS production depended on extracellular Ca2+, and was not affected by inhibitors of nitric oxide synthase or arachidonic acid metabolism. The increased production of ROS was was blocked by inhibitors of mitochondrial electron transport, rotenone or antimycin, and mimicked by the electron transport uncoupler, carbonyl cyanide p-trifluoromethoxyphenylhydrazone. These data support the possibility that NMDA receptor-mediated, Ca2+-dependent uncoupling of neuronal mitochondrial electron transport may contribute to the oxidative stress initiated by glutamate exposure.

Original languageEnglish (US)
Pages (from-to)6377-6388
Number of pages12
JournalJournal of Neuroscience
Volume15
Issue number10
StatePublished - 1995

Fingerprint

N-Methylaspartate
Reactive Oxygen Species
Electron Transport
Neurons
Glutamic Acid
Kainic Acid
Arachidonic Acid
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Rhodamine 123
Rotenone
Ionomycin
Electron Spin Resonance Spectroscopy
N-Methyl-D-Aspartate Receptors
Confocal Microscopy
Nitric Oxide Synthase
Spectrum Analysis
Potassium
Mitochondria
Nitric Oxide
Oxidative Stress

Keywords

  • dihydrorhodamine
  • electron transport
  • excitotoxicity
  • free radicals
  • glutamate
  • mitochondria
  • neurotoxicity
  • oxidation

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Dugan, L. L., Sensi, S. L., Canzoniero, L. M. T., Handran, S. D., Rothman, S. M., Lin -, T. S., ... Choi, D. W. (1995). Mitochondrial production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate. Journal of Neuroscience, 15(10), 6377-6388.

Mitochondrial production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate. / Dugan, L. L.; Sensi, S. L.; Canzoniero, L. M T; Handran, S. D.; Rothman, S. M.; Lin -, T. S.; Goldberg, M. P.; Choi, D. W.

In: Journal of Neuroscience, Vol. 15, No. 10, 1995, p. 6377-6388.

Research output: Contribution to journalArticle

Dugan, LL, Sensi, SL, Canzoniero, LMT, Handran, SD, Rothman, SM, Lin -, TS, Goldberg, MP & Choi, DW 1995, 'Mitochondrial production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate', Journal of Neuroscience, vol. 15, no. 10, pp. 6377-6388.
Dugan LL, Sensi SL, Canzoniero LMT, Handran SD, Rothman SM, Lin - TS et al. Mitochondrial production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate. Journal of Neuroscience. 1995;15(10):6377-6388.
Dugan, L. L. ; Sensi, S. L. ; Canzoniero, L. M T ; Handran, S. D. ; Rothman, S. M. ; Lin -, T. S. ; Goldberg, M. P. ; Choi, D. W. / Mitochondrial production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate. In: Journal of Neuroscience. 1995 ; Vol. 15, No. 10. pp. 6377-6388.
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