Prevention of ischemia/reperfusion-induced alterations in synaptosomal membrane-associated proteins and lipids by N-tert-butyl-α-phenylnitrone and difluoromethylornithine

N. C. Hall, J. M. Carney, M. Cheng, D. A. Butterfield

Research output: Contribution to journalArticlepeer-review

32 Scopus citations


Previous studies in our laboratory demonstrated the alteration in the physical state of synaptosomal membrane lipids and proteins in ischemia/ reperfusion injury using selective spin labels and electron paramagnetic resonance spectroscopy [Hall et al. (1995) Neuroscience61, 84-89]. Since many investigations have provided evidence for free radical generation during ischemia/reperfusion injury, we investigated whether a free radical scavenger would prevent the membrane damage, in gerbils. Further, experiments to determine if a secondary effect of polyamine generation at 14 h reperfusion could be blocked by this free radical scavenger or by an inhibitor of ornithine decaryboxylase were also carried out. The alterations in synaptosomal membrane integrity observed during ischemia/reperfusion injury were selectively neutralized by treatment with the free radical spin trap N-tert-butyl-α-phenylnitrone or an inhibitor of ornithine decarboxylase, difluoromethylornithine. Administration of N-tert-butyl-α-phenylnitrone prior to ischemia totally abrogated both lipid and protein alterations observed at 1 and 14 h reperfusion. Pretreatment with difluoromethylornithine neutralized only the 14 h change in lipid label motion. Treatment with N-tert-butyl-α-phenylnitrone at 6 h post ischemia showed only a slight attenuation of the 14 h lipid effect and no change in the protein effect. Difluoromethylornithine treatment at 6 h post ischemia negated the 14 h ischemia/reperfusion injury-induced lipid effect and had no effect on the protein change. These data support previous suggestions that free radicals and polyamines play a critical role in neuronal damage and cell loss following ischemia/reperfusion injury and that the polyamine effect is dependent upon free radical generation during ischemia/reperfusion injury.

Original languageEnglish
Pages (from-to)591-600
Number of pages10
Issue number2
StatePublished - Nov 1995

Bibliographical note

Funding Information:
Acknowledgements--This work was supported by a grant from NIH (AG-10836) and a research contract from Centaur Pharmaceuticals.


  • 2,2,6,6-tetramethyl-4-maleimidopiperidine-1-oxyl
  • 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
  • DFMO
  • EDTA
  • EGTA
  • EPR
  • HWHH
  • IRI
  • MAL-6
  • N-methyl-d-aspartate
  • N-tert-butyl-α-phenylnitrone
  • NMDA
  • ODC
  • PBN
  • PMSF
  • S-adenosylmethionine decarboxylase
  • W/W
  • brain
  • difluoromethylornithine
  • electron paramagnetic resonance
  • ethylenediaminetetra-acetate
  • ethyleneglycoltetraacetic acid
  • free radicals
  • gerbil
  • half width at half height of the low-field line of the 5-NS EPR spectrum
  • ischemia reperfusion injury
  • ornithine decarboxylase
  • phenylmethylsulfonyl fluoride
  • polyamines
  • the ratio of the amplitudes of the weakly immobilized to the strongly immobilized components of the low-field line of the MAL-6 EPR spectrum

ASJC Scopus subject areas

  • Neuroscience (all)


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