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Research ArticleNEUROPHARMACOLOGY

Iron-Mediated Free Radical Injury in Ethanol-Exposed Mouse Neural Crest Cells

Shao-yu Chen and Kathleen K. Sulik
Journal of Pharmacology and Experimental Therapeutics July 2000, 294 (1) 134-140;
Shao-yu Chen
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Kathleen K. Sulik
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Abstract

Previous studies using cell and whole embryo cultures have shown that free radicals play an important role in the ethanol-induced death of mouse neural crest cells (NCCs; a significant cell type with respect to the genesis of alcohol-related birth defects). This investigation was spurred by reports of increased iron in ethanol-exposed fetuses and the knowledge that iron can initiate the production of reactive oxygen species. Initially, the ameliorative potential of two iron chelators, deferoxamine and phenanthroline, relative to ethanol-induced cell death was examined. Cotreatment of cultured NCCs with 100 mM ethanol and either 1 or 10 μM deferoxamine or 10, 50, or 250 μM phenanthroline significantly increased the percentage of viable cells as compared with exposure to 100 mM ethanol alone. These data indicate that iron is involved in the ethanol-induced cytotoxicity. To support this premise, the direct toxicity of iron to NCCs was also examined. As expected, loading the cells with Fe(II)/Fe(III) using 8-hydroxyquinoline as a carrier had an adverse effect on their viability as did treatment with a neurotoxin, 6-hydroxydopamine, that releases iron from ferritin storage. Cotreatment with an antioxidant,N-acetylcysteine, significantly diminished the toxicity of ethanol alone, that resulting from iron loading, as well as from the combination of ethanol exposure and iron loading. These results confirm the role of free radical-mediated damage in ethanol-induced cytotoxicity and highlight the potential role of iron relative to the genesis of alcohol-related birth defects.

Footnotes

  • Send reprint requests to: Kathleen K. Sulik, Department of Cell Biology and Anatomy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7090. E-mail: mouse{at}med.unc.edu

  • ↵1 This work was supported by National Institutes of Health Grant AA11605 from the National Institute of Alcohol Abuse and Alcoholism.

  • Abbreviations:
    NCCs
    neural crest cells
    DFX
    deferoxamine
    PHE
    phenanthroline
    8HQ
    8-hydroxyquinoline
    6-OHDA
    6-hydroxydopamine
    Nac
    N-acetylcysteine
    FAS
    fetal alcohol syndrome
    DMEM
    Dulbecco's modified Eagle's medium
    • Received December 16, 1999.
    • Accepted March 27, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 294 (1)
Journal of Pharmacology and Experimental Therapeutics
Vol. 294, Issue 1
1 Jul 2000
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Research ArticleNEUROPHARMACOLOGY

Iron-Mediated Free Radical Injury in Ethanol-Exposed Mouse Neural Crest Cells

Shao-yu Chen and Kathleen K. Sulik
Journal of Pharmacology and Experimental Therapeutics July 1, 2000, 294 (1) 134-140;

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Research ArticleNEUROPHARMACOLOGY

Iron-Mediated Free Radical Injury in Ethanol-Exposed Mouse Neural Crest Cells

Shao-yu Chen and Kathleen K. Sulik
Journal of Pharmacology and Experimental Therapeutics July 1, 2000, 294 (1) 134-140;
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