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bcl-2 Protects SK-N-SH Cells From 6-Hydroxydopamine Induced Apoptosis by Inhibition of Cytochrome c Redistribution

  • Richard C. Dodel
  • Kelly R. Bales
  • Mark Bender
  • Gabriel Nunez
  • Paul Hyslop
  • Wolfgang H. Oertel
  • Steven M. Paul
  • Yansheng Du

Abstract

Neurotoxicity induced by 6-hydroxydopamine (6-OHDA) is, in part, due to the production of reactive oxygen/nitrogen species (RNOS) and/or an inhibition of mitochondrial function. However, little is known about the ensuing intracellular events which ultimately result in cell death. Here we show that exposure to relatively low concentrations of 6-OHDA induce apoptosis of SK-N-SH cells. Western blots of cytosolic extracts from 6-OHDA treated cells reveal a translocation of cytochrome c from mitochondria into the cytosol. To further delineate the pathway by which 6-OHDA causes apoptosis, we investigated the effects on cell survival in bcl-2 overexpressing SK-N-SH cells and the relation to translocation of cytochrome c and free radical production. Overexpressing bcl-2 in SK-N-SH cells shifted the toxicity as well as the release of cytochrome c to higher concentrations of 6-OHDA. At concentrations of 6-OHDA in SK-N-SH-bcl-2 cells where a release of cytochrome c and subsequent cell death occurs, an increase of the oxidation of the fluorescent dye dihydrorhodamine-123 was detectable. This RNOS production paralleled the release of cytochrome c and was inhibited by bcl-2 as well. Prom these experiments, we hypothesize that cytochrome c might interact with 6-OHDA to promote an oxidative burst, which can be inhibited by bcl-2.

Keywords

Cell Death Cerebellar Granule Neuron Fluorescein Diacetate Subsequent Cell Death Initiate Lipid Peroxidation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 2000

Authors and Affiliations

  • Richard C. Dodel
    • 1
    • 4
  • Kelly R. Bales
    • 2
  • Mark Bender
    • 2
  • Gabriel Nunez
    • 3
  • Paul Hyslop
    • 2
  • Wolfgang H. Oertel
    • 4
  • Steven M. Paul
    • 1
    • 2
  • Yansheng Du
    • 1
  1. 1.Departments of Pharmacology, Toxicology, and PsychiatryIndiana University School of MedicineIndianapolisUSA
  2. 2.Neuroscience Discovery ResearchEli Lilly & Co.IndianapolisUSA
  3. 3.Department of Pathology and Comprehensive Cancer CenterThe University of Michigan Medical SchoolAnn ArborUSA
  4. 4.Department of NeurologyPhilipps-University MarburgMarburgGermany

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