Rotenone induces apoptosis via activation of bad in human dopaminergic SH-SY5Y cells

J Pharmacol Exp Ther. 2004 Dec;311(3):948-53. doi: 10.1124/jpet.104.071381. Epub 2004 Jul 27.

Abstract

Chronic complex I inhibition caused by rotenone induces features of Parkinson's disease in rats, including selective nigrostriatal dopaminergic degeneration and Lewy bodies with alpha-synuclein-positive inclusions. To determine the mechanisms underlying rotenone-induced neuronal death, we used an in vitro model of human dopaminergic SH-SY5Y cells. In rotenone-induced cell death, rotenone induced Bad dephosphorylation without changing the amount of Bad proteins. Rotenone also increased the amount of alpha-synuclein in cells showing morphological changes in response to rotenone. Because Bad and alpha-synuclein are known to bind to 14-3-3 proteins, we examined the effects of rotenone on these complexes. Whereas a decreased Bad amount bound to 14-3-3 proteins, rotenone increased alpha-synuclein binding to these proteins. Because dephosphorylation by calcineurin activates Bad, we examined the possible involvement of Bad activation in rotenone-induced apoptosis by using the calcineurin inhibitor tacrolimus (FK506). Tacrolimus suppressed two rotenone-induced actions: Bad dephosphorylation and apoptosis. Furthermore, the inhibition of caspase-9, which functions downstream from Bad, completely suppressed rotenone-induced apoptosis. Our findings demonstrate that Bad activation plays a role in rotenone-induced apoptosis of SH-SY5Y cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Biotransformation / drug effects
  • Blotting, Western
  • Carrier Proteins / metabolism*
  • Caspase 9
  • Caspases / metabolism
  • Cell Line
  • Cell Nucleus / drug effects
  • Cell Nucleus / ultrastructure
  • DNA Fragmentation
  • Dopamine / physiology*
  • Dose-Response Relationship, Drug
  • Electrophoresis, Agar Gel
  • Humans
  • Immunoprecipitation
  • Immunosuppressive Agents / pharmacology
  • Microscopy, Fluorescence
  • Nerve Tissue Proteins
  • Phosphorylation / drug effects
  • Rotenone / pharmacology*
  • Synucleins
  • Tacrolimus / pharmacology
  • Uncoupling Agents / pharmacology*
  • alpha-Synuclein
  • bcl-Associated Death Protein

Substances

  • BAD protein, human
  • Carrier Proteins
  • Immunosuppressive Agents
  • Nerve Tissue Proteins
  • SNCA protein, human
  • Synucleins
  • Uncoupling Agents
  • alpha-Synuclein
  • bcl-Associated Death Protein
  • Rotenone
  • CASP9 protein, human
  • Caspase 9
  • Caspases
  • Dopamine
  • Tacrolimus