Inducible over-expression of wild type alpha-synuclein in human neuronal cells leads to caspase-dependent non-apoptotic death

J Neurochem. 2009 Jun;109(5):1348-62. doi: 10.1111/j.1471-4159.2009.06054.x. Epub 2009 Mar 23.

Abstract

Alpha-synuclein (ASYN) is central in Parkinson's disease pathogenesis. Converging pieces of evidence suggest that the levels of ASYN expression play a critical role in both familial and sporadic Parkinson's disease. To elucidate the mechanism underlying wild type (WT) ASYN-mediated neurotoxicity, we have generated a novel Tet-Off SHSY-5Y cell line, conditionally expressing WT ASYN. Induction of human WT ASYN in retinoic acid-differentiated SHSY-5Y cells leads to accumulation of soluble ASYN oligomers, in the absence of inclusions, and to gradual cellular degeneration. Morphologically, the death observed is non-apoptotic. Caspases other than caspase 3, including caspase 9, are activated and caspase inhibition diminishes death by acting at a point upstream of cytochrome c release. Application of Scyllo-inositol, an oligomer-stabilizing compound, prevents neuronal death in this model. These findings are consistent with a model in which oligomeric ASYN triggers the initial activation of the apoptotic pathway, which is however blocked downstream of the mitochondrial checkpoint, thus leading to a death combining in a unique fashion both apoptotic and non-apoptotic features. This novel inducible cell model system may prove valuable in the deciphering of WT ASYN-induced pathogenic effects and in the assessment and screening of potential therapeutic strategies.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology
  • Apoptosis / drug effects*
  • Apoptosis / physiology
  • Caspases / metabolism*
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cytochromes c / metabolism
  • Enzyme Inhibitors / pharmacology
  • Gene Expression Regulation / drug effects
  • Green Fluorescent Proteins / genetics
  • Humans
  • Inositol / pharmacology
  • Membrane Potential, Mitochondrial / drug effects
  • Mitochondria / drug effects
  • Neuroblastoma / ultrastructure
  • Staurosporine / pharmacology
  • Time Factors
  • Transfection / methods
  • Tretinoin / pharmacology
  • alpha-Synuclein / genetics
  • alpha-Synuclein / metabolism*
  • bcl-X Protein / metabolism

Substances

  • Amino Acid Chloromethyl Ketones
  • Enzyme Inhibitors
  • alpha-Synuclein
  • bcl-X Protein
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • scyllitol
  • Inositol
  • Tretinoin
  • Cytochromes c
  • Caspases
  • Staurosporine