• biochemical characterization
• endoplasmic reticulum stress
• neurodegeneration
• Parkinson's disease
• post-translational modifications
• proteins transfection

Parkinson’s disease is an irreversible and progressive neurodegenerative movement disorder. In term of pathophysiology, it is considered an alpha-synucleinopathy for the abnormal accumulation of alpha-synuclein (alphaS) protein in the brain, in the form of insoluble inclusions called Lewy Bodies or Lewy Neurites. alphaS is a small protein (140 aminoacid residues), natively unfolded with a propensity to aggregate. Although it is still under debate whether transient oligomers, accumulating along the ER membrane, precede fibrils formation, several evidences demonstrate that are oligomeric intermediates to be actually cytotoxic and not the aggregates. The aim of the project was to evaluate the toxic impact of ER/M associated-alphaS oligomers in different cell models.
The aggregates population, obtained from spinal cord of alphaS transgenic mice that express human mutant A53T alphaS, was subdivided in two separate fractions with differential centrifugation according to their molecular weight and they were undergone biochemical characterization by screening with western blot. The aggregates were then administered to cells, a stable and inducible cell model expressing wild-type alphaS or to mouse primary neuronal culture directly in the medium, in order to evaluate the cytotoxic effects.
As regard the stable and inducible cell model, the aggregates were not able to enter the cells, contrarily as regard the primary neuronal culture. In fact, in this latter case, the alphaS aggregates administered were internalized by the cells and there they triggered the formation of alphaS endogenous aggregates, that ultimately led to cells death.