250 - Studio proteomico della neurodegenerazione indotta da 6-OHDA nel circuito nigrostriatale di ratto
Autore/i: A. De Iuliis, J. Grigoletto, A. Recchia, P. Giusti, P. Arslan
Rivista: RIMeL - IJLaM, Vol. 2, N. 3, 2006 (MAF Servizi srl ed.)
...In an attempt to identify some of the proteins that are
involved in dopaminergic neuronal death, we used the
proteomic methods to analyze this animal model of
PD.
Methods: Five hemiparkinsonian rats were obtained by
intranigral stereotaxic injection of 6-OHDA.The right
6-OHDA-lesioned substantia nigra and striatum tissues
along with the left, unlesioned controlateral tissues, were
excised and homogenized, using urea-based buffer, to
extract the tissues protein.
The separation of the protein mixtures and the visualization
of the protein patterns obtained were performed
using two-dimensional polyacrylamide gel
electrophoresis (2D-PAGE).
Protein profiles of control and treated tissues were
compare by the PDQuest 2D-gel analysis software
(BIO-Rad Lab., Hercules, CA, USA). The protein spots
showing differential expression were analysed by matrix
assisted laser desorption/ionizing time of flight
(MALDI-TOF) mass spectrometry.
Results: The brain protein extraction and solubilization
protocol was validated obtaining a satisfactory protein
profile. In comparison to the normal rats, emiparkinsonian
animals exhibited a different expression in alpha-
enolase and beta-actin in substantia nigra and striatum,
respectively.
Conclusion: The proteomic study of rats unilateral injection
of 6-OHDA into the nigro-striatal tissues allowed
us to identify two proteins, alpha-enolase and betaactin,
showing increased levels in the 6-OHDA-lesioned
brain tissues compared to control. Previous studies
described the same proteins as oxidized and proteins
in Alzheimer´s disease (AD) brain. Our preliminary
data could mirror those results pointing out a
common mechanism of neurodegenerative diseases.