250 - Proteomic study of 6-OHDA-induced neurodegeneration of rat nigrostriatal pathway

...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.