BURTÉ, F., 2009. Proteomic analysis of cellular models of neurodegeneration and mitochondrial dysfunction. PhD, Nottingham Trent University.
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Mitochondrial dysfunction is thought to contribute to neurodegenerative processes. As an example, dysfunction of complex I of the electron transport chain has been observed in Parkinson’s disease patients and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP), a complex I inhibitor, produces a Parkinsonian state in mammals. The aims of the present study were to determine the effects of MPTP on the mitochondrial proteome in a cellular model using mouse N2a neuroblastoma cells and to identify novel biomarkers of MPTP-induced toxicity. The enrichment of mitochondria and the presence of cross-contamination from other subcellular components were monitored using a range of molecular markers. Mitochondrial proteins were then fractionated using an optimised 2-dimensional gel electrophoresis (2DE) protocol and the reproducibility of the method was investigated. A preliminary study comparing the mitochondrial proteome profile from two different states of mouse N2a neuroblastoma cells, mitotic and differentiated, was undertaken to establish whether differentiation of cells had major effects on the mitochondrial proteome. Since nine proteins showed changes in levels, which included stress-70 protein and aconitase, it was decided that differentiation did affect the mitochondrial proteome: hence, differentiated cells were used for further studies. The effects of different concentrations (0 to 5 mM) and time-points (0 to 48 hours) of MPTP on plasma membrane integrity, cellular metabolic activity, cellular ATP concentration, mitochondrial potential, cytochrome c release and a variety of caspase activities were investigated.
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|Divisions:||Schools > School of Science and Technology|
|Depositing User:||EPrints Services|
|Date Added:||09 Oct 2015 09:33|
|Last Modified:||09 Oct 2015 09:33|
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