Oxidative metabolism of dopamine (DA) appears to be involved in several, intersecting pathways underlying Parkinson’s disease (PD) pathogenesis (1). We have recently isolated an enzymatic activity from human midbrain tissue, by means of a specific in gel-staining procedure, set out by us, using DA and hydrogen peroxide as substrates (2). Mass spectrometry analysis of the DA peroxidising activity revealed the presence of proteins that may play a role in neurodegenerative disease, highlighting a possible functional link among DA redox cycle and protein metabolism. Pursuing the aim to untangle the intricate pathway of DA oxidation, we tested the peroxidative activity staining in rat brain tissue homogenate, in order to make use of our method of investigation in PD animal models. Distinct rat brain areas were analysed: striatum, substantia nigra, brain stem, frontal lobe, cerebellum. A red-orange activity gel band, similar to that previously developed from human midbrain tissue (2), appeared in all the rat brain regions selected. Reactivity of striatum and substantia nigra were expected, brain stem and frontal lobe showed sharp and faint colour reaction, respectively. Remarkable enzymatic activity was also developed from cerebellum. Mass spectrometry analysis of the specific activity bands is now in progress, in order to identify the proteins responsible of the enzymatic reaction. Present results appear to validate our procedure, providing an additional and useful approach to the study of oxidative metabolism of DA.
Dopamine peroxidation: a novel study from rat brain areas
DE IULIIS, ANGELA;ARRIGONI, GIORGIO;VIANELLO, FABIO;GIUSTI, PIETRO;ARSLAN, PAOLA
2009
Abstract
Oxidative metabolism of dopamine (DA) appears to be involved in several, intersecting pathways underlying Parkinson’s disease (PD) pathogenesis (1). We have recently isolated an enzymatic activity from human midbrain tissue, by means of a specific in gel-staining procedure, set out by us, using DA and hydrogen peroxide as substrates (2). Mass spectrometry analysis of the DA peroxidising activity revealed the presence of proteins that may play a role in neurodegenerative disease, highlighting a possible functional link among DA redox cycle and protein metabolism. Pursuing the aim to untangle the intricate pathway of DA oxidation, we tested the peroxidative activity staining in rat brain tissue homogenate, in order to make use of our method of investigation in PD animal models. Distinct rat brain areas were analysed: striatum, substantia nigra, brain stem, frontal lobe, cerebellum. A red-orange activity gel band, similar to that previously developed from human midbrain tissue (2), appeared in all the rat brain regions selected. Reactivity of striatum and substantia nigra were expected, brain stem and frontal lobe showed sharp and faint colour reaction, respectively. Remarkable enzymatic activity was also developed from cerebellum. Mass spectrometry analysis of the specific activity bands is now in progress, in order to identify the proteins responsible of the enzymatic reaction. Present results appear to validate our procedure, providing an additional and useful approach to the study of oxidative metabolism of DA.
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