Oxidative metabolism of dopamine (DA) appears to beinvolved in several, intersecting pathways underlyingParkinson’s disease (PD) pathogenesis (1). We have recentlyisolated an enzymatic activity from human midbrain tissue,by means of a specific in gel-staining procedure, set out byus, using DA and hydrogen peroxide as substrates (2). Massspectrometry analysis of the DA peroxidising activityrevealed the presence of proteins that may play a role inneurodegenerative disease, highlighting a possiblefunctional link among DA redox cycle and proteinmetabolism. Pursuing the aim to untangle the intricatepathway of DA oxidation, we tested the peroxidative activitystaining in rat brain tissue homogenate, in order to make useof our method of investigation in PD animal models.Distinct rat brain areas were analysed: striatum, substantianigra, brain stem, frontal lobe, cerebellum. A red-orangeactivity gel band, similar to that previously developed fromhuman midbrain tissue (2), appeared in all the rat brainregions selected. Reactivity of striatum and substantia nigrawere expected, brain stem and frontal lobe showed sharp andfaint colour reaction, respectively. Remarkable enzymaticactivity was also developed from cerebellum. Massspectrometry analysis of the specific activity bands is nowin progress, in order to identify the proteins responsible ofthe enzymatic reaction. Present results appear to validate ourprocedure, providing an additional and useful approach tothe study of oxidative metabolism of DA.

Dopamine peroxidation: a novel study from rat brain areas

ARRIGONI, GIORGIO;GIUSTI, PIETRO;VIANELLO, FABIO;
2009

Abstract

Oxidative metabolism of dopamine (DA) appears to beinvolved in several, intersecting pathways underlyingParkinson’s disease (PD) pathogenesis (1). We have recentlyisolated an enzymatic activity from human midbrain tissue,by means of a specific in gel-staining procedure, set out byus, using DA and hydrogen peroxide as substrates (2). Massspectrometry analysis of the DA peroxidising activityrevealed the presence of proteins that may play a role inneurodegenerative disease, highlighting a possiblefunctional link among DA redox cycle and proteinmetabolism. Pursuing the aim to untangle the intricatepathway of DA oxidation, we tested the peroxidative activitystaining in rat brain tissue homogenate, in order to make useof our method of investigation in PD animal models.Distinct rat brain areas were analysed: striatum, substantianigra, brain stem, frontal lobe, cerebellum. A red-orangeactivity gel band, similar to that previously developed fromhuman midbrain tissue (2), appeared in all the rat brainregions selected. Reactivity of striatum and substantia nigrawere expected, brain stem and frontal lobe showed sharp andfaint colour reaction, respectively. Remarkable enzymaticactivity was also developed from cerebellum. Massspectrometry analysis of the specific activity bands is nowin progress, in order to identify the proteins responsible ofthe enzymatic reaction. Present results appear to validate ourprocedure, providing an additional and useful approach tothe study of oxidative metabolism of DA.
2009
Poster Session 2, Monday 14 September
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11577/2483304
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