title: Detection of Free and Protein-Bound ortho-Quinones by Near-Infrared Fluorescence.
abstract: Aging and oxidative stress are two prominent pathological mechanisms for Parkinson's disease (PD) that are strongly associated with the degeneration of dopamine (DA) neurons in the midbrain. DA and other catechols readily oxidize into highly reactive o-quinone species that are precursors of neuromelanin (NM) pigment and under pathological conditions can modify and damage macromolecules. The role of DA oxidation in PD pathogenesis remains unclear in part due to the lack of appropriate disease models and the absence of a simple method for the quantification of DA-derived oxidants. Here, we describe a rapid, simple, and reproducible method for the quantification of o-quinones in cells and tissues that relies on the near-infrared fluorescent properties of these species. Importantly, we demonstrate that catechol-derived oxidants can be quantified in human neuroblastoma cells and midbrain dopamine neurons derived from induced pluripotent stem cells, providing a novel model to study the downstream actions of o-quinones. This method should facilitate further study of oxidative stress and DA oxidation in PD and related diseases that affect the dopaminergic system.
authors.affiliation: The Ken and Ruth Davee Department of Neurology, Northwestern University Feinberg School of Medicine , 303 East Chicago Avenue, Ward 12-144, Chicago Illinois 60611 United States.
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Mazzulli
authors.affiliation: The Ken and Ruth Davee Department of Neurology, Northwestern University Feinberg School of Medicine , 303 East Chicago Avenue, Ward 12-144, Chicago Illinois 60611 United States.
authors.forename: Lena F
authors.initials: LF
authors.lastname: Burbulla
authors.affiliation: The Ken and Ruth Davee Department of Neurology, Northwestern University Feinberg School of Medicine , 303 East Chicago Avenue, Ward 12-144, Chicago Illinois 60611 United States.
authors.forename: Dimitri
authors.initials: D
authors.lastname: Krainc
authors.affiliation: Department of Pediatrics, Children's Hospital of Philadelphia , Philadelphia, Pennsylvania 19104, United States.|Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
authors.forename: Harry
authors.initials: H
authors.lastname: Ischiropoulos
mesh_terms: D002396:Catechols / Q000737:chemistry; D004298:Dopamine / Q000737:chemistry / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000737:chemistry* / Q000166:cytology; D005453:Fluorescence; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D007259:Infrared Rays; D008636:Mesencephalon / Q000166:cytology; D009447:Neuroblastoma / Q000737:chemistry* / Q000473:pathology; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology; D011809:Quinones / Q000032:analysis* / Q000737:chemistry*
chemical_list: D002396:Catechols; D011809:Quinones; C034221:catechol; D004298:Dopamine
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