title: Metal-sulfate induced generation of ROS in human brain cells: detection using an isomeric mixture of 5- and 6-carboxy-2',7'-dichlorofluorescein diacetate (carboxy-DCFDA) as a cell permeant tracer.
abstract: Evolution of reactive oxygen species (ROS), generated during the patho-physiological stress of nervous tissue, has been implicated in the etiology of several progressive human neurological disorders including Alzheimer's disease (AD) and amylotrophic lateral sclerosis (ALS). In this brief communication we used mixed isomers of 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate (carboxy-DCFDA; C(25)H(14)C(l2)O(9); MW 529.3), a novel fluorescent indicator, to assess ROS generation within human neuronal-glial (HNG) cells in primary co-culture. We introduced pathological stress using the sulfates of 12 environmentally-, industrially- and agriculturally-relevant divalent and trivalent metals including Al, Cd, Cu, Fe, Hg, Ga, Mg, Mn, Ni, Pb, Sn and Zn. In this experimental test system, of all the metal sulfates analyzed, aluminum sulfate showed by far the greatest ability to induce intracellular ROS. These studies indicate the utility of using isomeric mixtures of carboxy-H(2)DCFDA diacetates as novel and highly sensitive, long-lasting, cell-permeant, fluorescein-based tracers for quantifying ROS generation in intact, metabolizing human brain cells, and in analyzing the potential epigenetic contribution of different metal sulfates to ROS-generation and ROS-mediated neurological dysfunction.
authors.affiliation: Alchem Biotek Corporation, Toronto, ON M5T 1L8, Canada.
authors.forename: Aileen I
authors.initials: AI
authors.lastname: Pogue
authors.affiliation: LSU Neuroscience Center and Department of Ophthalmology, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA.
authors.forename: Brandon M
authors.initials: BM
authors.lastname: Jones
authors.affiliation: LSU Neuroscience Center and Department of Ophthalmology, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA.
authors.forename: Surjyadipta
authors.initials: S
authors.lastname: Bhattacharjee
authors.affiliation: Surrey Place Centre, University of Toronto, Toronto, ON M5S 1A8, Canada.
authors.forename: Maire E
authors.initials: ME
authors.lastname: Percy
authors.affiliation: University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Yuhai
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: LSU Neuroscience Center and Department of Ophthalmology, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA.
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Lukiw
mesh_terms: D001921:Brain / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D002463:Cell Membrane Permeability; D058948:Cell Tracking; D002478:Cells, Cultured; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006801:Humans; D008670:Metals / Q000737:chemistry / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013431:Sulfates / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D005452:Fluoresceins; D005456:Fluorescent Dyes; D008670:Metals; D017382:Reactive Oxygen Species; D013431:Sulfates; C053897:5(6)-carboxy-2',7'-dichlorofluorescein