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

• Published in: International journal of molecular sciences Vol. 13; no. 8; pp. 9615 - 9626
• Main Authors: Pogue, Aileen I, Jones, Brandon M, Bhattacharjee, Surjyadipta, Percy, Maire E, Zhao, Yuhai, Lukiw, Walter J
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.08.2012; Molecular Diversity Preservation International (MDPI)
• Subjects: 5-carboxy-2′,7′-dichlorofluorescein diacetate; 6-carboxy-2′,7′-dichlorofluorescein diacetate (5- and 6-carboxy-DCFDA; Aging; Aluminum; Alzheimer's disease; Amyotrophic lateral sclerosis; Brain; Brain - drug effects; Brain - metabolism; Brain - pathology; Brief Report; carboxy-DCFDA; Cell Membrane Permeability; Cell Tracking; Cells, Cultured; Disease; epigenetic human neural cells; Epigenetics; Fluoresceins; Fluorescent Dyes; Free radicals; Humans; inflammation; Metabolism; metal sulfates; Metals - chemistry; Metals - pharmacology; Molecular structure; Neurological disorders; Neurons - drug effects; Neurons - metabolism; Neurons - pathology; Oxidative stress; Oxidative Stress - drug effects; Parkinson's disease; prion disease; Reactive Oxygen Species - metabolism; Sulfates - chemistry; Sulfates - pharmacology; synergistic effects; Canada; United States > US; epigenetic human neural cells; 5-carboxy-2′,7′-dichlorofluorescein diacetate; metal sulfates; prion disease; Alzheimer’s disease; inflammation; synergistic effects; 6-carboxy-2′,7′-dichlorofluorescein diacetate (5- and 6-carboxy-DCFDA; carboxy-DCFDA); carboxy-DCFDA; aluminum; amyotrophic lateral sclerosis; Parkinson’s disease
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms13089615

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.