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Int. J. Mol. Sci. 2012, 13(8), 9615-9626; doi:10.3390/ijms13089615
Article

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

1
, 2
, 2, 3
, 4
 and 2,*
1 Alchem Biotek Corporation, Toronto, ON M5T 1L8, Canada 2 LSU Neuroscience Center and Department of Ophthalmology, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA 3 Surrey Place Centre, University of Toronto, Toronto, ON M5S 1A8, Canada 4 University of Texas Health Science Center, Houston, TX 77030, USA
* Author to whom correspondence should be addressed.
Received: 21 June 2012 / Revised: 20 July 2012 / Accepted: 24 July 2012 / Published: 2 August 2012
(This article belongs to the Special Issue Advances in Free Radicals in Biology and Medicine)
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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; C25H14Cl2O9; 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-H2DCFDA 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.
Keywords: 5-carboxy-2′,7′-dichlorofluorescein diacetate; 6-carboxy-2′,7′-dichlorofluorescein diacetate (5- and 6-carboxy-DCFDA; carboxy-DCFDA); aluminum; Alzheimer’s disease; amyotrophic lateral sclerosis; carboxy-DCFDA; epigenetic human neural cells; inflammation; metal sulfates; synergistic effects; Parkinson’s disease; prion disease 5-carboxy-2′,7′-dichlorofluorescein diacetate; 6-carboxy-2′,7′-dichlorofluorescein diacetate (5- and 6-carboxy-DCFDA; carboxy-DCFDA); aluminum; Alzheimer’s disease; amyotrophic lateral sclerosis; carboxy-DCFDA; epigenetic human neural cells; inflammation; metal sulfates; synergistic effects; Parkinson’s disease; prion disease
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Pogue, A.I.; Jones, B.M.; Bhattacharjee, S.; Percy, M.E.; Zhao, Y.; Lukiw, W.J. 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. Int. J. Mol. Sci. 2012, 13, 9615-9626.

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