Title
A rapid method for the evaluation of compounds with mitochondria-protective propertiesA rapid method for the evaluation of compounds with mitochondria-protective properties
Author
Faculty/Department
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences . Biomedical Sciences
Research group
Department of Biomedical Sciences
Publication type
article
Publication
Amsterdam,
Subject
Chemistry
Biology
Human medicine
Source (journal)
Journal of neuroscience methods. - Amsterdam
Volume/pages
92(1999):1-2, p. 153-159
ISSN
0165-0270
ISI
000084042800017
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Mitochondrial dysfunction has been implicated in a number of neurodegenerative diseases, such as ischemia and Parkinson's disease. We present here a method that allows the rapid quantification of interventions, aimed at inhibiting the effect of mitochondrial membrane potential uncouplers, based on the ratioing properties of the fluorescent probe 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1), by using currently available 96-well fluorescent plate readers. A method is presented for evaluation of cross-talk between the two excitation/emission channels. Further characterization of the probe shows that the effect of plasma membrane potential changes on JC-I fluorescence ratio are negligible, but that the signal is very sensitive to pH. One of the most exciting applications is the possibility to perform end-point measurements, thanks to the ratioing properties of the probe. The system is tested in different culture types with different mitochondrial uncouplers. As an example of a quantitative evaluation, we show that flunarizine is able to inhibit, dose-dependently, FCCP mediated JC-l signal increase. The procedure is simple and allows for the fast screening of mitochondria-protective compounds (C) 1999 Elsevier Science B.V. All rights reserved.
E-info
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