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Title
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A rapid method for the evaluation of compounds with mitochondria-protective properties
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Author
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Abstract
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| 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. |
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Language
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English
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Source (journal)
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Journal of neuroscience methods. - Amsterdam
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Publication
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Amsterdam
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1999
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ISSN
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0165-0270
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DOI
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10.1016/S0165-0270(99)00107-7
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Volume/pages
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92
:1-2
(1999)
, p. 153-159
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ISI
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000084042800017
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Full text (Publisher's DOI)
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