Probenicid inhibition of fluorescence extrusion after MCB-staining of rat-1 fibroblasts |
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Authors: | Poot M Hudson F N Grossmann A Rabinovitch P S Kavanagh T J |
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Affiliation: | Department of Human Genetics, University of Würzburg, 97074 Würzburg, Germany. |
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Abstract: | The intracellular fluorescence level of cells stained continuously with monochlorobimane was monitored by flow cytometry in order to assess the initial rate of glutatione to monochlorobimane conjugation as a measure of glutathione S-transferase activity. In addition to a rapid initial increase and a plateau level, a decline in fluorescence intensity was found upon prolonged flow cytometric monitoring. Exposure to probenicid, an inhibitor of an ATP-dependent organic anion pump, prevented this decrease. Incubation with vanadate and verapamil was without effect. Thus, extrusion of fluorescentglutathione-conjugate perturbs the proportionality between initial glutathione level and monochlorobimane-dependent fluorescence intensity. Monitoring by flow cytometry the decrease in monochlorobimane-dependent fluorescence may be useful to detect multidrug resistant cells. |
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