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Journal of Histochemistry and Cytochemistry, Vol. 50, 731-734, May 2002, Copyright © 2002, The Histochemical Society, Inc.

BRIEF REPORT

The Fluorescent Probe Bodipy-FL–Verapamil Is a Substrate for Both P-glycoprotein and Multidrug Resistance-related Protein (MRP)-1

Enrico Crivellatoa, Luigi Candussiob, Anna M. Rosatib, Fiora Bartoli–Klugmannb, Franco Mallardia, and Giuliana Decortib
a Department of Medical and Morphological Researches, Anatomy Section, University of Udine, Udine, Italy
b Department of Biomedical Sciences, University of Trieste, Trieste, Italy

Correspondence to: Enrico Crivellato, Dept. of Medical and Morphological Researches, Anatomy Section, P.le Kolbe, n.3, 33100 Udine, Italy. E-mail: enrico.crivellato@drmm.uniud.it

Several fluorescent probes have been used in functional studies to analyze drug transport in multidrug-resistant cells by fluorescent microscopy. Because many of these molecules have some drawbacks, such as toxicity, nonspecific background, or accumulation in mitochondria, new fluorescent compounds have been proposed as more useful tools. Among these substances, Bodipy-FL–Verapamil, a fluorescent conjugate of the drug efflux blocker verapamil, has been used to study P-glycoprotein activity in different cell types. In this study we tested by fluorescent microscopy the accumulation of Bodipy-FL–Verapamil in cell lines that overexpress either P-glycoprotein (P-gp) or multidrug resistance-related protein 1 (MRP1). Expression of P-gp and MRP1 was evaluated at the mRNA level by RT-PCR technique and at the protein level by flow cytometric analysis using C219 and MRP-m6 monoclonal antibodies. Results indicate that Bodipy-FL–Verapamil is actually a substrate for both proteins. As a consequence, any conclusion about P-gp activity obtained by the use of Bodipy-FL–Verapamil as fluorescent tracer should be interpreted with caution. (J Histochem Cytochem 50:731–734, 2002)

Key Words: Bodipy-FL–Verapamil, P-glycoprotein, multidrug resistance-related, Protein 1, fluorescence microscopy


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