Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 450, Issue 4, Pages 1626-1630Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2014.07.049
Keywords
Fura-2-AM; Resveratrol; Fluorescence; Ca2+; HeLa cells; Interference
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Funding
- KU Leuven/Research Fund KU Leuven [DBOF/12/051]
- Onderzoeksfonds KU Leuven/Research Fund KU Leuven [OT/14/101]
- Interuniversity Attraction Poles Program (Belgian Science Policy) [P7/13]
- Research Foundation-Flanders (FWO) [G.0634.13, G.0819.13]
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Fura-2 is a commonly used fluorescent Ca2+ dye that allows an accurate determination of cytosolic Ca2+ levels by measuring the emission obtained at 510 nm after alternating excitation at 340 nm and 380 nm (F-340/F-330 ratio). Previous studies, based on Fura-2 measurements, claimed that resveratrol, a polyphenol implicated in human health, triggered an acute rise in cytosolic [Ca2+]. In this report, we show that the spectral properties of resveratrol are not compatible with the fluorescent properties of Fura-2. Resveratrol displays a strong absorption of light at a wavelength of 340 nm and a strong emission at 510 nm upon excitation at 340 nm (F-340). As a consequence, the F-340 values, but not the F-380 values, are increased when incubating cells with resveratrol. Consequently the F-340/(380) ratio values acutely increase upon addition of resveratrol, independently of changes in cytosolic [Ca2+]. Yet, we show that pretreating cells with resveratrol does not affect the F-340/F-380 ratios of Fura-2, provided that resveratrol is washed away before fluorescence measurement. These results indicate that Fura-2 is not suitable for assessing acute effects of resveratrol on Ca2+ signaling but that long-time effects can be assessed, provided that the resveratrol is carefully removed by appropriate wash steps. (C) 2014 Elsevier Inc. All rights reserved.
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