Abstract
Resveratrol, a naturally occurring polyphenol found in some fruits and especially in grapes, has been reported to provide diverse health benefits. Resveratrol’s mechanism of action is the subject of many investigations, and some studies using the ratiometric calcium indicator Fura-2 suggest that it modulates cellular calcium responses. In the current study, contradictory cellular calcium responses to resveratrol applied at concentrations exceeding 10 μM were observed during in vitro imaging studies depending on the calcium indicator used, with Fura-2 indicating an increase in intracellular calcium while Fluo-4 and the calcium biosensor YC3.60 indicated no response. When cells loaded with Fura-2 were treated with 100 μM resveratrol, excitation at 340 nm resulted in a large intensity increase at 510 nm, but the expected concurrent decline with 380 nm excitation was not observed. Pre-treatment of cells with the calcium chelator BAPTA-AM did not prevent a rise in the 340/380 ratio when resveratrol was present, but it did prevent an increase in 340/380 when ATP was applied, suggesting that the resveratrol response was an artifact. Cautious data interpretation is recommended from imaging experiments using Fura-2 concurrently with resveratrol in calcium imaging experiments.
References
Yu W, Fu Y-C, Wang W (2012) Cellular and molecular effects of resveratrol in health and disease. J Cell Biochem 113:752
Chung JH, Manganiello V, Dyck JRB (2012) Resveratrol as a calorie restriction mimetic: therapeutic implications. Trends Cell Biol 22:546
Park S, Ahmad F, Philp A, Baar K, Williams T, Luo H, Ke H, Rehmann H, Taussig R, Brown AL, Kim MK, Beaven MA, Burgin AB, Manganiello V, Chung JH (2012) Resveratrol ameliorates aging-related metabolic phenotypes by inhibiting cAMP phosphodiesterases. Cell 148:421
Jakab M, Lach S, Bacova Z, Langeluddecke C, Strbak V, Schmidt S, Iglseder E, Paulmichl M, Geibel J, Ritter M (2008) Resveratrol inhibits electrical activity and insulin release from insulinoma cells by block of voltage-gated Ca2+ channels and swelling-dependent Cl- currents. Cell Physiol Biochem 22:567
Dobrydneva Y, Williams RL, Blackmore PF (1999) trans-Resveratrol inhibits calcium influx in thrombin-stimulated human platelets. Br J Pharmacol 128:149
Dobrydneva Y, Williams RL, Blackmore PF (2010) Diethylstilbestrol and other nonsteroidal estrogens: novel class of store-operated calcium channel modulators. J Cardiovasc Pharmacol 55:522
Roe MW, Fiekers JF, Philipson LH, Bindokas VP (2006) Visualizing calcium signaling in cells by digitized wide-field and confocal fluorescent microscopy. Methods Mol Biol 319:37
Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440
Miyawaki A, Llopis J, Heim R, McCaffery JM, Adams JA, Ikura M, Tsien RY (1997) Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin. Nature 388:882
Palmer AE, Giacomello M, Kortemme T, Hires SA, Lev-Ram V, Baker D, Tsien RY (2006) Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs. Chem Biol 13:521
Paredes RM, Etzler JC, Watts LT, Lechleiter JD (2008) Chemical calcium indicators. Methods 46:143
Garcia-Sanchez L, Santofimia-Castano P, Miro-Moran A, Tapia JA, Salido GM, Gonzalez A (2012) Resveratrol mobilizes Ca2+ from intracellular stores and induces c-Jun N-terminal kinase activation in tumoral AR42J cells. Mol Cell Biochem 362:15
Campos-Toimil M, Elies J, Alvarez E, Verde I, Orallo F (2007) Effects of trans- and cis-resveratrol on Ca2+ handling in A7r5 vascular myocetes. Eur J Pharmacol 577:91
Sareen D, Darjatmoko SR, Albert DM, Polans AS (2007) Mitochondria, calcium, and calpain are key mediators of resveratrol-induced apoptosis in breast cancer. Mol Pharmacol 72:1466
Zhang J, Wu P, Long L, Chen Y, Hu Z, Ni L, Wang F, Chen J (2012) Resveratrol promotes cellular glucose utilization in primary cultured cortical neurons via calcium-dependent signaling pathway. J Nutr Biochem 24:629
Nagai T, Yamada S, Tominaga T, Ichikawa M, Miyawaki A (2004) Expanded dynamic range of fluorescent indicators for Ca2+ by circularly permuted yellow fluorescent proteins. PNAS 101:10554
Landa LR, Harbeck M, Kaihara K, Chepurny O, Kitiphongspattana J, Graf O, Nikolaev VO, Lohse MJ, Holz GG, Roe MW (2005) Interplay of Ca2+ and cAMP signaling in the insulin-secreting MIN6 β-cell line. J Biol Chem 280:31294
Gee KR, Brown KA, Chen WU, Bishop-Stewart J, Gray JD, Johnson I (2000) Chemical and physiological characterization of fluo-4 Ca2+- indicator dyes. Cell Calcium 27:97
Qian C, Ma J, Zhang P, Luo A, Wang C, Ren Z, Kong L, Zhang S, Wang X, Wu Y (2012) Resveratrol attenuates the Na+-dependent intracellular Ca2+ overload by inhibiting H2O2 induced increase in late sodium current in ventricular myocytes. PLOS ONE 7(12):e51358
Elies J, Cuinas A, Garcia-Morales V, Orallo F, Campos-Toimil M (2011) Trans-resveratrol simultaneously increases cytoplasmic Ca2+ levels and nitric oxide release in human endothelial cells. Mol Nutr Food Res 55:1237
Acknowledgments
This work was supported by a Basic Science Award from the American Diabetes Association to CAL and NIH grants R01 DK074966 and R01 DK092616 to MWR. Adenovirus YC3.60 was a generous gift from Dr. Christopher Rhodes, University of Chicago, Chicago, IL.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kopp, R.F., Leech, C.A. & Roe, M.W. Resveratrol Interferes with Fura-2 Intracellular Calcium Measurements. J Fluoresc 24, 279–284 (2014). https://doi.org/10.1007/s10895-013-1312-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10895-013-1312-9