title: Comparison of fluorescence probes for intracellular sodium imaging in prostate cancer cell lines.
abstract: Sodium (Na+) ions are known to regulate many signaling pathways involved in both physiological and pathological conditions. In particular, alterations in intracellular concentrations of Na+and corresponding changes in membrane potential are known to be major actors of cancer progression to metastatic phenotype. Though the functionality of Na+channels and the corresponding Na+currents can be investigated using the patch-clamp technique, the latter is rather invasive and a technically difficult method to study intracellular Na+transients compared to Na+fluorescence imaging. Despite the fact that Na+signaling is considered an important controller of cancer progression, only few data using Na+imaging approaches are available so far, suggesting the persisting challenge within the scientific community. In this study, we describe in detail the approach for application of Na+imaging technique to measure intracellular Na+variations in human prostate cancer cells. Accordingly, we used three Na+-specific fluorescent dyes-Na+-binding benzofuran isophthalate (SBFI), CoroNa™ Green (Corona) and Asante NaTRIUM Green-2 (ANG-2). These dyes have been assessed for optimal loading conditions, dissociation constant and working range after different calibration methods, and intracellular Na+sensitivity, in order to determine which probe can be considered as the most reliable to visualize Na+fluctuations in vitro.
authors.affiliation: Inserm U1003, Laboratory of Excellence, Ion Channels Science and Therapeutics, Equipe Labellisée par la Ligue Nationale Contre le Cancer, SIRIC ONCOLille, Université des Sciences et Technologies de Lille, 59656, Villeneuve d'Ascq, France.
authors.forename: Oksana
authors.identifier: http://orcid.org/0000-0002-1984-0013
authors.initials: O
authors.lastname: Iamshanova
authors.affiliation: Inserm U1003, Laboratory of Excellence, Ion Channels Science and Therapeutics, Equipe Labellisée par la Ligue Nationale Contre le Cancer, SIRIC ONCOLille, Université des Sciences et Technologies de Lille, 59656, Villeneuve d'Ascq, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Mariot
authors.affiliation: Inserm U1003, Laboratory of Excellence, Ion Channels Science and Therapeutics, Equipe Labellisée par la Ligue Nationale Contre le Cancer, SIRIC ONCOLille, Université des Sciences et Technologies de Lille, 59656, Villeneuve d'Ascq, France.
authors.forename: V'yacheslav
authors.initials: V
authors.lastname: Lehen'kyi
authors.affiliation: Inserm U1003, Laboratory of Excellence, Ion Channels Science and Therapeutics, Equipe Labellisée par la Ligue Nationale Contre le Cancer, SIRIC ONCOLille, Université des Sciences et Technologies de Lille, 59656, Villeneuve d'Ascq, France. natacha.prevarskaya@univ-lille1.fr.
authors.forename: Natalia
authors.initials: N
authors.lastname: Prevarskaya
mesh_terms: D002138:Calibration; D045744:Cell Line, Tumor; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism*; D015640:Ion Channel Gating / Q000187:drug effects; D008297:Male; D057054:Molecular Imaging / Q000379:methods*; D011471:Prostatic Neoplasms / Q000473:pathology*; D012964:Sodium / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D061566:Voltage-Gated Sodium Channels / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D061566:Voltage-Gated Sodium Channels; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase
keywords: ANG-2; CoroNa Green; Fluorescent dye; Prostate cancer cells; SBFI; Sodium imaging