Using FluoZin-3 and fura-2 to monitor acute accumulation of free intracellular Cd 2+ in a pancreatic beta cell line

The understanding of cellular Cd accumulation and toxicity is hampered by a lack of fluorescent indicators selective for intracellular free Cd ([Cd ] ). In this study, we used depolarized MIN6 mouse pancreatic beta cells as a model for evaluating [Cd ] detection with commercially available fluoresce...

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Bibliographic Details
Published in:Biometals 2019-12, Vol.32 (6), p.951
Main Authors: Malaiyandi, Latha M, Sharthiya, Harsh, Barakat, Ameir N, Edwards, Joshua R, Dineley, Kirk E
Format: Article
Language:English
Subjects:
Animals
Cadmium - analysis
Cadmium - metabolism
Cell Line
Fluorescent Dyes - analysis
Fluorescent Dyes - chemistry
Fura-2 - analysis
Fura-2 - chemistry
Insulin-Secreting Cells - chemistry
Insulin-Secreting Cells - metabolism
Mass Spectrometry
Mice
Microscopy, Fluorescence
Polycyclic Compounds - analysis
Polycyclic Compounds - chemistry
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Summary:The understanding of cellular Cd accumulation and toxicity is hampered by a lack of fluorescent indicators selective for intracellular free Cd ([Cd ] ). In this study, we used depolarized MIN6 mouse pancreatic beta cells as a model for evaluating [Cd ] detection with commercially available fluorescent probes, most of which have been traditionally used to visualize [Ca ] and [Zn ] . We trialed a panel of 12 probes including fura-2, FluoZin-3, Leadmium Green, Rhod-5N, indo-1, Fluo-5N, and others. We found that the [Zn ] probe FluoZin-3 and the traditional [Ca ] probe fura-2 responded most consistently and robustly to [Cd ] accumulation mediated by voltage-gated calcium channels. While selective detection of [Cd ] by fura-2 required the omission of Ca from extracellular buffers, FluoZin-3 responded to [Cd ] similarly in the presence or absence of extracellular Ca . Furthermore, we showed that FluoZin-3 and fura-2 can be used together for simultaneous monitoring of [Ca ] and [Cd ] in the same cells. None of the other fluorophores tested were effective [Cd ] detectors in this model.
ISSN:1572-8773