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    Comparative Ca 2+ channel contributions to intracellular Ca 2+ levels in the circadian clock

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    Author
    Plante, Amber E
    Rao, Vishnu P
    Rizzo, Megan A
    Meredith, Andrea L
    Date
    2021-07-21
    Journal
    Biophysical Reports
    Publisher
    Elsevier
    Type
    Article
    
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    See at
    https://doi.org/10.1016/j.bpr.2021.100005
    http://www.ncbi.nlm.nih.gov/pmc/articles/pmc8942421/
    Abstract
    Circadian rhythms in mammals are coordinated by the central clock in the brain, located in the suprachiasmatic nucleus (SCN). Multiple molecular and cellular signals display a circadian variation within SCN neurons, including intracellular Ca2+, but the mechanisms are not definitively established. SCN cytosolic Ca2+ levels exhibit a peak during the day, when both action potential firing and Ca2+ channel activity are increased, and are decreased at night, correlating with a reduction in firing rate. In this study, we employ a single-color fluorescence anisotropy reporter (FLARE), Venus FLARE-Cameleon, and polarization inverted selective-plane illumination microscopy to measure rhythmic changes in cytosolic Ca2+ in SCN neurons. Using this technique, the Ca2+ channel subtypes contributing to intracellular Ca2+ at the peak and trough of the circadian cycle were assessed using a pharmacological approach with Ca2+ channel inhibitors. Peak (218 ± 16 nM) and trough (172 ± 13 nM) Ca2+ levels were quantified, indicating a 1.3-fold circadian variance in Ca2+ concentration. Inhibition of ryanodine-receptor-mediated Ca2+ release produced a larger relative decrease in cytosolic Ca2+ at both time points compared to voltage-gated Ca2+channels. These results support the hypothesis that circadian Ca2+ rhythms in SCN neurons are predominantly driven by intracellular Ca2+ channels, although not exclusively so. The study provides a foundation for future experiments to probe Ca2+ signaling in a dynamic biological context using FLAREs.
    Identifier to cite or link to this item
    http://hdl.handle.net/10713/18373
    ae974a485f413a2113503eed53cd6c53
    10.1016/j.bpr.2021.100005
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