Fluorescent Ca indicators have been essential for the analysis of Ca signaling events in various cell types. We showed that chemical Ca indicators, but not a genetically encoded Ca indicator, potently suppressed the activity of Na- and K-dependent adenosine triphosphatase (Na,K-ATPase), independently of their Ca chelating activity. Loading of commonly used Ca indicators, including Fluo-4 acetoxymethyl (AM), Rhod-2 AM, and Fura-2 AM, and of the Ca chelator BAPTA AM into cultured mouse or human neurons, astrocytes, cardiomyocytes, or kidney proximal tubule epithelial cells suppressed Na,K-ATPase activity by 30 to 80%. Ca indicators also suppressed the agonist-induced activation of the Na,K-ATPase, altered metabolic status, and caused a dose-dependent loss of cell viability. Loading of Ca indicators into mice, which is carried out for two-photon imaging, markedly altered brain extracellular concentrations of K and ATP. These results suggest that a critical review of data obtained with chemical Ca indicators may be necessary.
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| http://dx.doi.org/10.1126/scisignal.aal2039 | DOI Listing |
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