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Fluorescent Ca2+ indicators directly inhibit the Na,K-ATPase and disrupt cellular functions.

ABSTRACT: Fluorescent Ca2+ indicators have been essential for the analysis of Ca2+ signaling events in various cell types. We showed that chemical Ca2+ indicators, but not a genetically encoded Ca2+ indicator, potently suppressed the activity of Na+- and K+-dependent adenosine triphosphatase (Na,K-ATPase), independently of their Ca2+ chelating activity. Loading of commonly used Ca2+ indicators, including Fluo-4 acetoxymethyl (AM), Rhod-2 AM, and Fura-2 AM, and of the Ca2+ 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%. Ca2+ 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 Ca2+ 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 Ca2+ indicators may be necessary.

SUBMITTER: Smith NA 

PROVIDER: S-EPMC6190706 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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Fluorescent Ca<sup>2+</sup> indicators directly inhibit the Na,K-ATPase and disrupt cellular functions.

Smith Nathan A NA   Kress Benjamin T BT   Lu Yuan Y   Chandler-Militello Devin D   Benraiss Abdellatif A   Nedergaard Maiken M  

Science signaling 20180130 515

Fluorescent Ca<sup>2+</sup> indicators have been essential for the analysis of Ca<sup>2+</sup> signaling events in various cell types. We showed that chemical Ca<sup>2+</sup> indicators, but not a genetically encoded Ca<sup>2+</sup> indicator, potently suppressed the activity of Na<sup>+</sup>- and K<sup>+</sup>-dependent adenosine triphosphatase (Na,K-ATPase), independently of their Ca<sup>2+</sup> chelating activity. Loading of commonly used Ca<sup>2+</sup> indicators, including Fluo-4 acetoxy  ...[more]

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