Abstract—
In several types of cells, inhibition of ubiquitous α1-isoform of the Na+,K+-ATPase leads to drastic transcriptomic changes mediated by dissipation of transmembrane concentration gradients of monovalent cations. In the present study, we employed the sharp differences in the affinity of α1- and α3-Na+,K+-ATPase for ouabain to examine the role of α3 isoform in the regulation of intracellular Na+ and K+ content and gene expression in primary cultures of rat cerebellum granule cells. Addition of 100 nM ouabain decreased the Na+,K+-ATPase activity by 20% due to the inhibition of α3 isosyme. At this concentration, ouabain changed transcription of 17 genes with maximal ~2-fold activation and 1.5-fold inhibition. The full-scale inhibition of α1- and α3-Na+,K+-ATPase by 1 mM ouabain was accompanied by a ~50-fold elevation of the [Na+]i/[K+]i ratio and altered the content of mRNA encoding 673 genes with maximal 20-fold activation and 3-fold inhibition. Unlike 1 mM ouabain, 100 nM ouabain did not affect phosphorylation of the Ca2+-sensitive transcription regulator CREB. Our results show that transcriptomic changes in neurons subjected to inhibition of α3-Na+,K+-ATPase by low doses of ouabain are not mediated by elevation of the [Na+]i/[K+]i, ratio and activation-sensitive mechanisms of excitation–transcription coupling.
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ACKNOWLEDGMENTS
The work was supported by the Russian Scientific Foundation (project no. 16-15-10026) and the Russian Foundation for Basic Research (project no. 18-04-00063).
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All procedures were performed in accordance with the European Communities Council Directive (November 24, 1986; 86/609/EEC) and the Declaration on humane treatment of animals. The Protocol of experiments was approved by the bioethics committee of the Faculty of Biology, Lomonosov Moscow State University (no. 82-O).
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Smolyaninova, L.V., Shiyan, A.A., Klementieva, T.S. et al. Ouabain at Low Concentrations Affects Transcription without Any Impact on Intracellular Content of Sodium and Potassium in Rat Brain Neurons. Biochem. Moscow Suppl. Ser. A 13, 365–371 (2019). https://doi.org/10.1134/S1990747819030073
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DOI: https://doi.org/10.1134/S1990747819030073