Transcriptomic changes triggered by ouabain in rat cerebellum granule cells: Role of α3- and α1-Na+,K+-ATPase-mediated signaling

PLoS One. 2019 Sep 26;14(9):e0222767. doi: 10.1371/journal.pone.0222767. eCollection 2019.

Abstract

It was shown previously that inhibition of the ubiquitous α1 isoform of Na+,K+-ATPase by ouabain sharply affects gene expression profile via elevation of intracellular [Na+]i/[K+]i ratio. Unlike other cells, neurons are abundant in the α3 isoform of Na+,K+-ATPase, whose affinity in rodents to ouabain is 104-fold higher compared to the α1 isoform. With these sharp differences in mind, we compared transcriptomic changes in rat cerebellum granule cells triggered by inhibition of α1- and α3-Na+,K+-ATPase isoforms. Inhibition of α1- and α3-Na+,K+-ATPase isoforms by 1 mM ouabain resulted in dissipation of transmembrane Na+ and K+ gradients and differential expression of 994 transcripts, whereas selective inhibition of α3-Na+,K+-ATPase isoform by 100 nM ouabain affected expression of 144 transcripts without any impact on the [Na+]i/[K+]i ratio. The list of genes whose expression was affected by 1 mM ouabain by more than 2-fold was abundant in intermediates of intracellular signaling and transcription regulators, including augmented content of Npas4, Fos, Junb, Atf3, and Klf4 mRNAs, whose upregulated expression was demonstrated in neurons subjected to electrical and glutamatergic stimulation. The role [Na+]i/[K+]i-mediated signaling in transcriptomic changes involved in memory formation and storage should be examined further.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cardiotonic Agents / pharmacology*
  • Cells, Cultured
  • Cerebellum / cytology
  • Cerebellum / drug effects*
  • Cerebellum / metabolism
  • Cytoplasmic Granules / drug effects
  • Cytoplasmic Granules / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation / drug effects*
  • Isoenzymes / genetics
  • Isoenzymes / metabolism
  • Kruppel-Like Factor 4
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / metabolism
  • Ouabain / pharmacology*
  • Primary Cell Culture
  • Rats
  • Signal Transduction / drug effects
  • Signal Transduction / genetics
  • Sodium-Potassium-Exchanging ATPase / genetics
  • Sodium-Potassium-Exchanging ATPase / metabolism*
  • Transcription, Genetic / drug effects
  • Transcriptome / drug effects

Substances

  • Cardiotonic Agents
  • Isoenzymes
  • Klf4 protein, rat
  • Kruppel-Like Factor 4
  • Ouabain
  • Sodium-Potassium-Exchanging ATPase

Grants and funding

This work was supported by grants from the Russian Scientific Foundation RNF http://rscf.ru/ (#16-15-10026-п) (S.N.O) and the Russian Foundation for Basic Research RFBR http://www.rfbr.ru/rffi/ru/ (#18-04-00063) (S.N.O). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.