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Research Article Free access | 10.1172/JCI116781

Regulation of Na,K-adenosine triphosphatase gene expression by sodium ions in cultured neonatal rat cardiocytes.

K Yamamoto, U Ikeda, Y Seino, Y Tsuruya, A Oguchi, K Okada, S Ishikawa, T Saito, K Kawakami, and Y Hara

Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Department of Cardiology, Jichi Medical School, Tochigi, Japan.

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Published October 1, 1993 - More info

Published in Volume 92, Issue 4 on October 1, 1993
J Clin Invest. 1993;92(4):1889–1895. https://doi.org/10.1172/JCI116781.
© 1993 The American Society for Clinical Investigation
Published October 1, 1993 - Version history
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Abstract

Na,K-ATPase (Na,K-pump) plays an important role in the regulation of intracellular ion composition. The purpose of this study is to determine whether Na+ regulates the levels of mRNA coding for Na,K-ATPase alpha and beta subunits in cultured neonatal rat cardiocytes. We measured intracellular Na+ levels ([Na+]i) in cardiocytes using a Na(+)-sensitive fluorescence dye (SBFI). 1 mM ouabain caused a significant increase in [Na+]i in cardiocytes; from 12.8 +/- 0.3 to 28.8 +/- 1.8 mM. Exposure of cardiocytes to 1 mM ouabain resulted in a three- to fourfold increase in alpha 1, alpha 2, and alpha 3 mRNA accumulation, and an approximate two-fold increase in beta 1 mRNA accumulation. A maximum elevation was reached at 60 min in both cases. The ouabain-induced alpha 1 mRNA accumulation was still observed in the Ca(2+)-free culture medium. Exposure of cardiocytes to 10 microM monensin in the absence of extracellular Ca2+ also resulted in a threefold increase in alpha 1 mRNA accumulation. The increased alpha 1 mRNA expression by 1 mM ouabain was associated with a fourfold increase in alpha 1 subunit protein accumulation. Transfection experiments with chimeric plasmids containing 5'-flanking sequences of alpha 1, alpha 2, and alpha 3 isoform genes and a luciferase reporter gene revealed that 1 mM ouabain caused a twofold increase in luciferase activity in each alpha system. These results suggest that Na+ directly regulates Na,K-ATPase gene expression in cardiocytes. The transfection study further supports the premise that Na(+)-responsive elements are located within the 5'-flanking sequences of each alpha isoform gene.

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