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Na,K-ATPase Activity in Mouse Muscle is Regulated by AMPK and PGC-1α

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Abstract

Na,K-ATPase activity, which is crucial for skeletal muscle function, undergoes acute and long-term regulation in response to muscle activity. The aim of the present study was to test the hypothesis that AMP kinase (AMPK) and the transcriptional coactivator PGC-1α are underlying factors in long-term regulation of Na,K-ATPase isoform (α,β and PLM) abundance and Na+ affinity. Repeated treatment of mice with the AMPK activator AICAR decreased total PLM protein content but increased PLM phosphorylation, whereas the number of α- and β-subunits remained unchanged. The K m for Na+ stimulation of Na,K-ATPase was reduced (higher affinity) after AICAR treatment. PLM abundance was increased in AMPK kinase-dead mice compared with control mice, but PLM phosphorylation and Na,K-ATPase Na+ affinity remained unchanged. Na,K-ATPase activity and subunit distribution were also measured in mice with different degrees of PGC-1α expression. Protein abundances of α1 and α2 were reduced in PGC-1α +/− and −/− mice, and the β1/β2 ratio was increased with PGC-1α overexpression (TG mice). PLM protein abundance was decreased in TG mice, but phosphorylation status was unchanged. Na,K-ATPase V max was decreased in PCG-1α TG and KO mice. Experimentally in vitro induced phosphorylation of PLM increased Na,K-ATPase Na+ affinity, confirming that PLM phosphorylation is important for Na,K-ATPase function. In conclusion, both AMPK and PGC-1α regulate PLM abundance, AMPK regulates PLM phosphorylation and PGC-1α expression influences Na,K-ATPase α1 and α2 content and β1/β2 isoform ratio. Phosphorylation of the Na,K-ATPase subunit PLM is an important regulatory mechanism.

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Acknowledgments

The present study was supported by the Carlsberg Foundation. We thank Prof. B. Spiegelman for providing PGC-1α mice and M. J. Birnbaum for providing KD mice initially to start breeding in house. We also thank Joachim Fentz, Lotte Leick and Jonas T. Treebak for help handling animals and Helle Walas for skilled technical assistance.

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Authors and Affiliations

  1. Department of Biology, University of Copenhagen, August Krogh Building, Universitetsparken 13, Copenhagen, 2100, Denmark

    Maria S. Ingwersen, Michael Kristensen, Henriette Pilegaard & Carsten Juel

  2. Institute of Exercise and Sport Sciences, University of Copenhagen, Copenhagen, Denmark

    Jørgen F. P. Wojtaszewski & Erik A. Richter

Authors
  1. Maria S. Ingwersen
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  2. Michael Kristensen
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  3. Henriette Pilegaard
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  4. Jørgen F. P. Wojtaszewski
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  5. Erik A. Richter
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  6. Carsten Juel
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Correspondence to Carsten Juel.

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Ingwersen, M.S., Kristensen, M., Pilegaard, H. et al. Na,K-ATPase Activity in Mouse Muscle is Regulated by AMPK and PGC-1α. J Membrane Biol 242, 1–10 (2011). https://doi.org/10.1007/s00232-011-9365-7

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  • DOI: https://doi.org/10.1007/s00232-011-9365-7

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