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Functional Interaction Between Na/K-ATPase and NMDA Receptor in Cerebellar Neurons

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Abstract

NMDA receptors play a crucial role in regulating synaptic plasticity and memory. Activation of NMDA receptors changes intracellular concentrations of Na+ and K+, which are subsequently restored by Na/K-ATPase. We used immunochemical and biochemical methods to elucidate the potential mechanisms of interaction between these two proteins. We observed that NMDA receptor and Na/K-ATPase interact with each other and this interaction was shown for both isoforms of α subunit (α1 and α3) of Na/K-ATPase expressed in neurons. Using Western blotting, we showed that long-term exposure of the primary culture of cerebellar neurons to nanomolar concentrations of ouabain (a cardiotonic steroid, a specific ligand of Na/K-ATPase) leads to a decrease in the levels of NMDA receptors which is likely mediated by the α3 subunit of Na/K-ATPase. We also observed a decrease in enzymatic activity of the α1 subunit of Na/K-ATPase caused by NMDA receptor activation. This effect is mediated by an increase in intracellular Ca2+. Thus, Na/K-ATPase and NMDA receptor can interact functionally by forming a macromolecular complex which can be important for restoring ionic balance after neuronal excitation. Furthermore, this interaction suggests that NMDA receptor function can be regulated by endogenous cardiotonic steroids which recently have been found in cerebrospinal fluid or by pharmacological drugs affecting Na/K-ATPase function.

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Acknowledgments

We dedicate this article to the memory of Professor Alexander Boldyrev, who passed away in 2012. Professor Boldyrev worked for more than 40 years in this field and created the Moscow school of Na/K-ATPase research at Moscow State University. We thank Professor Amir Askari for the opportunity to perform a part of this work in his Laboratory. We also thank Professor Anita Aperia for the opportunity to do a part of this work in her Laboratory and for her comments on the manuscript. This work was supported by RFBR grants 12-04-31468 and 14-04-32123, the Russian President Grant for 2010/2011, and NIH HL036573. We also received financial support from internal postdoctoral grant from St. Petersburg State University.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Faculty of Biology and Soil Sciences, St. Petersburg State University, St. Petersburg, Russia

    Evgeny E. Akkuratov

  2. Department of Biochemistry and Cancer Biology, College of Medicine and Life Sciences, Health Science Campus, University of Toledo, Toledo, USA

    Evgeny E. Akkuratov & Lijun Liu

  3. Research Center of Neurology Russian Academy of Medical Sciences, Moscow, Russia

    Olga M. Lopacheva, Alexandr V. Lopachev & Alexander A. Boldyrev

  4. International Biotechnological Center, Lomonosov Moscow State University, Moscow, Russia

    Olga M. Lopacheva & Alexander A. Boldyrev

  5. Department of Women’s and Children’s Health, Karolinska Institutet, Stockholm, Sweden

    Markus Kruusmägi

  6. A.N.Bach Institute of Biochemistry Russian Academy of Sciences, Moscow, Russia

    Alexandr V. Lopachev

  7. Department of Medicinal and Biological Chemistry, College of Pharmacy, University of Toledo, Toledo, USA

    Zahoor A. Shah

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  1. Evgeny E. Akkuratov
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  2. Olga M. Lopacheva
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  6. Alexander A. Boldyrev
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Akkuratov, E.E., Lopacheva, O.M., Kruusmägi, M. et al. Functional Interaction Between Na/K-ATPase and NMDA Receptor in Cerebellar Neurons. Mol Neurobiol 52, 1726–1734 (2015). https://doi.org/10.1007/s12035-014-8975-3

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