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Regulation of nuclear factor of activated T cells (NFAT) and downstream myogenic proteins during dehydration in the African clawed frog

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Abstract

Xenopus laevis, otherwise known as the African clawed frog, undergoes natural dehydration of up to 30% of its total body water during the dry season in sub-Saharan Africa. To survive under these conditions, a variety of physiological and biochemical changes take place in X. laevis. We were interested in understanding the role that the calcineurin-NFAT pathway plays during dehydration stress response in the skeletal muscles of X. laevis. Immunoblotting was performed to characterize the protein levels of NFATc1-4, calcium signalling proteins, in addition to myogenic proteins (MyoD, MyoG, myomaker). In addition, DNA–protein interaction ELISAs were used to assess the binding of NFATs to their consensus binding sequence, and to identify the effect of urea on NFAT-binding. Our results showed that NFATc1 and c4 protein levels decreased during dehydration, and there were no changes in NFATc2, c3, and calcium signalling proteins. However, MyoG and myomaker both showed increases in protein levels during dehydration, thus indicating that the late myogenic program involving myoblast differentiation, but not satellite cell activation and myoblast proliferation, could be involved in preserving the skeletal muscle of X. laevis during dehydration. In addition, we observed that urea seems to reduce NFATc3-binding to DNA during control, but not during dehydration, possibly indicating that NFATc3 is protected from the denaturing effects of urea as it accumulates during dehydration. These findings expand upon our knowledge of adaptive responses to dehydration, and they identify specific protein targets that could be used to protect the skeletal muscle from damage during stress.

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Acknowledgements

This work was supported by a Discovery grant from the Natural Sciences and Engineering Research Council (NSERC) of Canada (#6793) to Kenneth B. Storey. Kenneth B. Storey holds the Canada Research Chair in Molecular Physiology; Yichi Zhang held a postgraduate Queen Elizabeth II Graduate Scholarship in Science and Technology.

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Authors and Affiliations

  1. Institute of Biochemistry and Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada

    Yichi Zhang, Simon G. English & Kenneth B. Storey

  2. Department of Molecular Biology, Hamon Center for Regenerative Science and Medicine, Sen. Paul D. Wellstone Muscular Dystrophy Cooperative Research Center, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX, 75390, USA

    Yichi Zhang

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Correspondence to Kenneth B. Storey.

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Animals were cared for in accordance with the guidelines of the Canadian Council on Animal Care and all experimental procedures had the prior approval of the Carleton University Animal Care Committee (protocol #13683).

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Zhang, Y., English, S.G. & Storey, K.B. Regulation of nuclear factor of activated T cells (NFAT) and downstream myogenic proteins during dehydration in the African clawed frog. Mol Biol Rep 45, 751–761 (2018). https://doi.org/10.1007/s11033-018-4214-8

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