Cell Stress and Chaperones

, Volume 21, Issue 5, pp 883–894 | Cite as

Regulation of gene expression by NFAT transcription factors in hibernating ground squirrels is dependent on the cellular environment

  • Yichi Zhang
  • Kenneth B. StoreyEmail author
Original Paper


Calcineurin is a calmodulin-stimulated phosphatase that regulates the nuclear translocation of nuclear factor of activated T cell (NFAT) c1-4 through dephosphorylation. We believe that this mechanism plays various roles in the remodeling and maintenance of Ictidomys tridecemlineatus skeletal muscle. During hibernation, bouts of torpor and arousal take place, and squirrels do not lose muscle mass despite being inactive. Protein expression of Ca2+ signaling proteins were studied using immunoblotting. A DNA-protein interaction ELISA technique was created to test the binding of NFATs in the nucleus to DNA probes containing the NFAT response element under environmental conditions reflective of those during hibernation. Calcineurin protein levels increased by 3.08-fold during torpor (compared to euthermic control), whereas calpain1 levels also rose by 3.66-fold during torpor. Calmodulin levels were elevated upon entering torpor. NFATc4 binding to DNA showed a 1.4-fold increase during torpor, and we found that this binding was further enhanced when 600 nM of Ca2+ was supplemented. We also found that decreasing the temperature of ELISAs resulted in progressive decreases in the binding of NFATs c1, c3, and c4 to DNA. In summary, calmodulin and calpain1 appear to activate calcineurin and NFATc4 during torpor. NFAT binding to target promoters is affected by intranuclear [Ca2+] and environmental temperatures. Therefore, Ca2+ signaling and temperature changes play key roles in regulation of the NFAT-calcineurin pathway in skeletal muscle of hibernating 13-lined ground squirrels over the torpor-arousal cycle, and they may contribute to the avoidance of disuse-induced muscle atrophy that occurs naturally in these animals.


Nuclear factor of activated T cells Calcium signaling Hibernation Ictidomys tridecemlineatus Immunoblot DPI-ELISA 



We thank Dr. J.M. Hallenbeck and Dr. D.C. McMullen (NINDS, NIH, Bethesda) for providing the tissue samples for this study. 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.

Supplementary material

12192_2016_713_MOESM1_ESM.docx (13 kb)
ESM 1 (DOCX 12 kb)


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© Cell Stress Society International 2016

Authors and Affiliations

  1. 1.Institute of Biochemistry and Department of BiologyCarleton UniversityOttawaCanada

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