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The role of DNA methylation during anoxia tolerance in a freshwater turtle (Trachemys scripta elegans)

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Abstract

Oxygen deprivation is a lethal stress that only a few animals can tolerate for extended periods. This study focuses on analyzing the role of DNA methylation in aiding natural anoxia tolerance in a champion vertebrate anaerobe, the red-eared slider turtle (Trachemys scripta elegans). We examined the relative expression and total enzymatic activity of four DNA methyltransferases (DNMT1, DNMT2, DNMT3a and DNMT3b), two methyl-binding domain proteins (MBD1 and MBD2), and relative genomic levels of 5-methylcytosine under control, 5 h anoxic, and 20 h anoxic conditions in liver, heart, and white skeletal muscle (n = 4, p < 0.05). In liver, protein expression of DNMT1, DNMT2, MBD1, and MBD2 rose significantly by two- to fourfold after 5 h anoxic submergence compared to normoxic-control conditions. In heart, 5 h anoxia submergence resulted in a 1.4-fold increase in DNMT3a levels and a significant decrease in MBD1 and MBD2 levels to ~30 % of control values. In white muscle, DNMT3a and DNMT3b increased threefold and MBD1 levels increased by 50 % in response to 5 h anoxia. Total DNMT activity rose by 0.6–2.0-fold in liver and white muscle and likewise global 5mC levels significantly increased in liver and white muscle under 5 and 20 h anoxia. The results demonstrate an overall increase in DNA methylation, DNMT protein expression and enzymatic activity in response to 5 and 20 h anoxia in liver and white muscle indicating a potential downregulation of gene expression via this epigenetic mechanism during oxygen deprivation.

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Acknowledgements

Thanks to J.M. Storey for editorial review of the manuscript. This work was supported by a Discovery grant from the Natural Sciences and Engineering Research Council (NSERC) of Canada (Grant #: 6793). KBS holds the Canada Research Chair in Molecular Physiology and SW holds a postgraduate Queen Elizabeth II Graduate Scholarship in Science and Technology.

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Authors and Affiliations

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

    Sanoji Wijenayake

  2. Department of Biology, Department of Chemistry, Canada Research Chair in Molecular Physiology, Institute of Biochemistry, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada

    Kenneth B. Storey

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  1. Sanoji Wijenayake
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  2. Kenneth B. Storey
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Correspondence to Kenneth B. Storey.

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Communicated by G. Heldmaier.

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Wijenayake, S., Storey, K.B. The role of DNA methylation during anoxia tolerance in a freshwater turtle (Trachemys scripta elegans). J Comp Physiol B 186, 333–342 (2016). https://doi.org/10.1007/s00360-016-0960-x

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  • DOI: https://doi.org/10.1007/s00360-016-0960-x

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