Wood frogs (Rana sylvatica LeConte) are the major model for studies of natural freeze tolerance by ectothermic vertebrates. Multiple biochemical adaptations support winter freezing survival but, to date, the protective role of chaperone proteins has received little attention. The present study analyzes responses to freezing, anoxia or dehydration exposures and recovery from these stresses by chaperone proteins in six wood frog organs: Five heat shock proteins (Hsc70, Hsp110, Hsp60, Hsp40, and Hsp10) and two glucose-regulated proteins (Grp78 and Grp94) were assessed. Hsc70 was upregulated in liver, muscle, heart and kidney (1.5-2.0 fold) during freezing and levels of its partner proteins also rose (Hsp110 in three tissues and Hsp40 in four tissues), these responses aligning most closely with comparable responses to anoxia rather than to dehydration. The resident chaperones of the endoplasmic reticulum (Grp78 and Grp94) also rose during freezing in liver and muscle (1.4-1.8 fold) but were suppressed in heart and skin, patterns that generally differed from responses to anoxia or dehydration. Elevated GRPs in liver may support the production and secretion of novel freeze responsive proteins. Increased levels of mitochondrial Hsp60 and Hsp10 (1.5-2.2 fold) occurred in most tissues during freezing and generally mimicked responses to anoxia. Overall, this study indicates that increased levels of chaperone proteins resident in multiple subcellular compartments contribute to stabilizing the cellular proteome during whole body freezing of wood frogs. These responses are probably derived from pre-existing amphibian defenses for stabilizing the proteome under environmental low oxygen or dehydration stresses.

amphibian environmental stress, anoxia, dehydration, freeze tolerance, glucose-regulated proteins, heat shock proteins
Journal of experimental zoology. Part A, Ecological and integrative physiology
Department of Biology

Storey, J, & Storey, K. (2019). In defense of proteins: Chaperones respond to freezing, anoxia, or dehydration stress in tissues of freeze tolerant wood frogs. Journal of experimental zoology. Part A, Ecological and integrative physiology, 331(7), 392–402. doi:10.1002/jez.2306