Abstract
Hibernation is a highly seasonal physiological adaptation that allows brown bears (Ursus arctos) to survive extended periods of low food availability. Similarly, daily or circadian rhythms conserve energy by coordinating body processes to optimally match the environmental light/dark cycle. Brown bears express circadian rhythms in vivo and their cells do in vitro throughout the year, suggesting that these rhythms may play important roles during periods of negative energy balance. Here, we use time-series analysis of RNA sequencing data and timed measurements of ATP production in adipose-derived fibroblasts from active and hibernation seasons under two temperature conditions to confirm that rhythmicity was present. Culture temperature matching that of hibernation body temperature (34 °C) resulted in a delay of daily peak ATP production in comparison with active season body temperatures (37 °C). The timing of peaks of mitochondrial gene transcription was altered as were the amplitudes of transcripts coding for enzymes of the electron transport chain. Additionally, we observed changes in mean expression and timing of key metabolic genes such as SIRT1 and AMPK which are linked to the circadian system and energy balance. The amplitudes of several circadian gene transcripts were also reduced. These results reveal a link between energy conservation and a functioning circadian system in hibernation.
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Data availability
All newly generated circadian RNAseq data are available on NCBI under BioProject (PRJNA986183). Code is available on GitHub at (https://github.com/Jansenht/Vincent_JCPB_ursus).
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Acknowledgements
This work was supported by the Bear Research and Conservation Endowment at Washington State University, National Science Foundation Office of Polar Programs Grant to JLK (award number: 2312253), and the National Science Foundation Office of Polar Programs Post-Doctoral Fellowship to BWP (award number: 2138649). We would like to thank Marina Savenkova for her technical support in the laboratory and Sascha Duttke for sequencing our RNA samples. We would also like to acknowledge and thank all the volunteers and staff at the Washington State University Bear Center, especially Brandon Evans-Hutzenbiler, Heather Keepers, Chelsea Davis, Tony Carnahan, Jessie McCleary, and Heather Havelock and the WSU Kamiak high-performance computing cluster that make this research possible. The authors are also thankful to the two anonymous reviewers for the helpful comments.
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HTJ and CTR obtained funding; EPV, JLK, and HTJ designed the study; HTJ, CTR, and EPV performed the blood and tissue sampling; EPV and HTJ performed the metabolic flux analyses, and transcription experiments; EPV, JLK, BWP, and HTJ analyzed the data; the first draft of the manuscript was written by EPV, and all authors read and approved the final manuscript.
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Vincent, E.P., Perry, B.W., Kelley, J.L. et al. Circadian gene transcription plays a role in cellular metabolism in hibernating brown bears, Ursus arctos. J Comp Physiol B 193, 699–713 (2023). https://doi.org/10.1007/s00360-023-01513-5
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DOI: https://doi.org/10.1007/s00360-023-01513-5