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Constant darkness is a circadian metabolic signal in mammals

Nature volume 439, pages 340–343 (2006)Cite this article

Abstract

Environmental light is the ‘zeitgeber’ (time-giver) of circadian behaviour1. Constant darkness is considered a ‘free-running’ circadian state. Mammals encounter constant darkness during hibernation2. Ablation of the master clock synchronizer, the suprachiasmatic nucleus, abolishes torpor, a hibernation-like state, implicating the circadian clock in this phenomenon2,3. Here we report a mechanism by which constant darkness regulates the gene expression of fat catabolic enzymes in mice. Genes for murine procolipase (mClps) and pancreatic lipase-related protein 2 (mPlrp2 ) are activated in a circadian manner in peripheral organs during 12 h dark:12 h dark (DD) but not light–dark (LD) cycles. This mechanism is deregulated in circadian-deficient mPer1-/-/mPer2m/m mice. We identified circadian-regulated 5′-AMP, which is elevated in the blood of DD mice, as a key mediator of this response. Synthetic 5′-AMP induced torpor and mClps expression in LD animals. Torpor induced by metabolic stress was associated with elevated 5′-AMP levels in DD mice. Levels of glucose and non-esterified fatty acid in the blood are reversed in DD and LD mice. Induction of mClps expression by 5′-AMP in LD mice was reciprocally linked to blood glucose levels. Our findings uncover a circadian metabolic rhythm in mammals.

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Figure 1: Northern blot analysis of mClps and mPlrp2 expression in mice livers.
Figure 2: Elevated concentration of a circadian-regulated circulatory molecule in DD mice.
Figure 3: 5′-AMP-induced mClps expression and torpor in LD mice.
Figure 4: Torpor and blood 5′-AMP levels in DD mice under metabolic stress.

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Acknowledgements

We thank J. Lever for helpful comments, and J. Volmer for affinity-purified 5′-nucleotidase and pSK-5′NT. This work was supported in part by an NIH grant and the UTHSC Dean's fund to C.C.L. M.R.B. is supported in part by NIH funding. Author Contributions J.Z. carried out the described metabolic experiments, characterized peak 2 as 5′-AMP, and demonstrated that 5′-AMP induces torpor and expression of mClps in peripheral organs that is blocked by dipyridamole. K.K. screened and identified mClps/mPlrp2 expression in liver of DD mice. M.R.B. contributed insight into adenosine chemistry. C.C.L. conceived and directed the work and recognized the differential temporal profiles of peak 2 in DD and LD mice.

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Author notes

  1. Jianfa Zhang and Krista Kaasik: *These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Texas Health Science Center, Texas, 77030, Houston, USA

    Jianfa Zhang, Michael R. Blackburn & Cheng Chi Lee

  2. Department of Biotechnology, Institute of Molecular and Cell Biology, Tartu University, 51010, Tartu, Estonia

    Krista Kaasik

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Corresponding author

Correspondence to Cheng Chi Lee.

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Supplementary information

Supplementary Figure 1.

Micro-array of differential gene expression in liver between LD and DD mice using a 6k Unigene mouse cDNA library. (PDF 1013 kb)

Supplementary Figure 2.

Northern blot analysis of mClps expression on consecutive days. (PDF 1216 kb)

Supplementary Figure 3.

HPLC peaks in LD and DD mice (PDF 538 kb)

Supplementary Figure 4.

The circadian regulated signal is 5′-adenosine monophosphate. (PDF 119 kb)

Supplementary Figure 5.

Northern analysis of mClps induction by adenine nucleotides. (PDF 784 kb)

Supplementary Figure 6.

Metabolic analysis of DD versus LD mice. (PDF 261 kb)

Supplementary Figure 7.

Northern blot analysis of ecto-5′nucleotidase mRNA expression in livers of LD and DD mice. (PDF 332 kb)

Supplementary Figure 8.

5′ -AMP induction of mClps expression in LD mice is reciprocally linked to blood glucose levels. (PDF 517 kb)

Supplementary Figure Legends

Text to accompany the above Supplementary Figures. (DOC 24 kb)

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Zhang, J., Kaasik, K., Blackburn, M. et al. Constant darkness is a circadian metabolic signal in mammals. Nature 439, 340–343 (2006). https://doi.org/10.1038/nature04368

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