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Disruption of the clock components CLOCK and BMAL1 leads to hypoinsulinaemia and diabetes

Abstract

The molecular clock maintains energy constancy by producing circadian oscillations of rate-limiting enzymes involved in tissue metabolism across the day and night1,2,3. During periods of feeding, pancreatic islets secrete insulin to maintain glucose homeostasis, and although rhythmic control of insulin release is recognized to be dysregulated in humans with diabetes4, it is not known how the circadian clock may affect this process. Here we show that pancreatic islets possess self-sustained circadian gene and protein oscillations of the transcription factors CLOCK and BMAL1. The phase of oscillation of islet genes involved in growth, glucose metabolism and insulin signalling is delayed in circadian mutant mice, and both Clock5,6 and Bmal17 (also called Arntl) mutants show impaired glucose tolerance, reduced insulin secretion and defects in size and proliferation of pancreatic islets that worsen with age. Clock disruption leads to transcriptome-wide alterations in the expression of islet genes involved in growth, survival and synaptic vesicle assembly. Notably, conditional ablation of the pancreatic clock causes diabetes mellitus due to defective β-cell function at the very latest stage of stimulus–secretion coupling. These results demonstrate a role for the β-cell clock in coordinating insulin secretion with the sleep–wake cycle, and reveal that ablation of the pancreatic clock can trigger the onset of diabetes mellitus.

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Figure 1: Cell autonomous oscillator in pancreas.
Figure 2: Diabetic phenotypes in 8-month-old circadian mutant mice.
Figure 3: Reduced islet size, proliferation and insulin release in 8–10-month-old circadian mutant mice.
Figure 4: Impaired glucose tolerance and islet insulin secretion in 2–4-month-old pancreas-specific circadian mutant mice.

References

  1. Green, C. B., Takahashi, J. S. & Bass, J. The meter of metabolism. Cell 134, 728–742 (2008)

    CAS  Article  Google Scholar 

  2. Panda, S. et al. Coordinated transcription of key pathways in the mouse by the circadian clock. Cell 109, 307–320 (2002)

    CAS  Article  Google Scholar 

  3. Rutter, J., Reick, M. & McKnight, S. L. Metabolism and the control of circadian rhythms. Annu. Rev. Biochem. 71, 307–331 (2002)

    CAS  Article  Google Scholar 

  4. Polonsky, K. S. et al. Abnormal patterns of insulin secretion in non-insulin-dependent diabetes mellitus. N. Engl. J. Med. 318, 1231–1239 (1988)

    CAS  Article  Google Scholar 

  5. King, D. P. et al. Positional cloning of the mouse circadian clock gene. Cell 89, 641–653 (1997)

    CAS  Article  Google Scholar 

  6. Turek, F. W. et al. Obesity and metabolic syndrome in circadian Clock mutant mice. Science 308, 1043–1045 (2005)

    ADS  CAS  Article  Google Scholar 

  7. Bunger, M. K. et al. Mop3 is an essential component of the master circadian pacemaker in mammals. Cell 103, 1009–1017 (2000)

    CAS  Article  Google Scholar 

  8. Lowrey, P. L. & Takahashi, J. S. Mammalian circadian biology: elucidating genome-wide levels of temporal organization. Annu. Rev. Genomics Hum. Genet. 5, 407–441 (2004)

    CAS  Article  Google Scholar 

  9. Kornmann, B. et al. System-driven and oscillator-dependent circadian transcription in mice with a conditionally active liver clock. PLoS Biol. 5, e34 (2007)

    Article  Google Scholar 

  10. McCarthy, J. J. et al. Identification of the circadian transcriptome in adult mouse skeletal muscle. Physiol. Genomics 31, 86–95 (2007)

    CAS  Article  Google Scholar 

  11. Storch, K. F. et al. Extensive and divergent circadian gene expression in liver and heart. Nature 417, 78–83 (2002)

    ADS  CAS  Article  Google Scholar 

  12. Yang, X. et al. Nuclear receptor expression links the circadian clock to metabolism. Cell 126, 801–810 (2006)

    CAS  Article  Google Scholar 

  13. Balsalobre, A., Damiola, F. & Schibler, U. A serum shock induces circadian gene expression in mammalian tissue culture cells. Cell 93, 929–937 (1998)

    CAS  Article  Google Scholar 

  14. Yoo, S. H. et al. PERIOD2::LUCIFERASE real-time reporting of circadian dynamics reveals persistent circadian oscillations in mouse peripheral tissues. Proc. Natl Acad. Sci. USA 101, 5339–5346 (2004)

    ADS  CAS  Article  Google Scholar 

  15. Yamazaki, S. et al. Resetting central and peripheral circadian oscillators in transgenic rats. Science 288, 682–685 (2000)

    ADS  CAS  Article  Google Scholar 

  16. Drucker, D. J. The biology of incretin hormones. Cell Metab. 3, 153–165 (2006)

    CAS  Article  Google Scholar 

  17. Gao, N. et al. Foxa2 controls vesicle docking and insulin secretion in mature β cells. Cell Metab. 6, 267–279 (2007)

    CAS  Article  Google Scholar 

  18. Rulifson, I. C. et al. Wnt signaling regulates pancreatic β cell proliferation. Proc. Natl Acad. Sci. USA 104, 6247–6252 (2007)

    ADS  CAS  Article  Google Scholar 

  19. Shih, D. Q. et al. Loss of HNF-1α function in mice leads to abnormal expression of genes involved in pancreatic islet development and metabolism. Diabetes 50, 2472–2480 (2001)

    CAS  Article  Google Scholar 

  20. Stoffers, D. A. et al. Insulinotropic glucagon-like peptide 1 agonists stimulate expression of homeodomain protein IDX-1 and increase islet size in mouse pancreas. Diabetes 49, 741–748 (2000)

    CAS  Article  Google Scholar 

  21. Withers, D. J. et al. Disruption of IRS-2 causes type 2 diabetes in mice. Nature 391, 900–904 (1998)

    ADS  CAS  Article  Google Scholar 

  22. Gu, G., Dubauskaite, J. & Melton, D. A. Direct evidence for the pancreatic lineage: NGN3+ cells are islet progenitors and are distinct from duct progenitors. Development 129, 2447–2457 (2002)

    CAS  PubMed  Google Scholar 

  23. Westgate, E. J. et al. Genetic components of the circadian clock regulate thrombogenesis in vivo . Circulation 117, 2087–2095 (2008)

    Article  Google Scholar 

  24. Lamia, K. A., Storch, K. F. & Weitz, C. J. Physiological significance of a peripheral tissue circadian clock. Proc. Natl Acad. Sci. USA 105, 15172–15177 (2008)

    ADS  CAS  Article  Google Scholar 

  25. Rudic, R. D. et al. BMAL1 and CLOCK, two essential components of the circadian clock, are involved in glucose homeostasis. PLoS Biol. 2, e377 (2004)

    Article  Google Scholar 

  26. Duez, H. et al. Regulation of bile acid synthesis by the nuclear receptor Rev-erbα. Gastroenterology 135, 689–698 (2008)

    CAS  Article  Google Scholar 

  27. Noshiro, M. et al. Multiple mechanisms regulate circadian expression of the gene for cholesterol 7α-hydroxylase (Cyp7a), a key enzyme in hepatic bile acid biosynthesis. J. Biol. Rhythms 22, 299–311 (2007)

    CAS  Article  Google Scholar 

  28. Kudo, T., Kawashima, M., Tamagawa, T. & Shibata, S. Clock mutation facilitates accumulation of cholesterol in the liver of mice fed a cholesterol and/or cholic acid diet. Am. J. Physiol. Endocrinol. Metab. 294, E120–E130 (2008)

    CAS  Article  Google Scholar 

  29. Fu, L. et al. The molecular clock mediates leptin-regulated bone formation. Cell 122, 803–815 (2005)

    CAS  Article  Google Scholar 

  30. Matsuo, T. et al. Control mechanism of the circadian clock for timing of cell division in vivo . Science 302, 255–259 (2003)

    ADS  CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank F. Turek, R. Allada and G. Bell for discussions and comments on the manuscript. We thank A. Kohsaka, E. Chen, J. Doering and C. Radosevich for their technical support, as well as the Biological Imaging Facility at Northwestern University and the Islet Biology Core of the University of Chicago DRTC. We thank D. Melton for the PdxCre mice. Work was supported by grants from the National Institute of Diabetes and Digestive and Kidney Diseases to K.M.R. and L.H.P.; the National Institutes of Health, Chicago Biomedical Consortium Searle Funds, and Juvenile Diabetes Research Foundation to J.B.; grant R37-ES-005703 from the National Institutes of Health to C.A.B.; and the National Institute of Mental Health to J.S.T.

Author information

Authors and Affiliations

Authors

Contributions

B.M. performed and analysed most of the experiments in this study, with technical assistance from Y.K., G.I., S.M. and C.O. E.D.B. and C.H.K. conducted and analysed real-time bioluminescence imaging experiments in isolated pancreatic islets. H.S. conducted immunostaining experiments. M.H.V. performed statistical analysis. J.P.L. conducted and analysed Ca2+ influx experiments. S.D.C. and L.J. performed statistical and gene ontogeny analysis of microarray data. C.A.B. provided Bmal1flx/flx mice. J.S.T., L.H.P., X.W., K.M.R., B.M. and J.B. provided critical intellectual input in the preparation of the manuscript. K.M.R., B.M., J.S.T. and J.B. wrote the paper.

Corresponding author

Correspondence to Joseph Bass.

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Competing interests

[Competing Interests: J.S.T. is an Investigator in the Howard Hughes Medical Institute and a co-founder of ReSet Therapeutics Inc., and J.S.T. and J.B. are members of its scientific advisory board. J.B. is also an advisor and receives support from Amylin Pharmaceuticals.]

Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Descriptions 1-5, Supplementary Figures S1-S9 with legends, Supplementary Tables S1-S3 and References. (PDF 2403 kb)

Supplementary Movie 1

Cell autonomous oscillator in pancreas. (MOV 8408 kb)

Continuous videomicroscopy monitoring of islet bioluminescence over 72 hours. Islets from Per2Luc mice were isolated via collagenase digestion and imaged as described in Supplementary Methods. The 72-hour epoch is continuously replayed.

Supplementary Movie 2

Cell autonomous oscillator in pancreas. (MOV 8397 kb)

Islets were harvested from a separate group of Per2Luc mice and monitored under identical conditions for 72 hours. The 72-hour epoch is continuously replayed.

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Marcheva, B., Ramsey, K., Buhr, E. et al. Disruption of the clock components CLOCK and BMAL1 leads to hypoinsulinaemia and diabetes. Nature 466, 627–631 (2010). https://doi.org/10.1038/nature09253

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