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Melanopsin-dependent photo-perturbation reveals desynchronization underlying the singularity of mammalian circadian clocks

Nature Cell Biology volume 9, pages 13271334 (2007) | Download Citation



Singularity behaviour in circadian clocks1,2 — the loss of robust circadian rhythms following exposure to a stimulus such as a pulse of bright light — is one of the fundamental but mysterious properties of clocks. To quantitatively perturb and accurately measure the dynamics of cellular clocks3,4, we synthetically produced photo-responsiveness within mammalian cells by exogenously introducing the photoreceptor melanopsin5,6,7,8 and continuously monitoring the effect of photo-perturbation on the state of cellular clocks. Here we report that a critical light pulse drives cellular clocks into singularity behaviour. Our theoretical analysis consistently predicts and subsequent single-cell level observation directly proves that desynchronization of individual cellular clocks underlies singularity behaviour. Our theoretical framework also explains why singularity behaviours have been experimentally observed in various organisms, and it suggests that desynchronization is a plausible mechanism for the observable singularity of circadian clocks. Importantly, these in vitro and in silico findings are further supported by in vivo observations that desynchronization underlies the multicell-level amplitude decrease in the rat suprachiasmatic nucleus induced by critical light pulses.

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  1. 1.

    Unclocklike behaviour of biological clocks. Nature 253, 315–319 (1975).

  2. 2.

    The Geometry of Biological Time (Springer-Verlag, New York, 1980).

  3. 3.

    , & A serum shock induces circadian gene expression in mammalian tissue culture cells. Cell 93, 929–937 (1998).

  4. 4.

    & Involvement of the MAP kinase cascade in resetting of the mammalian circadian clock. Genes Dev. 14, 645–649 (2000).

  5. 5.

    , , , & Addition of human melanopsin renders mammalian cells photoresponsive. Nature 433, 741–745 (2005).

  6. 6.

    et al. Induction of photosensitivity by heterologous expression of melanopsin. Nature 433, 745–749 (2005).

  7. 7.

    et al. Illumination of the melanopsin signaling pathway. Science 307, 600–604 (2005).

  8. 8.

    & Melanopsin: another way of signaling light. Neuron 49, 331–339 (2006).

  9. 9.

    et al. A transcription factor response element for gene expression during circadian night. Nature 418, 534–539 (2002).

  10. 10.

    et al. System-level identification of transcriptional circuits underlying mammalian circadian clocks. Nature Genet. 37, 187–192 (2005).

  11. 11.

    et al. Feedback repression is required for mammalian circadian clock function. Nature Genet. 38, 312–319 (2006).

  12. 12.

    , , , & in Chronobiology: Biological Timekeeping (eds Dunlap, J. C., Loros, J. J. & DeCoursey, P. J.) 67–106 (Sinauer, Sunderland, Massachusetts, 2004).

  13. 13.

    Forty years of PRCs–what have we learned? Chronobiol. Int. 16, 711–743 (1999).

  14. 14.

    & Light pulses induce 'singular' behavior and shorten the period of the circadian phototaxis rhythm in the CW15 strain of Chlamydomonas. J. Biol. Rhythms 7, 313–327 (1992).

  15. 15.

    , , , & Bioluminescence imaging of individual fibroblasts reveals persistent, independently phased circadian rhythms of clock gene expression. Curr. Biol. 14, 2289–2295 (2004).

  16. 16.

    et al. Circadian gene expression in individual fibroblasts: cell-autonomous and self-sustained oscillators pass time to daughter cells. Cell 119, 693–705 (2004).

  17. 17.

    , & Light-induced suppression of endogenous circadian amplitude in humans. Nature 350, 59–62 (1991).

  18. 18.

    & Light-induced uncoupling of multioscillatory circadian system in a diurnal rodent, Asian chipmunk. Am. J. Physiol. 276, R1390–R1396 (1999).

  19. 19.

    et al. Synchronization and maintenance of timekeeping in suprachiasmatic circadian clock cells by neuropeptidergic signaling. Curr. Biol. 16, 599–605 (2006).

  20. 20.

    et al. Synchronization of cellular clocks in the suprachiasmatic nucleus. Science 302, 1408–1412 (2003).

  21. 21.

    , & Limit cycle models for circadian rhythms based on transcriptional regulation in Drosophila and Neurospora. J. Biol. Rhythms 14, 433–448 (1999).

  22. 22.

    & A molecular explanation for the long-term suppression of circadian rhythms by a single light pulse. Am. J. Physiol. Regul. Integr. Comp. Physiol. 280, R1206–R1212 (2001).

  23. 23.

    , & Molecular mechanism of suppression of circadian rhythms by a critical stimulus. EMBO J. (2006).

  24. 24.

    , , & Per1 and Per2 gene expression in the rat suprachiasmatic nucleus: circadian profile and the compartment-specific response to light. Neuroscience 94, 141–150 (1999).

  25. 25.

    , & Development of vasoactive intestinal peptide mRNA rhythm in the rat suprachiasmatic nucleus. J. Neurosci. 17, 3920–3931 (1997).

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This research was supported by intramural Grant-in-Aid from the Center for Developmental Biology (CDB), Director's Fund from CDB (H.R.U.), KAKENHI (Grant-in-Aid for Scientific Research) on Priority Areas 'Systems Genomics' from the Ministry of Education, Culture, Sports, Science and Technology of Japan (H.R.U. and H.U.), Grant-in-Aid for Scientific Research on 'Development of Basic Technology to Control Biological Systems Using Chemical Compounds' from the New Energy and Industrial Technology Development Organization (NEDO) of Japan (H.R.U.) and Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists 17-4169 (T.J.K). We thank for Wataru Kishimoto for technical supports on high-throughput monitoring system, Rikuhiro G. Yamada for support for data analysis, Maki Ukai-Tadenuma for technical support, Yoichi Minami for discussion for in vivo experiments, and Douglas Sipp Michael Royle, Danny Forger for critical comments on the manuscript. We also thank John J. Tyson for his suggestion regarding Winfree's original idea of desynchronization.

Author information

Author notes

    • Hideki Ukai
    •  & Tetsuya J. Kobayashi

    These authors contributed equally to this work.


  1. Laboratory for Systems Biology, Center for Developmental Biology, RIKEN, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.

    • Hideki Ukai
    • , Tetsuya J. Kobayashi
    • , Koh-hei Masumoto
    •  & Hiroki R. Ueda
  2. Functional Genomics Unit, Center for Developmental Biology, RIKEN, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.

    • Hiroki R. Ueda
  3. Department of Bioscience, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.

  4. Research Fellow of Japan Society for the Promotion of Science, Kinki University School of Medicine, 377-2 Ohno-Hagashi, Osaka-Sayama, Osaka 589-8511, Japan.

    • Tetsuya J. Kobayashi
  5. Department of Anatomy and Neurobiology, Kinki University School of Medicine, 377-2 Ohno-Hagashi, Osaka-Sayama, Osaka 589-8511, Japan.

    • Mamoru Nagano
    • , Mitsugu Sujino
    •  & Yasufumi Shigeyoshi
  6. Division of Biological Science, Graduate School of Science, Nagoya University and CREST & SORST, Japan Science and Technology Corporation, Furo-cho 1, Chikusa, Nagoya 464-8602, Japan.

    • Takao Kondo
  7. COE Unit of Circadian Systems, Division of Molecular Genetics, Department of Biological Sciences, Nagoya University Graduate School of Science, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.

    • Kazuhiro Yagita
  8. Department of Cell Biology and Neuroscience, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.

    • Kazuhiro Yagita


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K.Y. and H.R.U. developed the concept of synthetic implementation of photo-responsiveness within NIH3T3 cells by using melanopsin. T.K. designed a high-throughput monitoring device. H.U. constructed all materials used in this work and performed high-throughput real-time luciferase reporter assays, single-cell real-time bioluminescence imaging and pharmacological assays. T.J.K. developed the theory, data-analysis methods, and the image-analysis software used in this work, analysed high-throughput, single-cell real-time bioluminescence, in situ hybridization, and locomotor activity data, and performed single-cell real-time bioluminescence imaging. M.N. and M.S. performed behaviour analysis of rats. M.N., K.M. and Y.S. performed in situ hybridization and analysed the data. H.U., T.J.K. and H.R.U wrote the manuscript. All authors discussed the results and commented on the manuscript text.

Corresponding author

Correspondence to Hiroki R. Ueda.

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