Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Zebrafish Clock rhythmic expression reveals independent peripheral circadian oscillators

Nature Neuroscience volume 1, pages 701–707 (1998)Cite this article

Abstract

The only vertebrate clock gene identified by mutagenesis is mouse Clock , which encodes a bHLH-PAS transcription factor. We have cloned Clock in zebrafish and show that, in contrast to its mouse homologue, it is expressed with a pronounced circadian rhythm in the brain and in two defined pacemaker structures, the eye and the pineal gland. Clock oscillation was also found in other tissues, including kidney and heart. In these tissues, expression of Clock continues to oscillate in vitro . This demonstrates that self-sustaining circadian oscillators exist in several vertebrate organs, as was previously reported for invertebrates.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Conservation of zebrafish, mouse and Drosophila CLOCK.
Figure 2: Expression of the zebrafish Clock gene oscillates in pacemaker structures.
Figure 3: Clock expression oscillates in the zebrafish brain.
Figure 4: Clock expression in different zebrafish organs demonstrates the existence of independent circadian oscillators.

Similar content being viewed by others

References

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

    Article  CAS  Google Scholar 

  2. Antoch, M. P. et al. Functional identification of the mouse circadian Clock gene by transgenic BAC rescue. Cell 89, 655– 667 (1997).

    Article  CAS  Google Scholar 

  3. Vitaterna, M. H. et al. Mutagenesis and mapping of a mouse gene, Clock, essential for circadian behavior. Science 264, 719 –725 (1994).

    Article  CAS  Google Scholar 

  4. Sun, Z. S. et al. RIGUI, a putative mammalian ortholog of the Drosophila period gene. Cell 90, 1003–1011 (1997).

    Article  CAS  Google Scholar 

  5. Sassone-Corsi, P. Molecular clocks: mastering time by gene regulation. Nature 392, 871–874 (1998).

    CAS  PubMed  Google Scholar 

  6. Dunlap, J. C. Genetics and molecular analysis of circadian rhythms. Annu. Rev. Genet. 30, 579–601 ( 1996).

    Article  CAS  Google Scholar 

  7. Takahashi, J. S. Molecular neurobiology and genetics of circadian rhythms in mammals. Annu. Rev. Neurosci. 18, 531–553 (1995).

    Article  CAS  Google Scholar 

  8. Allada, R., White, N. E., So, W. V., Hall, J. C. & Rosbash, M. A mutant Drosophila homolog of mammalian Clock disrupts circadian rhythms and transcription of period and timeless. Cell 93, 791–804 ( 1998).

    Article  CAS  Google Scholar 

  9. Darlington, T. K. et al. Closing the circadian loop: CLOCK-induced transcription of its own inhibitors per and tim. Science 280, 1599–1603 (1998).

    Article  CAS  Google Scholar 

  10. Bae, K., Lee, C., Sidote, D., Chuang, K.-Y. & Edery, I. Circadian regulation of a Drosophila homolog of the mammalian Clock gene: PER and TIM function as positive regulators. Mol. Cell. Biol. 18, 6142–6151 (1998).

    Article  CAS  Google Scholar 

  11. Hardin, P. E., Hall, J. C. & Rosbash, M. Feedback of the Drosophila period gene product on circadian cycling of its messenger RNA levels. Nature 343, 536–540 (1990).

    Article  CAS  Google Scholar 

  12. Hardin, P. E., Hall, J. C. & Rosbash, M. Circadian oscillations in period gene mRNA levels are transcriptionally regulated. Proc. Natl. Acad. Sci. USA 89, 11711–11715 (1992).

    Article  CAS  Google Scholar 

  13. Sehgal, A. et al. Rhythmic expression of timeless: a basis for promoting circadian cycles in period gene autoregulation. Science 270, 808–810 (1995).

    Article  CAS  Google Scholar 

  14. Zerr, D. M., Hall, J. C., Rosbash, M. & Siwicki, K. K. Circadian fluctuations of period protein immunoreactivity in the CNS and the visual system of Drosophila. J. Neurosci. 10, 2749– 2762 (1990).

    Article  CAS  Google Scholar 

  15. Rutila, J. E. et al. CYCLE is a second bHLH-PAS clock protein essential for circadian rhythmicity and transcription of Drosophila period and timeless. Cell 93, 805–814 ( 1998).

    Article  CAS  Google Scholar 

  16. Hogenesch, J. B., Gu, Y. Z., Jain, S. & Bradfield, C. A. The basic-helix-loop-helix-PAS orphan MOP3 forms transcriptionally active complexes with circadian and hypoxia factors. Proc. Natl. Acad. Sci. USA 95, 5474–5479 (1998).

    Article  CAS  Google Scholar 

  17. Gekakis, N. et al. Role of the CLOCK protein in the mammalian circadian mechanism. Science 280, 1564–1569 (1998).

    Article  CAS  Google Scholar 

  18. Plautz, J. D., Kaneko, M., Hall, J. C. & Kay, S. A. Independent photoreceptive circadian clocks throughout Drosophila. Science 278 , 1632–1635 (1997).

    Article  CAS  Google Scholar 

  19. Moore, R. Y. & Eichler, V. B. Loss of a circadian adrenal corticosterone rhythm following suprachiasmatic lesions in the rat. Brain Res. 42, 201–206 ( 1972).

    Article  CAS  Google Scholar 

  20. Stephan, F. K. & Zucker, I. Circadian rhythms in drinking behavior and locomotor activity of rats. Proc. Natl. Acad. Sci. USA 69, 1583–1586 (1972).

    Article  CAS  Google Scholar 

  21. Tosini, G. & Menaker, M. Circadian rhythms in cultured mammalian retina. Science 272, 419– 421 (1996).

    Article  CAS  Google Scholar 

  22. Tei, H. et al. Circadian oscillation of a mammalian homologue of the Drosophila period gene. Nature 389, 512– 516 (1997).

    Article  CAS  Google Scholar 

  23. Albrecht, U., Sun, Z. S., Eichele, G. & Lee, C. C. A differential response of two putative mammalian circadian regulators, mper1 and mper2, to light. Cell 91, 1055– 1064 (1997).

    Article  CAS  Google Scholar 

  24. Shearman, L. P., Zylka, M. J., Weaver, D. R., Kolakowski, L. F. Jr & Reppert, S. M. Two period homologs: circadian expression and photic regulation in the suprachiasmatic nuclei. Neuron 19, 1261– 1269 (1997).

    Article  CAS  Google Scholar 

  25. Zylka, M. J., Shearman, L. P., Weaver, D. R. & Reppert, S. M. Three period homologs in mammals: differential light responses in the suprachiasmatic circadian clock and oscillating transcripts outside of brain. Neuron 20, 1103–1110 ( 1998).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  27. Cahill, G. M. Circadian regulation of melatonin production in cultured zebrafish pineal and retina. Brain Res. 708, 177– 181 (1996).

    Article  CAS  Google Scholar 

  28. Rajendran, R. R. et al. Zebrafish interphotoreceptor retinoid-binding protein: differential circadian expression among cone subtypes. J. Exp. Biol. 199, 2775–2787 (1996).

    CAS  PubMed  Google Scholar 

  29. Gonzalez, G. A. & Montminy, M. R. Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. Cell 59, 675–680 (1989).

    Article  CAS  Google Scholar 

  30. Foulkes, N. S. & Sassone-Corsi, P. Transcription factors coupled to the cAMP-signalling pathway. Biochim. Biophys. Acta 1288, F101–121 ( 1996).

    CAS  PubMed  Google Scholar 

  31. Shigeyoshi, Y. et al. Light-induced resetting of a mammalian circadian clock is associated with rapid induction of the mPer1 transcript. Cell 91, 1043–1053 (1997).

    Article  CAS  Google Scholar 

  32. Reppert, S. M. & Weaver, D. R. Forward genetic approach strikes gold: cloning of a mammalian clock gene. Cell 89, 487–490 ( 1997).

    Article  CAS  Google Scholar 

  33. Reppert, S. M. & Sauman, I. Period and timeless tango: a dance of two clock genes. Neuron 15, 983–986 (1995).

    Article  CAS  Google Scholar 

  34. Fonjallaz, P., Ossipow, V., Wanner, G. & Schibler, U. The two PAR leucine zipper proteins, TEF and DBP, display similar circadian and tissue-specific expression, but have different target promoter preferences. Embo J. 15, 351–362 ( 1996).

    Article  CAS  Google Scholar 

  35. Servillo, G., Della Fazia, M. A. & Viola-Magni, M. Variation of tyrosine aminotransferase expression during the day in rats of different ages. Biochem. Biophys. Res. Comm. 175, 104–109 (1991).

    Article  CAS  Google Scholar 

  36. Reppert, S. M. & Weaver, D. R. Melatonin madness. Cell 83, 1059–1062 (1995).

    Article  CAS  Google Scholar 

  37. Miyamoto, Y. & Sancar, A. Vitamin B2-based blue-light photoreceptors in the retinohypothalamic tract as the photoactive pigments for setting the circadian clock in mammals. Proc. Natl. Acad. Sci. USA 95, 6097–6102 (1998).

    Article  CAS  Google Scholar 

  38. Klein, D. C., Moore, R. Y. & Reppert, S. M. (eds) Suprachiasmatic Nucleus—The Mind's Clock (Oxford Univ. Press, Oxford, 1991).

    Google Scholar 

  39. Chomczynski, P. & Sacchi, N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162, 156– 159 (1987).

    Article  CAS  Google Scholar 

  40. Foulkes, N. S., Borrelli, E. & Sassone-Corsi, P. CREM gene: use of alternative DNA-binding domains generates multiple antagonists of cAMP-induced transcription. Cell 64, 739–749 (1991).

    Article  CAS  Google Scholar 

  41. Mellström, B., Naranjo, J. R., Foulkes, N. S., Lafarga, M. & Sassone-Corsi, P. Transcriptional response to cAMP in brain: specific distribution and induction of CREM antagonists. Neuron 10, 655–665 ( 1993).

    Article  Google Scholar 

Download references

Acknowledgements

We thank Joseph S. Takahashi, Nicolas Cermakian, Dario De Cesare, Lucia Monaco, Jean-Marie Garnier, Pilar Garcia-Villalba and Patrick Blader for discussions, advice and gifts of materials. We also acknowledge the technical assistance of Estelle Heitz, as well as Dominique Biellman, Odile Nkundwa, Nadine Fisher, Serge Vicaire and Frank Ruffenach. D.W. was supported by an EEC TMR fellowship. Our studies are funded by grants from CNRS, INSERM, CHUR, Rhône-Poulenc Rorer (Bioavenir), Fondation pour la Recherche Médicale and Association pour la Recherche sur le Cancer (P. S.-C.).

Author information

Author notes

  1. The first two authors contributed equally to this work.

Authors and Affiliations

  1. Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS-INSERM-ULP, 1 rue Laurent Fries, Illkirch Cédex, 67404, C.U. de Strasbourg, France

    David Whitmore, Nicholas S. Foulkes, Uwe Strähle & Paolo Sassone-Corsi

Authors
  1. David Whitmore
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nicholas S. Foulkes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Uwe Strähle
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paolo Sassone-Corsi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paolo Sassone-Corsi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Whitmore, D., Foulkes, N., Strähle, U. et al. Zebrafish Clock rhythmic expression reveals independent peripheral circadian oscillators. Nat Neurosci 1, 701–707 (1998). https://doi.org/10.1038/3703

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/3703

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing