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Coupled oscillators control morning and evening locomotor behaviour of Drosophila

Abstract

Daily rhythms of physiology and behaviour are precisely timed by an endogenous circadian clock1,2. These include separate bouts of morning and evening activity, characteristic of Drosophila melanogaster and many other taxa, including mammals3,4,5. Whereas multiple oscillators have long been proposed to orchestrate such complex behavioural programmes6, their nature and interplay have remained elusive. By using cell-specific ablation, we show that the timing of morning and evening activity in Drosophila derives from two distinct groups of circadian neurons: morning activity from the ventral lateral neurons that express the neuropeptide PDF, and evening activity from another group of cells, including the dorsal lateral neurons. Although the two oscillators can function autonomously, cell-specific rescue experiments with circadian clock mutants indicate that they are functionally coupled.

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Figure 1: Suppression of GAL4 activity with GAL80 in Drosophila circadian neurons, as examined with a UAS-ANFGFP reporter.
Figure 2: Genetic ablation of different groups of circadian neurons causes the loss of different locomotor activity components.
Figure 3: Targeted genetic ablation removes specific circadian neurons.
Figure 4: Functional disruption of the two oscillators.
Figure 5: Coupling of the two Drosophila oscillators CRY+PDF- and PDF+ underlies the coordination of the two daily activity peaks.

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References

  1. Dunlap, J. C. Molecular bases for circadian clocks. Cell 96, 271–290 (1999)

    Article  CAS  Google Scholar 

  2. Panda, S., Hogenesch, J. & Kay, S. Circadian rhythms from flies to human. Nature 417, 329–335 (2002)

    Article  ADS  CAS  Google Scholar 

  3. Wheeler, D. A., Hamblen-Coyle, M. J., Dushay, M. S. & Hall, J. C. Behavior in light-dark cycles of Drosophila mutants that are arrhythmic, blind, or both. J. Biol. Rhythms 8, 67–94 (1993)

    Article  CAS  Google Scholar 

  4. Helfrich-Forster, C. Differential control of morning and evening components in the activity rhythm of Drosophila melanogaster—sex specific differences suggest a different quality of activity. J. Biol. Rhythms 2, 135–154 (2000)

    Article  Google Scholar 

  5. Hall, J. Genetics and molecular biology of rhythms in Drosophila and other insects. Adv. Genet. 48, 1–280 (2003)

    Article  CAS  Google Scholar 

  6. Pittendrigh, C. S. & Daan, S. A functional analysis of circadian pacemakers in nocturnal rodents. V. Pacemaker structure: A clock for all seasons. J. Comp. Physiol. 106, 333–355 (1976)

    Article  Google Scholar 

  7. Helfrich-Forster, C. The locomotor activity rhythm of Drosophila melanogaster is controlled by a dual oscillator system. J. Insect Physiol. 47, 877–887 (2001)

    Article  CAS  Google Scholar 

  8. Kaneko, M. & Hall, J. C. Neuroanatomy of cells expressing clock genes in Drosophila: Transgenic manipulation of the period and timeless genes to mark the perikarya of circadian pacemaker neurons and their projections. J. Comp. Neurol. 422, 66–94 (2000)

    Article  CAS  Google Scholar 

  9. Helfrich-Forster, C. The Period clock gene is expressed in central nervous system neurons which also produce a neuropeptide that reveals the projections of circadian pacemaker cells within the brain of Drosophila melanogaster. Proc. Natl Acad. Sci. USA 92, 612–616 (1995)

    Article  ADS  CAS  Google Scholar 

  10. Renn, S. C. P., Park, J. H., Rosbash, M., Hall, J. C. & Taghert, P. H. A pdf neuropeptide gene mutation and ablation of PDF neurons each cause severe abnormalities of behavioral circadian rhythms in Drosophila. Cell 99, 791–802 (1999)

    Article  CAS  Google Scholar 

  11. Park, J. H. et al. Differential regulation of circadian pacemaker output by separate clock genes in Drosophila. Proc. Natl Acad. Sci. USA 97, 3608–3613 (2000)

    Article  ADS  CAS  Google Scholar 

  12. Peng, Y., Stoleru, D., Levine, J. D., Hall, J. C. & Rosbash, M. Drosophila free-running rhythms require intercellular communication. PLoS Biol. 1, E13 (2003)

    Article  Google Scholar 

  13. Emery, P. et al. Drosophila CRY is a deep brain circadian photoreceptor. Neuron 26, 493–504 (2000)

    Article  CAS  Google Scholar 

  14. Klarsfeld, A. et al. Novel features of cryptochrome-mediated photoreception in the brain circadian clock of Drosophila. J. Neurosci. 24, 1468–1477 (2004)

    Article  CAS  Google Scholar 

  15. Zhao, J. et al. Drosophila clock can generate ectopic circadian clocks. Cell 113, 755–766 (2003)

    Article  CAS  Google Scholar 

  16. Rao, S., Lang, C., Levitan, E. S. & Deitcher, D. L. Visualization of neuropeptide expression, transport, and exocytosis in Drosophila melanogaster. J. Neurobiol. 49, 159–172 (2001)

    Article  CAS  Google Scholar 

  17. Blanchardon, E. et al. Defining the role of Drosophila lateral neurons in the control of circadian rhythms in motor activity and eclosion by targeted genetic ablation and PERIOD protein overexpression. Eur. J. Neurosci. 13, 871–888 (2001)

    Article  CAS  Google Scholar 

  18. Veleri, S., Brandes, C., Helfrich-Forster, C., Hall, J. C. & Stanewsky, R. A self-sustaining, light-entrainable circadian oscillator in the Drosophila brain. Curr. Biol. 13, 1758–1767 (2003)

    Article  CAS  Google Scholar 

  19. Lee, T. & Luo, L. Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis. Neuron 22, 451–461 (1999)

    Article  CAS  Google Scholar 

  20. 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  ADS  CAS  Google Scholar 

  21. Yang, Z. & Sehgal, A. Role of molecular oscillations in generating behavioral rhythms in Drosophila. Neuron 29, 453–467 (2001)

    Article  CAS  Google Scholar 

  22. Lee, H. S., Billings, H. J. & Lehman, M. N. The suprachiasmatic nucleus: a clock of multiple components. J. Biol. Rhythms 18, 435–449 (2003)

    Article  CAS  Google Scholar 

  23. Jagota, A., de la Iglesia, H. O. & Schwartz, W. J. Morning and evening circadian oscillations in the suprachiasmatic nucleus in vitro. Nature Neurosci. 3, 372–376 (2000)

    Article  CAS  Google Scholar 

  24. de la Iglesia, H. O., Meyer, J., Carpino, A. Jr & Schwartz, W. J. Antiphase oscillation of the left and right suprachiasmatic nuclei. Science 290, 799–801 (2000)

    Article  ADS  CAS  Google Scholar 

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

    Article  ADS  CAS  Google Scholar 

  26. Yoshii, T. et al. Drosophila cry(b) mutation reveals two circadian clocks that drive locomotor rhythm and have different responsiveness to light. J. Insect Physiol. 50, 479–488 (2004)

    Article  CAS  Google Scholar 

  27. Harmar, A. et al. The VPAC(2) receptor is essential for circadian function in the mouse suprachiasmatic nuclei. Cell 109, 497–508 (2002)

    Article  CAS  Google Scholar 

  28. Nitabach, M. N., Blau, J. & Holmes, T. C. Electrical silencing of Drosophila pacemaker neurons stops the free-running circadian clock. Cell 109, 485–495 (2002)

    Article  CAS  Google Scholar 

  29. Levine, J., Funes, P., Dowse, H. & Hall, J. Signal analysis of behavioral and molecular cycles. BMC Neurosci. 3, 1 (2002)

    Article  Google Scholar 

  30. Levine, J., Funes, P., Dowse, H. & Hall, J. Advanced analysis of a cryptochrome mutation's effects on the robustness and phase of molecular cycles in isolated peripheral tissues of Drosophila. BMC Neurosci. 3, 5 (2002)

    Article  Google Scholar 

  31. Grima, B., Chélot, E., Xia, R. & Rouyer, F. Morning and evening activity peaks are controlled by different clock neurons of the Drosophila brain. Nature doi:10.1038/nature02935 (this issue)

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Acknowledgements

We thank A. Sehgal for providing the UAS-per2-4 flies and L. Luo for tubulinP-GAL80 plasmid, as well as J. Levine, R. Allada, L. Griffith, P. Emery and M.R. laboratory members for discussion and critical comments on the manuscript. We are grateful to J. Hall, P. Nawathean and M. McDonald for their support and advice. We also thank E. Dougherty for assistance in confocal microscopy, and H. Felton for administrative assistance. The work was supported in part by grants from the NIH to M.R.

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Correspondence to Michael Rosbash.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Table S1

Quantitative description of GFP-expressing clock neurons with different combinations of circadian drivers. (DOC 36 kb)

Supplementary Table S2

Rescue of Pdf-GAL4/UAS-hid behavioral phenotype by Pdf-GAL80. (DOC 35 kb)

Supplementary Table S3

Rescue of cry-GAL4/UAS-hid behavioral phenotype by cry-GAL80. (DOC 46 kb)

Supplementary Table S4

Characterization of circadian locomotor behavior with selective oscillator ablation. (DOC 28 kb)

Supplementary Table S5

Circadian locomotor behavior with disabled oscillators. (DOC 27 kb)

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Stoleru, D., Peng, Y., Agosto, J. et al. Coupled oscillators control morning and evening locomotor behaviour of Drosophila. Nature 431, 862–868 (2004). https://doi.org/10.1038/nature02926

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