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.

  • Letter
  • Published:

A single pair of interneurons commands the Drosophila feeding motor program

Abstract

Many feeding behaviours are the result of stereotyped, organized sequences of motor patterns. These patterns have been the subject of neuroethological studies1,2, such as electrophysiological characterization of neurons governing prey capture in toads1,3. However, technical limitations have prevented detailed study of the functional role of these neurons, a common problem for vertebrate organisms. Complexities involved in studies of whole-animal behaviour can be resolved in Drosophila, in which remote activation of brain cells by genetic means4 enables us to examine the nervous system in freely moving animals to identify neurons that govern a specific behaviour, and then to repeatedly target and manipulate these neurons to characterize their function. Here we show neurons that generate the feeding motor program in Drosophila. We carried out an unbiased screen using remote neuronal activation and identified a critical pair of brain cells that induces the entire feeding sequence when activated. These ‘feeding neurons’ (here abbreviated to Fdg neurons for brevity) are also essential for normal feeding as their suppression or ablation eliminates sugar-induced feeding behaviour. Activation of a single Fdg neuron induces asymmetric feeding behaviour and ablation of a single Fdg neuron distorts the sugar-induced feeding behaviour to become asymmetric, indicating the direct role of these neurons in shaping motor-program execution. Furthermore, recording neuronal activity and calcium imaging simultaneously during feeding behaviour5 reveals that the Fdg neurons respond to food presentation, but only in starved flies. Our results demonstrate that Fdg neurons operate firmly within the sensorimotor watershed, downstream of sensory and metabolic cues and at the top of the feeding motor hierarchy, to execute the decision to feed.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1: Thermogenetic activation reproduced coordinated natural feeding behaviour.
Figure 2: Thermogenetically induced food ingestion through the pharyngeal pump.
Figure 3: Identification of the Fdg neuron.
Figure 4: Functional analyses of Fdg neuron.

Similar content being viewed by others

References

  1. Ewert, J. P. Neural correlates of key stimulus and releasing mechanism: a case study and two concepts. Trends Neurosci. 20, 332–339 (1997)

    Article  MathSciNet  CAS  Google Scholar 

  2. Dethier, V. G. Hungry Fly (Harvard University Press, 1976)

    Google Scholar 

  3. Matsushima, T., Satou, M. & Ueda, K. Medullary reticular neurons in the Japanese toad: morphologies and excitatory inputs from the optic tectum. J. Comp. Physiol. A 166, 7–22 (1989)

    Article  CAS  Google Scholar 

  4. Lima, S. Q. & Miesenbock, G. Remote control of behavior through genetically targeted photostimulation of neurons. Cell 121, 141–152 (2005)

    Article  CAS  Google Scholar 

  5. Yoshihara, M. Simultaneous recording of calcium signals from identified neurons and feeding behavior of Drosophila melanogaster. J. Vis. Exp. 62, e3625 (2012)

    Google Scholar 

  6. Yoshihara, M. & Ito, K. Improved Gal4 screening kit for large-scale generation of enhancer-trap strains. Drosoph. Inf. Serv. 83, 199–202 (2000)

    Google Scholar 

  7. O’Kane, C. J. & Gehring, W. J. Detection in situ of genomic regulatory elements in Drosophila. Proc. Natl Acad. Sci. USA 84, 9123–9127 (1987)

    Article  ADS  Google Scholar 

  8. Peabody, N. C. et al. Characterization of the decision network for wing expansion in Drosophila using targeted expression of the TRPM8 channel. J. Neurosci. 29, 3343–3353 (2009)

    Article  CAS  Google Scholar 

  9. Hamada, F. N. et al. An internal thermal sensor controlling temperature preference in Drosophila. Nature 454, 217–220 (2008)

    Article  ADS  CAS  Google Scholar 

  10. Singh, R. N. Neurobiology of the gustatory systems of Drosophila and some terrestrial insects. Microsc. Res. Tech. 39, 547–563 (1997)

    Article  CAS  Google Scholar 

  11. Tinbergen, N. The Study of Instinct (Clarendon Press, 1989)

    Google Scholar 

  12. Miller, A. in Biology of Drosophila (ed. Demerec, M. ) 420–534 (John Wiley & Sons, 1950)

    Google Scholar 

  13. Ferris, G. F. in Biology of Drosophila (ed. Demerec, M. ) 368–419 (John Wiley & Sons, 1950)

    Google Scholar 

  14. Gajewski, K. M. & Schulz, R. A. CF2 represses Actin 88F gene expression and maintains filament balance during indirect flight muscle development in Drosophila. PLoS ONE 5, e10713 (2010)

    Article  ADS  Google Scholar 

  15. Struhl, G. & Basler, K. Organizing activity of wingless protein in Drosophila. Cell 72, 527–540 (1993)

    Article  CAS  Google Scholar 

  16. Wang, Z., Singhvi, A., Kong, P. & Scott, K. Taste representations in the Drosophila brain. Cell 117, 981–991 (2004)

    Article  CAS  Google Scholar 

  17. Thorne, N., Chromey, C., Bray, S. & Amrein, H. Taste perception and coding in Drosophila. Curr. Biol. 14, 1065–1079 (2004)

    Article  CAS  Google Scholar 

  18. Miyazaki, T. & Ito, K. Neural architecture of the primary gustatory center of Drosophila melanogaster visualized with GAL4 and LexA enhancer-trap systems. J. Comp. Neurol. 518, 4147–4181 (2010)

    Article  Google Scholar 

  19. Hagiwara, S., Miyazaki, S., Moody, W. & Patlak, J. Blocking effects of barium and hydrogen ions on the potassium current during anomalous rectification in the starfish egg. J. Physiol. 279, 167–185 (1978)

    Article  CAS  Google Scholar 

  20. Tian, L. et al. Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators. Nature Methods 6, 875–881 (2009)

    Article  CAS  Google Scholar 

  21. Gordon, M. D. & Scott, K. Motor control in a Drosophila taste circuit. Neuron 61, 373–384 (2009)

    Article  CAS  Google Scholar 

  22. Manzo, A., Silies, M., Gohl, D. M. & Scott, K. Motor neurons controlling fluid ingestion in Drosophila. Proc. Natl Acad. Sci. USA 109, 6307–6312 (2012)

    Article  ADS  CAS  Google Scholar 

  23. Aponte, Y., Atasoy, D. & Sternson, S. M. AGRP neurons are sufficient to orchestrate feeding behavior rapidly and without training. Nature Neurosci. 14, 351–355 (2011)

    Article  CAS  Google Scholar 

  24. Wiersma, C. A. & Ikeda, K. Interneurons commanding swimmeret movements in the Crayfish, Procambarus clarki (Girard). Comp. Biochem. Physiol. 12, 509–525 (1964)

    Article  CAS  Google Scholar 

  25. Marder, E. & Bucher, D. Understanding circuit dynamics using the stomatogastric nervous system of lobsters and crabs. Annu. Rev. Physiol. 69, 291–316 (2007)

    Article  CAS  Google Scholar 

  26. Kimura, K., Hachiya, T., Koganezawa, M., Tazawa, T. & Yamamoto, D. Fruitless and doublesex coordinate to generate male-specific neurons that can initiate courtship. Neuron 59, 759–769 (2008)

    Article  CAS  Google Scholar 

  27. Yoshihara, M., Rheuben, M. B. & Kidokoro, Y. Transition from growth cone to functional motor nerve terminal in Drosophila embryos. J. Neurosci. 17, 8408–8426 (1997)

    Article  CAS  Google Scholar 

  28. Yoshihara, M. et al. Selective effects of neuronal-synaptobrevin mutations on transmitter release evoked by sustained versus transient Ca2+ increases and by cAMP. J. Neurosci. 19, 2432–2441 (1999)

    Article  CAS  Google Scholar 

  29. Yoshihara, M., Adolfsen, B., Galle, K. T. & Littleton, J. T. Retrograde signaling by Syt 4 induces presynaptic release and synapse-specific growth. Science 310, 858–863 (2005)

    Article  ADS  CAS  Google Scholar 

  30. 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 

  31. Shang, Y., Griffith, L. C. & Rosbash, M. Light-arousal and circadian photoreception circuits intersect at the large PDF cells of the Drosophila brain. Proc. Natl Acad. Sci. USA 105, 19587–19594 (2008)

    Article  ADS  CAS  Google Scholar 

  32. Yang, C. H. et al. Control of the postmating behavioral switch in Drosophila females by internal sensory neurons. Neuron 61, 519–526 (2009)

    Article  CAS  Google Scholar 

  33. Pauli, A. et al. Cell-type-specific TEV protease cleavage reveals cohesin functions in Drosophila neurons. Dev. Cell 14, 239–251 (2008)

    Article  CAS  Google Scholar 

  34. Kitamoto, T. Conditional disruption of synaptic transmission induces male-male courtship behavior in Drosophila. Proc. Natl Acad. Sci. USA 99, 13232–13237 (2002)

    Article  ADS  CAS  Google Scholar 

  35. Fouquet, W. et al. Maturation of active zone assembly by Drosophila Bruchpilot. J. Cell Biol. 186, 129–145 (2009)

    Article  CAS  Google Scholar 

  36. Leiss, F. et al. Characterization of dendritic spines in the Drosophila central nervous system. Dev. Neurobiol. 69, 221–234 (2009)

    Article  CAS  Google Scholar 

  37. Estes, P. S., Ho, G. L., Narayanan, R. & Ramaswami, M. Synaptic localization and restricted diffusion of a Drosophila neuronal synaptobrevin–green fluorescent protein chimera in vivo. J. Neurogenet. 13, 233–255 (2000)

    Article  CAS  Google Scholar 

  38. Fischler, W., Kong, P., Marella, S. & Scott, K. The detection of carbonation by the Drosophila gustatory system. Nature 448, 1054–1057 (2007)

    Article  ADS  CAS  Google Scholar 

  39. Baines, R. A., Uhler, J. P., Thompson, A., Sweeney, S. T. & Bate, M. Altered electrical properties in Drosophila neurons developing without synaptic transmission. J. Neurosci. 21, 1523–1531 (2001)

    Article  CAS  Google Scholar 

  40. McGuire, S. E., Le, P. T., Osborn, A. J., Matsumoto, K. & Davis, R. L. Spatiotemporal rescue of memory dysfunction in Drosophila. Science 302, 1765–1768 (2003)

    Article  ADS  CAS  Google Scholar 

  41. Awasaki, T., Lai, S. L., Ito, K. & Lee, T. Organization and postembryonic development of glial cells in the adult central brain of Drosophila. J. Neurosci. 28, 13742–13753 (2008)

    Article  CAS  Google Scholar 

  42. Hayashi, S. et al. GETDB, a database compiling expression patterns and molecular locations of a collection of Gal4 enhancer traps. Genesis 34, 58–61 (2002)

    Article  CAS  Google Scholar 

  43. Shiraiwa, T. & Carlson, J. R. Proboscis extension response (PER) assay in Drosophila. J. Vis. Exp. 3, 193 (2007)

    Google Scholar 

  44. Kubo, Y., Baldwin, T. J., Jan, Y. N. & Jan, L. Y. Primary structure and functional expression of a mouse inward rectifier potassium channel. Nature 362, 127–133 (1993)

    Article  ADS  CAS  Google Scholar 

  45. Marella, S. et al. Imaging taste responses in the fly brain reveals a functional map of taste category and behavior. Neuron 49, 285–295 (2006)

    Article  CAS  Google Scholar 

  46. Zar, J. Biostatistical Analysis 4th edn (Prentice Hall, 1999)

    Google Scholar 

Download references

Acknowledgements

We thank S. Waddell for discussions, fly stocks and reading of the manuscript; T. Lee for discussions and fly stocks; A. Sakurai for reading of the manuscript; S. Reppert for support; M. Alkema and T. Ip for discussions; members of the NP consortium for NP lines; T. Awasaki., C. Kao, V. Budnik, P. Garrity, M. Freeman, M. Rosbash, Y.-N. Jan, L. Luo, S. Sigrist, K. Scott, T. Tanimura, L. Looger, M. Ramaswami and K. Gajewski for fly stocks; K. Ikeda, T. Tanimura and H. Ishimoto for technical advices; A.Taylor and R. Seeham for technical help; and N. Yoshihara for material information. This work was supported by National Institute of Mental Health Grant MH85958, and the Worcester Foundation (to M.Y.), the National Institute of Mental Health Intramural Research Program (B.W.), the summer program of the Japan Society for the Promotion of Science/National Science Foundation (to T.F.F.), and a Japan Science and Technology Agency CREST grant (to K.I.).

Author information

Authors and Affiliations

Authors

Contributions

M.Y., S.I., T.F.F. and M.G. designed the research. T.F.F. screened GAL4 lines under the supervision of K.I and M.Y. T.F.F., S.I and M.Y. performed behavioural analyses. S.I. performed analyses of ingestion and pump movement while M.Y. visualized the pump movement. M.G., S.I., K.I. and M.Y. performed neuroanatomy. S.I. and M.Y. did the fly genetics. M.Y. performed calcium imaging. S.I. and M.Y. performed experiments of laser activation and laser ablation with the technical assistance of M.G. B.W. contributed TRPM8 and essential advice. M.Y., M.G. and T.F.F. wrote the paper with assistance from S.I., B.W. and K.I.

Corresponding author

Correspondence to Motojiro Yoshihara.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-17, Supplementary Tables 1-2, Supplementary Note 1, Supplementary Methods and additional references. (PDF 10764 kb)

Natural Drosophila feeding behaviour

WT flies were starved for 24 hours. See Figure 1 and text. (MOV 9114 kb)

Genetically-induced feeding behaviour

NP883>TrpA1 satiated flies were activated by restrictive temperature. All aspects of natural feeding behaviour were reproduced. (MOV 16186 kb)

Movement of pharyngeal pump and food ingestion by the pump

Movement of the pharyngeal pump was visualized by Myosin heavy chain (Mhc)-GFP and blue dye. Induced feeding of a NP883>TrpA1 fly at 29˚C was compared with natural feeding of a starved WT fly on 100 mM sucrose at 29˚C. At this high temperature, pump movement in natural feeding is two times faster (6-8 Hz) than that at 21˚C (3-4 Hz). Pattern of muscle contraction in the induced feeding at 29˚C was quite similar to that in the natural feeding at 29˚C. See Fig. 2, Supplementary Figure 7 and text. (MOV 26601 kb)

Feeding behaviour by activation of Fdg-neuron in flip-out flies

The first fly with TrpA1 expression in an Fdg-neuron at the fly’s left almost exclusively was activated by raising temperature (fly is same as in Figure 3d and Supplementary Figures 10c, d, e). All aspects of natural feeding behaviour were reproduced in a substantially coordinated manner (see text). See Supplementary Table 2 for information of all flies from the flip-out Gal80 screening in this video. See Figure 3, Supplementary Figure 10 and text. (MOV 14650 kb)

3D-structure of Fdg-neuron

See Figure 3, Supplementary Figure 12 and text. (MOV 9156 kb)

Suppression of feeding behaviour by Kir

Comparison of PER and feeding on food in free running condition between Kir and WT flies, See Supplementary Figure 14. In free running condition, a starved Kir fly rarely extends their proboscis to the food, and when it does, it does not continue. (MOV 4279 kb)

PER simultaneous with Ca2+ imaging at the cell body of Fdg-neuron

Illumination is provided by the confocal microscope laser used for GCaMP Ca2+ imaging. The proboscis of a NP883>GCaMP3.0 fly was immobilized by glue, and 400 mM sucrose solution on Washi wick was used for stimulation, with its head capsule open to expose the SEG for Ca2+ imaging. Simultaneous Ca2+ imaging is shown with the labella opening. See Figure 4 and text. (MOV 3094 kb)

Laser activation of Fdg-neuron

Laser-activation of an Fdg-neuron induced proboscis extension and pump movement in NP883>TrpA1; mCD8-GFP flies. Infrared illumination is detected by the CCD camera as a red-tint flash. See Figure 4 and text. (MOV 25944 kb)

Effect of laser-ablating Fdg-neurons on sucrose-induced proboscis extension

Two examples are shown. The first fly; Proboscis extension before (to the front) and after laser-ablation of the fly’s left Fdg-neuron (to the right) and after ablation of both Fdg-neurons (no response) in a NP883>mCD8-GFP fly. Ablation at the opposite side for the second fly. See Figure 4 and text. (MOV 27934 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Flood, T., Iguchi, S., Gorczyca, M. et al. A single pair of interneurons commands the Drosophila feeding motor program. Nature 499, 83–87 (2013). https://doi.org/10.1038/nature12208

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature12208

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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