Abstract

The complex behaviors underlying reward seeking and consumption are integral to organism survival. The hypothalamus and mesolimbic dopamine system are key mediators of these behaviors, yet regulation of appetitive and consummatory behaviors outside of these regions is poorly understood. The central nucleus of the amygdala (CeA) has been implicated in feeding and reward, but the neurons and circuit mechanisms that positively regulate these behaviors remain unclear. Here, we defined the neuronal mechanisms by which CeA neurons promote food consumption. Using in vivo activity manipulations and Ca2+ imaging in mice, we found that GABAergic serotonin receptor 2a (Htr2a)-expressing CeA neurons modulate food consumption, promote positive reinforcement and are active in vivo during eating. We demonstrated electrophysiologically, anatomically and behaviorally that intra-CeA and long-range circuit mechanisms underlie these behaviors. Finally, we showed that CeAHtr2a neurons receive inputs from feeding-relevant brain regions. Our results illustrate how defined CeA neural circuits positively regulate food consumption.

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Acknowledgements

We thank T. Caudullo, E. Fejzulahi and B. Hoisl for their help with management of the animal colony; R. Kasper for technical help with the in vivo optogenetics setup; T. Ruff and D. del Toro for help with tissue clearing; M. Fischer for help with histology; M. Schwarz (Max Planck Institute for Medical Research) for providing Cre-dependent AAV-mCherry virus; S. Arber (Friedrich Miescher Institute for Biomedical Research) for providing Cre-dependent AAV-Synaptophysin-myc virus; and A. Ghanem for initial help with rabies viruses. We thank P. Sah and N. Gogolla for critical reading of this manuscript. Modified rabies virus was provided by the GT3 Core Facility of the Salk Institute with funding from the NIH–NCI CCSG (P30 014195), an NINDS R24 Core Grant and funding from the NEI. This study was supported by the Max-Planck Society and the Deutsche Forschungsgemeinschaft (Synergy to R.K. and SPP1665 to K.K.C. and R.K.).

Author information

Author notes

    • Amelia M Douglass
    • , Hakan Kucukdereli
    •  & Marion Ponserre

    These authors contributed equally to this work.

Affiliations

  1. Molecules-Signaling-Development, Max Planck Institute of Neurobiology, Martinsried, Germany.

    • Amelia M Douglass
    • , Hakan Kucukdereli
    • , Marion Ponserre
    • , Cornelia Strobel
    • , Pilar L Alcala Morales
    •  & Rüdiger Klein
  2. Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.

    • Milica Markovic
    • , Jan Gründemann
    •  & Andreas Lüthi
  3. Max von Pettenkofer-Institute & Gene Center, Ludwig-Maximilians-University Munich, Munich, Germany.

    • Karl-Klaus Conzelmann
  4. Munich Cluster for Systems Neurology (SyNergy), Munich, Germany.

    • Rüdiger Klein

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Contributions

A.M.D., H.K., M.P., M.M., J.G. and C.S. designed and analyzed experiments. A.M.D. performed behavior experiments. H.K. performed electrophysiology and assisted with behavior experiments. M.P. performed rabies tracing. H.K. and M.P. performed other tracing experiments. C.S. performed paired recordings and hM3D ex vivo verification. A.M.D., H.K., M.P. and P.L.A.M. performed histology. A.M.D., H.K., M.M. and J.G. performed Ca2+ imaging experiments. K.-K.C. provided rabies virus and shared expertise in its use for monosynaptic tracing. R.K. and A.L. supervised experiments. A.M.D., H.K., M.P. and R.K. wrote the manuscript, and all other authors provided input.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Rüdiger Klein.

Integrated supplementary information

Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–10.

  2. 2.

    Life Sciences Reporting Summary

Videos

  1. 1.

    Photostimulation of CeAHtr2a::ChR2 neurons at 20Hz evokes feeding-related motor behaviors.

    These include, licking of the walls and floor and chewing and gnawing motor sequences.

  2. 2.

    Representative move of Ca2+ activity (post-processing movie) (1x speed) in CeAHtr2a::GCaMP6s neurons synchronized with the behavior video during the free-feeding experiment.

    Upon the onset of eating after an overnight fast, increased Ca2+ activity is observed in CeAHtr2a:: GCaMP6s neurons.

  3. 3.

    Representative move of Ca2+ activity (post-processing movie) (1x speed) in CeAHtr2a::GCaMP6s neurons synchronized with the behavior video during the FR1 experiment.

    Key behavior events are denoted by text. Ca2+ activity is observed in CeAHtr2a:: GCaMP6s neurons during eating but not during the nose-poke that triggered release of the food pellet.

  4. 4.

    CeAHtr2a::ChR2 mice with bilateral optic fibres over the CeA strongly nose-poke for intracranial delivery of a 20Hz 473nm light pulse train.

  5. 5.

    Three-dimensional visualization (dorsal and lateral) of CeAHtr2a axonal projections labeled by an AAV-Flex-mCherry virus injected into the central amygdala of an Htr2a-Cre brain.

    High (red) and low (blue) mCherry fluorescence intensity is seen distinctly in the BNST, LH, vlPAG and PBN.

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DOI

https://doi.org/10.1038/nn.4623

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