Letter | Published:

Prefrontal cortex output circuits guide reward seeking through divergent cue encoding

Nature volume 543, pages 103107 (02 March 2017) | Download Citation


The prefrontal cortex is a critical neuroanatomical hub for controlling motivated behaviours across mammalian species1,2,3. In addition to intra-cortical connectivity, prefrontal projection neurons innervate subcortical structures that contribute to reward-seeking behaviours, such as the ventral striatum and midline thalamus4. While connectivity among these structures contributes to appetitive behaviours5,6,7,8,9,10,11,12,13, how projection-specific prefrontal neurons encode reward-relevant information to guide reward seeking is unknown. Here we use in vivo two-photon calcium imaging to monitor the activity of dorsomedial prefrontal neurons in mice during an appetitive Pavlovian conditioning task. At the population level, these neurons display diverse activity patterns during the presentation of reward-predictive cues. However, recordings from prefrontal neurons with resolved projection targets reveal that individual corticostriatal neurons show response tuning to reward-predictive cues, such that excitatory cue responses are amplified across learning. By contrast, corticothalamic neurons gradually develop new, primarily inhibitory responses to reward-predictive cues across learning. Furthermore, bidirectional optogenetic manipulation of these neurons reveals that stimulation of corticostriatal neurons promotes conditioned reward-seeking behaviour after learning, while activity in corticothalamic neurons suppresses both the acquisition and expression of conditioned reward seeking. These data show how prefrontal circuitry can dynamically control reward-seeking behaviour through the opposing activities of projection-specific cell populations.

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  1. 1.

    Increased spontaneous activity produced by frontal lobe lesion in cats. Am. J. Physiol. 126, 158–161 (1939)

  2. 2.

    , & Neuronal correlates of goal-based motor selection in the prefrontal cortex. Science 301, 229–232 (2003)

  3. 3.

    et al. A prefrontal cortex–brainstem neuronal projection that controls response to behavioural challenge. Nature 492, 428–432 (2012)

  4. 4.

    Differential projections of the infralimbic and prelimbic cortex in the rat. Synapse 51, 32–58 (2004)

  5. 5.

    et al. Synaptic and behavioral profile of multiple glutamatergic inputs to the nucleus accumbens. Neuron 76, 790–803 (2012)

  6. 6.

    et al. Bidirectional modulation of incubation of cocaine craving by silent synapse-based remodeling of prefrontal cortex to accumbens projections. Neuron 83, 1453–1467 (2014)

  7. 7.

    & The circuitry mediating cocaine-induced reinstatement of drug-seeking behavior. J. Neurosci. 21, 8655–8663 (2001)

  8. 8.

    , & Prefrontal glutamate release into the core of the nucleus accumbens mediates cocaine-induced reinstatement of drug-seeking behavior. J. Neurosci. 23, 3531–3537 (2003)

  9. 9.

    et al. Cocaine-induced synaptic alterations in thalamus to nucleus accumbens projection. Neuropsychopharmacology 41, 2399–2410 (2016)

  10. 10.

    et al. Contrasting forms of cocaine-evoked plasticity control components of relapse. Nature 509, 459–464 (2014)

  11. 11.

    , & Infralimbic prefrontal cortex is responsible for inhibiting cocaine seeking in extinguished rats. J. Neurosci. 28, 6046–6053 (2008)

  12. 12.

    et al. Optogenetic inhibition of cocaine seeking in rats. Addict. Biol. 18, 50–53 (2013)

  13. 13.

    et al. Wiring and molecular features of prefrontal ensembles representing distinct experiences. Cell 165, 1776–1788 (2016)

  14. 14.

    & Reward expectation, orientation of attention and locus coeruleus-medial frontal cortex interplay during learning. Eur. J. Neurosci. 20, 791–802 (2004)

  15. 15.

    , , , & Prefrontal parvalbumin neurons in control of attention. Cell 164, 208–218 (2016)

  16. 16.

    et al. Influences of rewarding and aversive outcomes on activity in macaque lateral prefrontal cortex. Neuron 51, 861–870 (2006)

  17. 17.

    & Prefrontal neurons encode context-based response execution and inhibition in reward seeking and extinction. Proc. Natl Acad. Sci. USA 112, 9472–9477 (2015)

  18. 18.

    et al. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature 499, 295–300 (2013)

  19. 19.

    et al. Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo. Proc. Natl Acad. Sci. USA 101, 18206–18211 (2004)

  20. 20.

    , , & Topographical organization of the efferent projections of the medial prefrontal cortex in the rat: an anterograde tract-tracing study with Phaseolus vulgaris leucoagglutinin. J. Comp. Neurol. 290, 213–242 (1989)

  21. 21.

    et al. Role of projections from ventral medial prefrontal cortex to nucleus accumbens shell in context-induced reinstatement of heroin seeking. J. Neurosci. 32, 4982–4991 (2012)

  22. 22.

    , , , & Prelimbic to accumbens core pathway is recruited in a dopamine-dependent manner to drive cued reinstatement of cocaine seeking. J. Neurosci. 36, 8700–8711 (2016)

  23. 23.

    , & Enhanced c-Fos expression in superior colliculus, paraventricular thalamus and septum during learning of cue-reward association. Neuroscience 168, 706–714 (2010)

  24. 24.

    & A potential role for the paraventricular nucleus of the thalamus in mediating individual variation in Pavlovian conditioned responses. Front. Behav. Neurosci. 8, 79 (2014)

  25. 25.

    , & A temporal shift in the circuits mediating retrieval of fear memory. Nature 519, 460–463 (2015)

  26. 26.

    et al. The paraventricular thalamus controls a central amygdala fear circuit. Nature 519, 455–459 (2015)

  27. 27.

    & Cortical connectivity and sensory coding. Nature 503, 51–58 (2013)

  28. 28.

    & Limited collateralization of neurons in the rat prefrontal cortex that project to the nucleus accumbens. Neuroscience 97, 635–642 (2000)

  29. 29.

    et al. Visualization of cortical, subcortical and deep brain neural circuit dynamics during naturalistic mammalian behavior with head-mounted microscopes and chronically implanted lenses. Nat. Protocols 11, 566–597 (2016)

  30. 30.

    et al. Construction of implantable optical fibers for long-term optogenetic manipulation of neural circuits. Nat. Protocols 7, 12–23 (2011)

  31. 31.

    , , & Preferential transduction of neurons by canine adenovirus vectors and their efficient retrograde transport in vivo. FASEB J. 15, 2283–2285 (2001)

  32. 32.

    et al. Cre recombinase-mediated restoration of nigrostriatal dopamine in dopamine-deficient mice reverses hypophagia and bradykinesia. Proc. Natl Acad. Sci. USA 103, 8858–8863 (2006)

  33. 33.

    , , & SIMA: Python software for analysis of dynamic fluorescence imaging data. Front. Neuroinform. 8, 80 (2014)

  34. 34.

    A study of cross-validation and bootstrap for accuracy estimation and model selection. IJCAI (U. S.) 95, 1137–1143 (1995)

  35. 35.

    et al. Excitatory transmission from the amygdala to nucleus accumbens facilitates reward seeking. Nature 475, 377–380 (2011)

  36. 36.

    , & Multiple neuroanatomical tract-tracing using fluorescent Alexa Fluor conjugates of cholera toxin subunit B in rats. Nat. Protocols 4, 1157–1166 (2009)

  37. 37.

    Transneuronal circuit tracing with neurotropic viruses. Curr. Opin. Neurobiol. 18, 617–623 (2008)

  38. 38.

    & Anatomical analysis of afferent projections to the medial prefrontal cortex in the rat. Brain Struct. Funct. 212, 149–179 (2007)

  39. 39.

    , & Neurobiological dissociation of retrieval and reconsolidation of cocaine-associated memory. J. Neurosci. 33, 1271–1281 (2013)

  40. 40.

    & The Mouse Brain in Stereotactic Coordinates 3rd edn (Academic Press, 2007)

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We thank S. Smith and J. Stirman for helpful discussions. This study was funded by grants from the National Institutes of Health (NIDA: F32-DA041184, J.M.O.; R01-DA032750, G.D.S.; R01-DA038168, G.D.S.; NICHD: T32-HD079124, S.L.R.; NIMH: T32-MH093315, J.A.M.), the Brain and Behavior Research Foundation (G.D.S.), the Children’s Tumor Foundation (016-01-006, J.E.R.), the Foundation of Hope (G.D.S.), the UNC Neuroscience Center (Helen Lyng White Fellowship, V.M.K.N.), the UNC Neuroscience Center Microscopy Core (P30 NS045892), and the UNC Department of Psychiatry (G.D.S.).

Author information

Author notes

    • James M. Otis
    •  & Vijay M. K. Namboodiri

    These authors contributed equally to this work.


  1. Department of Psychiatry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA

    • James M. Otis
    • , Vijay M. K. Namboodiri
    • , Ana M. Matan
    • , Elisa S. Voets
    • , Emily P. Mohorn
    • , Oksana Kosyk
    • , Jenna A. McHenry
    • , J. Elliott Robinson
    • , Shanna L. Resendez
    • , Mark A. Rossi
    •  & Garret D. Stuber
  2. Neuroscience Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA

    • Vijay M. K. Namboodiri
    •  & Garret D. Stuber
  3. Neuroscience Curriculum, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA

    • J. Elliott Robinson
    •  & Garret D. Stuber
  4. Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA

    • Garret D. Stuber


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V.M.K.N. wrote codes for analyses. V.M.K.N., A.M.M., E.S.V., E.P.M., O.K., J.A.M., J.E.R., S.L.R. and M.A.R. provided technical assistance for in vivo optogenetics, histology and immunohistochemistry. J.M.O. performed experiments and surgeries. J.M.O., V.M.K.N. and G.D.S. designed the experiments, analysed and interpreted the data, and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Garret D. Stuber.

Extended data

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  1. 1.

    Representative video revealing calcium dynamics of dorsal medial PFC GCaMP6s-expressing neurons

    Injection of AAV-CaMKII-GCaMP6s resulted in dynamic GCaMP6s fluorescence from hundreds of visible PFC neurons in vivo (see methods for details). Data acquisition occurred at 2.5Hz, and this representative video is shown at 10X normal speed (25 frames per second; 485 x 508 pixels; 373μmß x 390μm).

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