A craniofacial-specific monosynaptic circuit enables heightened affective pain

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Humans often rank craniofacial pain as more severe than body pain. Evidence suggests that a stimulus of the same intensity induces stronger pain in the face than in the body. However, the underlying neural circuitry for the differential processing of facial versus bodily pain remains unknown. Interestingly, the lateral parabrachial nucleus (PBL), a critical node in the affective pain circuit, is activated more strongly by noxious stimulation of the face than of the hindpaw. Using a novel activity-dependent technology called CANE developed in our laboratory, we identified and selectively labeled noxious-stimulus-activated PBL neurons and performed comprehensive anatomical input–output mapping. Surprisingly, we uncovered a hitherto uncharacterized monosynaptic connection between cranial sensory neurons and the PBL-nociceptive neurons. Optogenetic activation of this monosynaptic craniofacial-to-PBL projection induced robust escape and avoidance behaviors and stress calls, whereas optogenetic silencing specifically reduced facial nociception. The monosynaptic circuit revealed here provides a neural substrate for heightened craniofacial affective pain.

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We thank J. Takatoh for helping with a method to quantify axon innervation densities, K. Tschida and T. Gibson for helping with vocalization quantification and analysis, and V. Prevosto for helping with statistics. We also thank T. Gibson, M. Fu, K. Tschida, T. Stanek, V. Prevosto, and R. R. Ji for providing input and support throughout the project, and S. Lisberger and R. Mooney for critical reading of this manuscript. E.R. is supported by a F31 DE025197-03 fellowship. Y.C. is supported by K12DE022793. W.L. is supported by DE018549. This work is supported by NIH Grant DP1MH103908 to F.W.

Author information


  1. Department of Neurobiology, Duke University Medical Center, Durham, NC, USA

    • Erica Rodriguez
    • , Katsuyasu Sakurai
    • , Jennie Xu
    • , Shengli Zhao
    • , Bao-Xia Han
    • , David Ryu
    •  & Fan Wang
  2. Department of Neurology, Duke University Medical Center, Durham, NC, USA

    • Yong Chen
    •  & Wolfgang Liedtke
  3. Department of Psychology and Neuroscience, Duke University, Durham, NC, USA

    • Koji Toda
    •  & Henry Yin


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F.W. and E.R. conceived the idea and designed the experiments. E.R. performed the majority of the experiments and data analysis. K.S. performed some independent CANE capture experiments, bilateral fiber implantations and the place escape/avoidance (PEA) behavioral experiments. K.T. analyzed PEA results (blind to genotype). J.X. performed immunohistochemistry, quantified axon projections, and quantified cells in Fos and trans-synaptic experiments (blind to experimental conditions). Y.C. performed all the face and hindpaw von Frey assays (blind to genotypes). D.R. quantified cells in a subset of colocalization experiments. S.Z. produced all the CANE-LV and CANE-RV viruses. B.-X.H. took care of mouse husbandry and genotyping. H.Y. and W.L. provided critical equipment and reagents. F.W. and E.R. wrote the manuscript with help from W.L.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Fan Wang.

Integrated supplementary information

Supplementary information

  1. Supplementary Text and Figures

    Supplementary Figures 1–10

  2. Life Sciences Reporting Summary

  3. Supplementary Video 1

    Optogenetic activation of TrpV1Cre::ChR2+ TG afferents in the PBL in a real-time place escape/avoidance test (related to Fig. 4). Photo activation of TrpV1Cre::ChR2+ TG axon terminals within the PBL elicits escaping from the stimulation chamber to the opposite chamber to stop the stimulation. After mouse escapes to the non-stimulated chamber, it moves less and spends more time in the chamber.

  4. Supplementary Video 2

    Photo illumination of TrpV1Cre::GFP+ TG afferents in the PBL in a real-time place escape/avoidance test (related to Fig. 4). Photo illumination of TrpV1Cre::GFP+ TG axon terminals within the PBL has no observable behavioral effects.

  5. Supplementary Video 3

    Optogenetic activation of TrpV1Cre::ChR2+ TG afferents in the PBL in a circular chamber to record vocalization (related to Fig. 4). Photo activation of TrpV1Cre::ChR2+ TG axon terminals within the PBL induces audible distress vocalization. Vocalization stops when laser light turns off.

  6. Supplementary Video 4

    Photo illumination of TrpV1Cre::GFP+ TG afferents in the PBL in a circular chamber to record vocalization (related to Fig. 4). Photo illumination of TrpV1Cre::GFP+ TG axon terminals within the PBL does not induce any vocalizations.