Corticotropin-releasing factor (CRF) that is released from the paraventricular nucleus (PVN) of the hypothalamus is essential for mediating stress response by activating the hypothalamic–pituitary–adrenal axis. CRF-releasing PVN neurons receive inputs from multiple brain regions that convey stressful events, but their neuronal dynamics on the timescale of behavior remain unknown. Here, our recordings of PVN CRF neuronal activity in freely behaving mice revealed that CRF neurons are activated immediately by a range of aversive stimuli. By contrast, CRF neuronal activity starts to drop within a second of exposure to appetitive stimuli. Optogenetic activation or inhibition of PVN CRF neurons was sufficient to induce a conditioned place aversion or preference, respectively. Furthermore, conditioned place aversion or preference induced by natural stimuli was significantly decreased by manipulating PVN CRF neuronal activity. Together, these findings suggest that the rapid, biphasic responses of PVN CRF neurons encode the positive and negative valences of stimuli.

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  1. 1.

    Namburi, P., Al-Hasani, R., Calhoon, G. G., Bruchas, M. R. & Tye, K. M. Architectural representation of valence in the limbic system. Neuropsychopharmacology 41, 1697–1715 (2016).

  2. 2.

    Tye, K. M. Neural circuit motifs in valence processing. Neuron 100, 436–452 (2018).

  3. 3.

    Ungless, M. A., Magill, P. J. & Bolam, J. P. Uniform inhibition of dopamine neurons in the ventral tegmental area by aversive stimuli. Science 303, 2040–2042 (2004).

  4. 4.

    Tsai, H. C. et al. Phasic firing in dopaminergic neurons is sufficient for behavioral conditioning. Science 324, 1080–1084 (2009).

  5. 5.

    Danjo, T., Yoshimi, K., Funabiki, K., Yawata, S. & Nakanishi, S. Aversive behavior induced by optogenetic inactivation of ventral tegmental area dopamine neurons is mediated by dopamine D2 receptors in the nucleus accumbens. Proc. Natl Acad. Sci. USA 111, 6455–6460 (2014).

  6. 6.

    Bains, J. S., Wamsteeker Cusulin, J. I. & Inoue, W. Stress-related synaptic plasticity in the hypothalamus. Nat. Rev. Neurosci. 16, 377–388 (2015).

  7. 7.

    Ulrich-Lai, Y. M. & Herman, J. P. Neural regulation of endocrine and autonomic stress responses. Nat. Rev. Neurosci. 10, 397–409 (2009).

  8. 8.

    Gunaydin, L. A. et al. Natural neural projection dynamics underlying social behavior. Cell 157, 1535–1551 (2014).

  9. 9.

    Cui, G. et al. Concurrent activation of striatal direct and indirect pathways during action initiation. Nature 494, 238–242 (2013).

  10. 10.

    Wamsteeker Cusulin, J. I., Füzesi, T., Watts, A. G. & Bains, J. S. Characterization of corticotropin-releasing hormone neurons in the paraventricular nucleus of the hypothalamus of Crh-IRES-Cre mutant mice. PLoS One 8, e64943 (2013).

  11. 11.

    Yilmaz, M. & Meister, M. Rapid innate defensive responses of mice to looming visual stimuli. Curr. Biol. 23, 2011–2015 (2013).

  12. 12.

    Fendt, M., Endres, T., Lowry, C. A., Apfelbach, R. & McGregor, I. S. TMT-induced autonomic and behavioral changes and the neural basis of its processing. Neurosci. Biobehav. Rev. 29, 1145–1156 (2005).

  13. 13.

    Golden, S. A., Covington, H. E. III, Berton, O. & Russo, S. J. A standardized protocol for repeated social defeat stress in mice. Nat. Protoc. 6, 1183–1191 (2011).

  14. 14.

    Seip, K. M. & Morrell, J. I. Exposure to pups influences the strength of maternal motivation in virgin female rats. Physiol. Behav. 95, 599–608 (2008).

  15. 15.

    Boyden, E. S., Zhang, F., Bamberg, E., Nagel, G. & Deisseroth, K. Millisecond-timescale, genetically targeted optical control of neural activity. Nat. Neurosci. 8, 1263–1268 (2005).

  16. 16.

    Aravanis, A. M. et al. An optical neural interface: in vivo control of rodent motor cortex with integrated fiberoptic and optogenetic technology. J. Neural. Eng. 4, S143–S156 (2007).

  17. 17.

    Füzesi, T., Daviu, N., Wamsteeker Cusulin, J. I., Bonin, R. P. & Bains, J. S. Hypothalamic CRH neurons orchestrate complex behaviours after stress. Nat. Commun. 7, 11937 (2016).

  18. 18.

    Tan, K. R. et al. GABA neurons of the VTA drive conditioned place aversion. Neuron 73, 1173–1183 (2012).

  19. 19.

    Chow, B. Y. et al. High-performance genetically targetable optical neural silencing by light-driven proton pumps. Nature 463, 98–102 (2010).

  20. 20.

    Mattis, J. et al. Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nat. Methods 9, 159–172 (2011).

  21. 21.

    Yasoshima, Y., Scott, T. R. & Yamamoto, T. Differential activation of anterior and midline thalamic nuclei following retrieval of aversively motivated learning tasks. Neuroscience 146, 922–930 (2007).

  22. 22.

    Zhu, Y., Wienecke, C. F. R., Nachtrab, G. & Chen, X. A thalamic input to the nucleus accumbens mediates opiate dependence. Nature 530, 219–222 (2016).

  23. 23.

    Betley, J. N. et al. Neurons for hunger and thirst transmit a negative-valence teaching signal. Nature 521, 180–185 (2015).

  24. 24.

    Chen, Y., Lin, Y. C., Kuo, T. W. & Knight, Z. A. Sensory detection of food rapidly modulates arcuate feeding circuits. Cell 160, 829–841 (2015).

  25. 25.

    Mandelblat-Cerf, Y. et al. Arcuate hypothalamic AgRP and putative POMC neurons show opposite changes in spiking across multiple timescales. eLife 4, 4 (2015).

  26. 26.

    Zimmerman, C. A. et al. Thirst neurons anticipate the homeostatic consequences of eating and drinking. Nature 537, 680–684 (2016).

  27. 27.

    Spruijt, B. M., van Hooff, J. A. & Gispen, W. H. Ethology and neurobiology of grooming behavior. Physiol. Rev. 72, 825–852 (1992).

  28. 28.

    Dunn, A. J. & Swiergiel, A. H. Behavioral responses to stress are intact in CRF-deficient mice. Brain Res. 845, 14–20 (1999).

  29. 29.

    Miklós, I. H. & Kovács, K. J. GABAergic innervation of corticotropin-releasing hormone (CRH)-secreting parvocellular neurons and its plasticity as demonstrated by quantitative immunoelectron microscopy. Neuroscience 113, 581–592 (2002).

  30. 30.

    Cole, R. L. & Sawchenko, P. E. Neurotransmitter regulation of cellular activation and neuropeptide gene expression in the paraventricular nucleus of the hypothalamus. J. Neurosci. 22, 959–969 (2002).

  31. 31.

    Hewitt, S. A., Wamsteeker, J. I., Kurz, E. U. & Bains, J. S. Altered chloride homeostasis removes synaptic inhibitory constraint of the stress axis. Nat. Neurosci. 12, 438–443 (2009).

  32. 32.

    Walker, L. C., Cornish, L. C., Lawrence, A. J. & Campbell, E. J. The effect of acute or repeated stress on the corticotropin releasing factor system in the CRH-IRES-Cre mouse: a validation study. Neuropharmacology https://doi.org/10.1016/j.neuropharm.2018.09.037 (2018).

  33. 33.

    Falkner, A. L., Grosenick, L., Davidson, T. J., Deisseroth, K. & Lin, D. Hypothalamic control of male aggression-seeking behavior. Nat. Neurosci. 19, 596–604 (2016).

  34. 34.

    Lin, D. et al. Functional identification of an aggression locus in the mouse hypothalamus. Nature 470, 221–226 (2011).

  35. 35.

    García-Lecumberri, C. & Ambrosio, E. Role of corticotropin-releasing factor in forced swimming test. Eur. J. Pharmacol. 343, 17–26 (1998).

  36. 36.

    Gao, V., Vitaterna, M. H. & Turek, F. W. Validation of video motion-detection scoring of forced swim test in mice. J. Neurosci. Methods 235, 59–64 (2014).

  37. 37.

    Saraiva, L. R. et al. Combinatorial effects of odorants on mouse behavior. Proc. Natl Acad. Sci. USA 113, E3300–E3306 (2016).

  38. 38.

    Rubinow, M. J., Hagerbaumer, D. A. & Juraska, J. M. The food-conditioned place preference task in adolescent, adult and aged rats of both sexes. Behav. Brain Res. 198, 263–266 (2009).

  39. 39.

    Cusulin, J. I. W., Füzesi, T., Watts, A. G. & Bains, J. S. Characterization of corticotropin-releasing hormone neurons in the paraventricular nucleus of the hypothalamus of Crh-IRES-Cre mutant mice. PLoS One 8, e64943 (2013).

  40. 40.

    Park, S. G. et al. Medial preoptic circuit induces hunting-like actions to target objects and prey. Nat. Neurosci. 21, 364–372 (2018).

  41. 41.

    Fang, Y. Y., Yamaguchi, T., Song, S. C., Tritsch, N. X. & Lin, D. A hypothalamic midbrain pathway essential for driving maternal behaviors. Neuron 98, 192–207.e10 (2018).

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We are grateful to K. Narasimhan, L. Vendruscolo, and members of the Suh laboratory for critical comments on the manuscript. We also thank G. Schwartz, S. Han, J. Kim, and A. Watts for discussions of this work. We appreciate W. Jung for assisting with immunohistochemistry and data analysis, and the laboratory of the late W. Vale for providing an aliquot of anti-CRF antibody. This work is supported by the TJ Park Science Fellowship of the POSCO TJ Park Foundation and the KAIST Innovative Doctoral Research Fellowship to J.K., NIH grants (R01MH101377 to D.L. and R01DK106636 to G.S.B.S.), and KAIST Chancellor’s fund to G.S.B.S. and J.W.S.

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Author notes

  1. These authors contributed equally: S. Lee, Y.-Y. Fang.


  1. Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea

    • Jineun Kim
    • , Seongju Lee
    • , Anna Shin
    • , Seahyung Park
    • , Shreelatha Bhat
    • , Daesoo Kim
    • , Jong-Woo Sohn
    •  & Greg S. B. Suh
  2. Neuroscience Institute, New York University School of Medicine, New York, NY, USA

    • Yi-Ya Fang
    • , Koichi Hashikawa
    • , Dayu Lin
    •  & Greg S. B. Suh
  3. Department of Psychiatry, New York University School of Medicine, New York, NY, USA

    • Yi-Ya Fang
    • , Koichi Hashikawa
    •  & Dayu Lin
  4. Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY, USA

    • Greg S. B. Suh
  5. Department of Cell Biology, New York University School of Medicine, New York, NY, USA

    • Greg S. B. Suh


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G.S.B.S., D.L., and J.K. conceived the project, designed the experiments, and interpreted the results. G.S.B.S. wrote the manuscript with D.L., J.K. and S.L. J.K. performed the experiments with assistance from S.L., Y.-Y.F., A.S., S.P., S.B., and D.K. J.K. and D.L. analyzed the calcium imaging data. Y.-Y.F., K.H., and D.L. made it possible for J.K. to carry out fiber photometry recordings.

Competing interests

The authors declare no competing interests.

Corresponding authors

Correspondence to Dayu Lin or Greg S. B. Suh.

Integrated supplementary information

  1. Supplementary Figure 1 PVN-CRFGCaMP6 signal is increased by the onset and is decreased by the offset of exposure to aversive stimuli.

    ac, PETH plot (upper) and heat map (bottom) from a representative animal shown in Fig. 1d to the onset of TRT (a), in Fig. 1f to the onset of overhead object (b), and in Fig. 1f’ to the onset of visual looming disk (c). eg, PETH plot (upper) and heat map (bottom) for population activity across animals to the offset of TRT (e), the offset of overhead object (f), and the offset of visual looming disk (g). d,h, PETH plot (upper) and heat map (bottom) from a representative animal shown in Fig. 1f’, aligned to the onset of flight behavior (d) and population activity across animals (h). Data are presented as mean (solid line) ± s.e.m (shaded area). Related to Fig. 1.

  2. Supplementary Figure 2 The changes in ∆F/F of PVN-CRFGFP signal in response to environmental stimuli were not due to a movement artifact.

    a,b, Left: a representative trace illustrating no change in the normalized PVN-CRFGFP signal during FST (a) or TRT (b). Right: bar graph illustrating average ΔF/F of PVN-CRFGFP signals during FST (a) and TRT (b) (N = 5 mice). This control experiment was conducted to ascertain the validity of increased PVN CRF neuronal activity in response to stressors that caused movements. Data are presented as mean ± s.e.m. Related to Fig. 1.

  3. Supplementary Figure 3 PVN-CRFGCaMP6 signal is increased by a flying object presented from above, but not by objects presented from either side or from below.

    a, Schematic for a ‘flying’ object from above and side (left). Bar graph showing average ΔF/F of PVN-CRFGCaMP6 in mice that were exposed to the flying object from above (N = 7 mice) and side (N = 4 mice) (right). b, Schematic for a looming shadow disk (left). Bar graph showing average ΔF/F of PVN-CRFGCaMP6 in mice that were exposed to the disk from above, side, or bottom (right) (N = 6 mice). Mann–Whitney two-tailed U-test in a and paired two-tailed t-test in b. *P < 0.05, **P < 0.01. See Supplementary Table 1 for detailed description of statistics for this figure and subsequent figures. Data are presented as mean ± s.e.m. Related to Fig. 1.

  4. Supplementary Figure 4 PVN-CRFGCaMP6 signals and c-Fos induction in PVN CRF neurons after periods of starvation.

    a,b, Bar graphs summarizing the levels of the baseline intracellular calcium (a) and the frequencies of calcium transients in PVN-CRFGCaMP6 mice when fed, 22 h fasted, and refed with chow (b) (N = 7). c,d, A representative confocal image of PVN in mice fasted for 22 h (c) or for 9 h (d) immunostained with anti-CRF (red) and anti-c-Fos (green) antibodies. Similar results were independently obtained in three other mice for 22 h fasted and 3 mice for 9 h fasted. Scale bar, 50 μm. One-way ANOVA test with Holm–Sidak post hoc analysis. *P < 0.05, **P < 0.01. Data are presented as mean ± s.e.m. Related to Fig. 2.

  5. Supplementary Figure 5 PVN-CRFGCaMP6 signals decrease when mice investigate and consume food for the first time.

    af, PETH plot of population activity across animals, for the first bout and all other bouts to the onset of investigation of a mesh-covered cup with object (a), chow pellet (c), or peanut butter (e); investigation of object (b); and consumption of chow pellet (d) and peanut butter (f) in fed (left) or fasted (right) mice (N = 11). Data are presented as mean ± s.e.m. Related to Fig. 2.

  6. Supplementary Figure 6 The frequency of calcium transients decreases during the presentation of food or a pup.

    a,c, Representative traces with denoted calcium transients (red dots) of raw PVN-CRFGCaMP6 signals in a mouse that was fed (left) or fasted (right) when exposed to object or chow (a); and in a male (left) or a female (right) when exposed to a pup (c). Blue line denotes the moment at which chow (a) or a pup (c) was introduced. b,d, Bar graph summarizing the frequency of calcium transients of PVN-CRFGCaMP6 signals during the presentation of object and chow (N = 11 mice) (b); before and after pups were introduced to male (N = 12 mice) or female mice (N = 8 mice) (d). One-way ANOVA test with Holm–Sidak post hoc analysis in b and paired two-tailed t-test in d. *P < 0.05, **P < 0.01. Data are presented as mean ± s.e.m. Related to Figs. 1 and 3.

  7. Supplementary Figure 7 PVN-CRFGCaMP6 signals decrease when female mice approach and interact with pups for the first time.

    a,b, PETH plots for population activity across animals to the onset of approach to pups (a) and interaction with pups (b) by male (left, N = 12) or female (right, N = 8) mice. First bout and all other bouts are overlaid. c, Representative traces illustrating no change in the normalized PVN-CRFGCaMP6 signals in a male (left) or a female (right) during the presence of a fake animal. Shaded bars depict the epochs during which the recorded mouse approached toward the fake animal (light gray) and interaction with the fake animal (light purple). d,e, PETH plots for population activity across animals to the onset of approach to fake animals (d) and interaction with fake animals (e) by male (left, N = 12) and female (right, N = 7) mice. Data are presented as mean ± s.e.m. Related to Fig. 3.

  8. Supplementary Figure 8 PVN-CRFGCaMP6 signals of recorded female mice while being aggressed on by CD1 female aggressor.

    a, A representative trace illustrating acute surge in PVN-CRFGCaMP6 signal in a female mouse to the onsets of being investigated (light gray) or being attacked (red) by a female aggressor, and being introduced or removed from the arena (gray). b, PETH plot across trials of being attacked from the representative trace (n = 10 trials). Similar results were independently obtained by four other female mice. Data are presented as mean ± s.e.m. Related to Fig. 3.

  9. Supplementary Figure 9 PVN-CRFGCaMP6 signals of recorded mice while interacting with CD1 aggressor.

    A population PETH plot from recorded mice during investigation with aggressive intruders that does not involve attack (left) and bar graph that measures the changes in ∆F/F of PVN-CRFGCaMP6 signal (right) (N = 5 mice). Paired two-tailed t-test. P = 0.11. Data are presented as mean ± s.e.m. Related to Fig. 3.

  10. Supplementary Figure 10 Optogenetic activation of PVN CRF neurons of food-restricted mice reduces preference for food and for food-paired chamber.

    ae, Time spent in eating normalized to time spent in the food-paired chamber (%) (a), the number of eating bouts (b), the duration of eating bout (c), probability of initiating eating when mice are in the food-paired chamber (%) (d), and cumulative bouts from days 3, 4, and 5 (e) in PVN-CRFChR2 mice (blue, N = 12) and control PVN-CRFeYFP (gray, N = 9) mice. Two-way ANOVA test with Holm–Sidak post hoc analysis. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data are presented as mean ± s.e.m. Related to Fig. 6.

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