Zona incerta GABAergic neurons integrate prey-related sensory signals and induce an appetitive drive to promote hunting


The neural substrates for predatory hunting, an evolutionarily conserved appetitive behavior, remain largely undefined. Photoactivation of zona incerta (ZI) GABAergic neurons strongly promotes hunting of both live and artificial prey. Conversely, photoinhibition of these neurons or deletion of their GABA function severely impairs hunting. Here electrophysiological recordings reveal that ZI neurons integrate prey-related multisensory signals and discriminate prey from non-prey targets. Visual or whisker sensory deprivation reduces calcium responses induced by prey introduction and attack and impair hunting. ZI photoactivation largely corrects the hunting impairment caused by sensory deprivations. Motivational and reinforcing assays reveal that ZI photoactivation is associated with a strong appetitive drive, causing repetitive self-stimulatory behaviors. These ZI neurons project to the periaqueductal gray matter to induce hunting and motivation. Thus, we have delineated the function of ZI GABAergic neurons in hunting, which integrates prey-related sensory signals into prey detection and attack and induces a strong appetitive motivational drive.

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Fig. 1: Photoactivation of ZIm GABAergic neurons promotes predatory hunting.
Fig. 2: Requirement of ZIm GABAergic neurons and GABA in predatory hunting.
Fig. 3: ZIm neurons are sensitive to hunting-related signals.
Fig. 4: ZI GABAergic neurons incorporate sensory signals into hunting.
Fig. 5: ZIm GABAergic neurons induce an appetitive motivational drive.
Fig. 6: ZIm GABAergic projections to the PAG promote hunting.

Data availability

The data that support the findings of this study are available from the corresponding authors upon reasonable request.

Code availability

The code that supports the findings of this study is available from the corresponding authors upon reasonable request.


  1. 1.

    Volkmann, J., Daniels, C. & Witt, K. Neuropsychiatric effects of subthalamic neurostimulation in Parkinson disease. Nat. Rev. Neurol. 6, 487–498 (2010).

    CAS  Article  Google Scholar 

  2. 2.

    Morgan, J. C. et al. Self-stimulatory behavior associated with deep brain stimulation in Parkinson’s disease. Mov. Disord. 21, 283–285 (2006).

    Article  Google Scholar 

  3. 3.

    Kim, H. J., Jeon, B. S. & Paek, S. H. Nonmotor symptoms and subthalamic deep brain stimulation in Parkinson’s disease. J. Mov. Disord. 8, 83–91 (2015).

    Article  Google Scholar 

  4. 4.

    Caire, F., Ranoux, D., Guehl, D., Burbaud, P. & Cuny, E. A systematic review of studies on anatomical position of electrode contacts used for chronic subthalamic stimulation in Parkinson’s disease. Acta Neurochir. 155, 1647–1654 (2013).

    Article  Google Scholar 

  5. 5.

    Zhang, X. & van den Pol, A. N. Rapid binge-like eating and body weight gain driven by zona incerta GABA neuron activation. Science 356, 853–859 (2017).

    CAS  Article  Google Scholar 

  6. 6.

    Liu, K. et al. Lhx6-positive GABA-releasing neurons of the zona incerta promote sleep. Nature 548, 582–587 (2017).

    CAS  Article  Google Scholar 

  7. 7.

    Roger, M. & Cadusseau, J. Afferents to the zona incerta in the rat: a combined retrograde and anterograde study. J. Comp. Neurol. 241, 480–492 (1985).

    CAS  Article  Google Scholar 

  8. 8.

    Power, B. D., Leamey, C. A. & Mitrofanis, J. Evidence for a visual subsector within the zona incerta. Vis. Neurosci. 18, 179–186 (2001).

    CAS  Article  Google Scholar 

  9. 9.

    Chometton, S. et al. The rostromedial zona incerta is involved in attentional processes while adjacent LHA responds to arousal: c-Fos and anatomical evidence. Brain Struct. Funct. 222, 2507–2525 (2017).

    CAS  Article  Google Scholar 

  10. 10.

    Tait, D. S., Phillips, J. M., Blackwell, A. D. & Brown, V. J. Effects of lesions of the subthalamic nucleus/zona incerta area and dorsomedial striatum on attentional set-shifting in the rat. Neuroscience 345, 287–296 (2017).

    CAS  Article  Google Scholar 

  11. 11.

    Perier, C., Tremblay, L., Feger, J. & Hirsch, E. C. Behavioral consequences of bicuculline injection in the subthalamic nucleus and the zona incerta in rat. J. Neurosci. 22, 8711–8719 (2002).

    CAS  Article  Google Scholar 

  12. 12.

    Murer, M. G. & Pazo, J. H. Circling behaviour induced by activation of GABAA receptors in the subthalamic nucleus. Neuroreport 4, 1219–1222 (1993).

    CAS  Article  Google Scholar 

  13. 13.

    Milner, K. L. & Mogenson, G. J. Electrical and chemical activation of the mesencephalic and subthalamic locomotor regions in freely moving rats. Brain Res. 452, 273–285 (1988).

    CAS  Article  Google Scholar 

  14. 14.

    Supko, D. E., Uretsky, N. J. & Wallace, L. J. Activation of AMPA/kainic acid glutamate receptors in the zona incerta stimulates locomotor activity. Brain Res. 564, 159–163 (1991).

    CAS  Article  Google Scholar 

  15. 15.

    Huang, Y. H. & Mogenson, G. J. Differential effects of incertal and hypothalamic lesions on food and water intake. Exp. Neurol. 43, 276–280 (1974).

    CAS  Article  Google Scholar 

  16. 16.

    Walsh, L. L., Halaris, A. E., Grossman, L. & Grossman, S. P. Some biochemical effects of zona incerta lesions that interfere with the regulation of water intake. Pharm. Biochem. Behav. 7, 351–356 (1977).

    CAS  Article  Google Scholar 

  17. 17.

    Mitofanis, J. Evidence for an auditory subsector within the zona incerta of rats. Anat. Embryol. 205, 453–462 (2002).

    Article  Google Scholar 

  18. 18.

    Shaw, V. & Mitrofanis, J. Anatomical evidence for somatotopic maps in the zona incerta of rats. Anat. Embryol. 206, 119–130 (2002).

    CAS  Article  Google Scholar 

  19. 19.

    Mitrofanis, J. Some certainty for the “zone of uncertainty”? exploring the function of the zona incerta. Neuroscience 130, 1–15 (2005).

    CAS  Article  Google Scholar 

  20. 20.

    Power, B. D. & Mitrofanis, J. Zona incerta: substrate for contralateral interconnectivity in the thalamus of rats. J. Comp. Neurol. 436, 52–63 (2001).

    CAS  Article  Google Scholar 

  21. 21.

    Power, B. D. & Mitrofanis, J. Evidence for extensive inter-connections within the zona incerta in rats. Neurosci. Lett. 267, 9–12 (1999).

    CAS  Article  Google Scholar 

  22. 22.

    Watson, G. D., Smith, J. B. & Alloway, K. D. The zona incerta regulates communication between the superior colliculus and the posteromedial thalamus: implications for thalamic interactions with the dorsolateral striatum. J. Neurosci. 35, 9463–9476 (2015).

    CAS  Article  Google Scholar 

  23. 23.

    Power, B. D. & Mitrofanis, J. Ultrastructure of afferents from the zona incerta to the posterior and parafascicular thalamic nuclei of rats. J. Comp. Neurol. 451, 33–44 (2002).

    Article  Google Scholar 

  24. 24.

    Kolmac, C. I., Power, B. D. & Mitrofanis, J. Patterns of connections between zona incerta and brainstem in rats. J. Comp. Neurol. 396, 544–555 (1998).

    CAS  Article  Google Scholar 

  25. 25.

    Power, B. D., Kolmac, C. I. & Mitrofanis, J. Evidence for a large projection from the zona incerta to the dorsal thalamus. J. Comp. Neurol. 404, 554–565 (1999).

    CAS  Article  Google Scholar 

  26. 26.

    Won, R. Motivation: A Biobehavioural Approach (Cambridge Univ. Press, 2000).

  27. 27.

    Furigo, I. C. et al. The role of the superior colliculus in predatory hunting. Neuroscience 165, 1–15 (2010).

    CAS  Article  Google Scholar 

  28. 28.

    Butler, K. Predatory behavior in laboratory mice: strain and sex comparisons. J. Comp. Physiol. Psychol. 85, 243–249 (1973).

    CAS  Article  Google Scholar 

  29. 29.

    Han, W. et al. Integrated control of predatory hunting by the central nucleus of the amygdala. Cell 168, 311–324.e318 (2017).

    CAS  Article  Google Scholar 

  30. 30.

    Hoy, J. L., Yavorska, I., Wehr, M. & Niell, C. M. Vision drives accurate approach behavior during prey capture in laboratory mice. Curr. Biol. 26, 3046–3052 (2016).

    CAS  Article  Google Scholar 

  31. 31.

    Favaro, P. D. et al. The influence of vibrissal somatosensory processing in rat superior colliculus on prey capture. Neuroscience 176, 318–327 (2011).

    CAS  Article  Google Scholar 

  32. 32.

    Mota-Ortiz, S. R. et al. The periaqueductal gray as a critical site to mediate reward seeking during predatory hunting. Behav. Brain Res. 226, 32–40 (2012).

    Article  Google Scholar 

  33. 33.

    Comoli, E., Ribeiro-Barbosa, E. R. & Canteras, N. S. Predatory hunting and exposure to a live predator induce opposite patterns of Fos immunoreactivity in the PAG. Behav. Brain Res. 138, 17–28 (2003).

    CAS  Article  Google Scholar 

  34. 34.

    Comoli, E., Ribeiro-Barbosa, E. R., Negrao, N., Goto, M. & Canteras, N. S. Functional mapping of the prosencephalic systems involved in organizing predatory behavior in rats. Neuroscience 130, 1055–1067 (2005).

    CAS  Article  Google Scholar 

  35. 35.

    Li, Y. et al. Hypothalamic circuits for predation and evasion. Neuron 97, 911–924.e5 (2018).

    CAS  Article  Google Scholar 

  36. 36.

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

    CAS  Article  Google Scholar 

  37. 37.

    Lin, J. Y., Lin, M. Z., Steinbach, P. & Tsien, R. Y. Characterization of engineered channelrhodopsin variants with improved properties and kinetics. Biophys. J. 96, 1803–1814 (2009).

    CAS  Article  Google Scholar 

  38. 38.

    Vong, L. et al. Leptin action on GABAergic neurons prevents obesity and reduces inhibitory tone to POMC neurons. Neuron 71, 142–154 (2011).

    CAS  Article  Google Scholar 

  39. 39.

    Govorunova, E. G., Sineshchekov, O. A., Janz, R., Liu, X. & Spudich, J. L. Natural light-gated anion channels: a family of microbial rhodopsins for advanced optogenetics. Science 349, 647–650 (2015).

    CAS  Article  Google Scholar 

  40. 40.

    Cui, Y. et al. A central amygdala–substantia innominata neural circuitry encodes aversive reinforcement signals. Cell Rep. 21, 1770–1782 (2017).

    CAS  Article  Google Scholar 

  41. 41.

    Chou, X. L. et al. Inhibitory gain modulation of defense behaviors by zona incerta. Nat. Commun. 9, 1151 (2018).

    Article  Google Scholar 

  42. 42.

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

    CAS  Article  Google Scholar 

  43. 43.

    Zhao, Z. D. et al. A hypothalamic circuit that controls body temperature. Proc. Natl Acad. Sci. USA 114, 2042–2047 (2017).

    CAS  Article  Google Scholar 

  44. 44.

    Shang, C. et al. A subcortical excitatory circuit for sensory-triggered predatory hunting in mice. Nat. Neurosci. https://doi.org/10.1038/s41593-019-0405-4 (2019).

  45. 45.

    Hodos, W. Progressive ratio as a measure of reward strength. Science 134, 943–944 (1961).

    CAS  Article  Google Scholar 

  46. 46.

    Pei, Y., Rogan, S. C., Yan, F. & Roth, B. L. Engineered GPCRs as tools to modulate signal transduction. Physiology 23, 313–321 (2008).

    CAS  Article  Google Scholar 

  47. 47.

    Tervo, D. G. et al. A designer AAV variant permits efficient retrograde access to projection neurons. Neuron 92, 372–382 (2016).

    CAS  Article  Google Scholar 

  48. 48.

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

    CAS  Article  Google Scholar 

  49. 49.

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

    CAS  Article  Google Scholar 

  50. 50.

    Bandler, R. J. Jr, Chi, C. C. & Flynn, J. P. Biting attack elicited by stimulation of the ventral midbrain tegmentum of cats. Science 177, 364–366 (1972).

    Article  Google Scholar 

  51. 51.

    Berridge, K. C. Motivation concepts in behavioral neuroscience. Physiol. Behav. 81, 179–209 (2004).

    CAS  Article  Google Scholar 

  52. 52.

    Paxinos, G. & Franklin, K. B. J. The Mouse in Sterotaxic Coordinates 3rd edn (Academic Press, 2008).

  53. 53.

    Douglass, A. M. et al. Central amygdala circuits modulate food consumption through a positive-valence mechanism. Nat. Neurosci. 20, 1384–1394 (2017).

    CAS  Article  Google Scholar 

  54. 54.

    Do-Monte, F. H., Minier-Toribio, A., Quinones-Laracuente, K., Medina-Colon, E. M. & Quirk, G. J. Thalamic regulation of sucrose seeking during unexpected reward omission. Neuron 94, 388–400.e384 (2017).

    CAS  Article  Google Scholar 

  55. 55.

    Wei, Y. C. et al. Medial preoptic area in mice is capable of mediating sexually dimorphic behaviors regardless of gender. Nat. Commun. 9, 279 (2018).

    Article  Google Scholar 

  56. 56.

    Tseng, W. T., Yen, C. T. & Tsai, M. L. A bundled microwire array for long-term chronic single-unit recording in deep brain regions of behaving rats. J. Neurosci. Methods 201, 368–376 (2011).

    Article  Google Scholar 

  57. 57.

    Guo, L., Walker, W. I., Ponvert, N. D., Penix, P. L. & Jaramillo, S. Stable representation of sounds in the posterior striatum during flexible auditory decisions. Nat. Commun. 9, 1534 (2018).

    Article  Google Scholar 

  58. 58.

    Chen, X. & Li, H. Ar control: an arduino-based comprehensive behavioral platform with real-time performance. Front. Behav. Neurosci. 11, 244 (2017).

    Article  Google Scholar 

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The authors thank the following individuals: M. Luo, H. Hu, C. Zhan, J. Liao, H. Zhu and Y. Zou for sharing reagents; X. Li and the Molecular Imaging Core Facility (MICF) of the School of Life Science and Technology, ShanghaiTech University, for microscopy imaging; Y. Xiong and the Molecular Cellular Core for slices and staining; all members of the Shen Lab and the “Shen Xian Hui (NPC)” Wechat group for valuable discussion. The authors also thank staff members from the Shanghai Model Organisms Center and Animal Facility at NFPS, Zhangjiang Lab, China. This study is funded by the National Natural Science Foundation of China (no. 91857104 and no. 31771169 to W.L.S.; no. 91432107 and no. 31671105 to H.L.), the Shanghai Municipal Education Commission (no. 2019-01-07-00-10-E00058 to W.L.S.), the Science Fund for Creative Research Group of China (no. 61721092 to H.L.), the Director Fund of the Wuhan National Laboratory for Optoelectronics (to H.L.), the Thousand Young Talents Program of China (to W.L.S.), the Shanghai Pujiang Talent Award (no. 2018X0302-101-01 to W.S.) and the ShanghaiTech University start-up fund (to W.L.S.).

Author information




Z.-D.Z., Zongming C., X.X. and M.H. performed most of the experiments. H.X., X.J., L.Q., C.S. and Y.Y. performed the behavioral evaluations. J.L. and X.N. performed the immunostaining. F.C. and Zijun C. performed the electrophysiology. Y.C. performed the multichannel recordings. Z.-D.Z., J.H., W.S., P.C., H.L. and W.L.S. designed the experiments. Z.-D. Z., Zongming C., M.H., H.L. and W.L.S. wrote the manuscript.

Corresponding authors

Correspondence to Haohong Li or Wei L. Shen.

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The authors declare no competing interests.

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Journal peer review information: Nature Neuroscience thanks Jennifer Hoy, Daesoo Kim and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Figures 1–25 and Supplementary Tables 1–3.

Reporting Summary

Supplementary Video 1

Home-cage behaviors. This video shows that photoactivation of zona incerta GABAergic neurons (ZImVgat-ChIEF) in food-supplied home-cages induces undirected-gnawing, biting objects, or binge-like eating. (Behavioral definitions: undirected gnawing, mice executed fictive-eating or gnawing uneatable bedding materials or feces; binge-like eating, mice rapidly bit and consumed food pellets; biting objects, mice bit on the food container or water nozzle.)

Supplementary Video 2

Photoactivation-induced bite attack. This video shows that one pulse of 100-ms photoactivation of zona incerta GABAergic neurons is sufficient to induce a bite attack on bedding materials.

Supplementary Video 3

ZI glutamatergic neurons did not promote hunting. This video shows that photoactivation of zona incerta glutamatergic neurons does not induce attacking, gnawing or eating of food or bedding materials.

Supplementary Video 4

Hunting of adult crickets. This video shows that adult crickets are very defensive and that photoactivation of zona incerta GABAergic neurons promotes hunting of adult crickets.

Supplementary Video 5

Attacking of artificial prey. This video shows that photoactivation of zona incerta GABAergic neurons elicits attacking of artificial prey.

Supplementary Video 6

Hunting behaviors of cricket-naive mice. This video shows that photoactivation of zona incerta GABAergic neurons increases attacking frequency in cricket-naive mice.

Supplementary Video 7

Photoactivation did not induce aggression. This video shows that photoactivation of zona incerta GABAergic neurons does not evoke attacking of male and female mice.

Supplementary Video 8

Photoinhibition interrupted hunting and eating. This video shows that photoinhibition of zona incerta GABAergic neurons in ZImVgat-GtACR mice abolishes cricket attacking, and eating.

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Zhao, Z., Chen, Z., Xiang, X. et al. Zona incerta GABAergic neurons integrate prey-related sensory signals and induce an appetitive drive to promote hunting. Nat Neurosci 22, 921–932 (2019). https://doi.org/10.1038/s41593-019-0404-5

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