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GABA and glutamate neurons in the VTA regulate sleep and wakefulness

Abstract

We screened for novel circuits in the mouse brain that promote wakefulness. Chemogenetic activation experiments and electroencephalogram recordings pointed to glutamatergic/nitrergic (NOS1) and GABAergic neurons in the ventral tegmental area (VTA). Activating glutamatergic/NOS1 neurons, which were wake- and rapid eye movement (REM) sleep-active, produced wakefulness through projections to the nucleus accumbens and the lateral hypothalamus. Lesioning the glutamate cells impaired the consolidation of wakefulness. By contrast, activation of GABAergic VTA neurons elicited long-lasting non-rapid-eye-movement-like sleep resembling sedation. Lesioning these neurons produced an increase in wakefulness that persisted for at least 4 months. Surprisingly, these VTA GABAergic neurons were wake- and REM sleep-active. We suggest that GABAergic VTA neurons may limit wakefulness by inhibiting the arousal-promoting VTA glutamatergic and/or dopaminergic neurons and through projections to the lateral hypothalamus. Thus, in addition to its contribution to goal- and reward-directed behaviors, the VTA has a role in regulating sleep and wakefulness.

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The datasets generated during and/or analyzed during the current study are available from the corresponding authors on reasonable request.

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References

  1. 1.

    Scammell, T. E., Arrigoni, E. & Lipton, J. O. Neural circuitry of wakefulness and sleep. Neuron 93, 747–765 (2017).

  2. 2.

    Weber, F. & Dan, Y. Circuit-based interrogation of sleep control. Nature 538, 51–59 (2016).

  3. 3.

    Saper, C. B. & Fuller, P. M. Wake–sleep circuitry: an overview. Curr. Opin. Neurobiol. 44, 186–192 (2017).

  4. 4.

    Gent, T. C., Bassetti, C. & Adamantidis, A. R. Sleep–wake control and the thalamus. Curr. Opin. Neurobiol. 52, 188–197 (2018).

  5. 5.

    Eban-Rothschild, A., Rothschild, G., Giardino, W. J., Jones, J. R. & de Lecea, L. VTA dopaminergic neurons regulate ethologically relevant sleep–wake behaviors. Nat. Neurosci. 19, 1356–1366 (2016).

  6. 6.

    Yu, X. et al. Wakefulness is governed by GABA and histamine cotransmission. Neuron 87, 164–178 (2015).

  7. 7.

    Cho, J. R. et al. Dorsal raphe dopamine neurons modulate arousal and promote wakefulness by salient stimuli. Neuron 94, 1205–1219.e8 (2017).

  8. 8.

    Oishi, Y. et al. Activation of ventral tegmental area dopamine neurons produces wakefulness through dopamine D2-like receptors in mice. Brain Struct. Funct. 222, 2907–2915 (2017).

  9. 9.

    Kosse, C., Schöne, C., Bracey, E. & Burdakov, D. Orexin-driven GAD65 network of the lateral hypothalamus sets physical activity in mice. Proc. Natl Acad. Sci. USA 114, 4525–4530 (2017).

  10. 10.

    Schöne, C., Apergis-Schoute, J., Sakurai, T., Adamantidis, A. & Burdakov, D. Coreleased orexin and glutamate evoke nonredundant spike outputs and computations in histamine neurons. Cell Rep. 7, 697–704 (2014).

  11. 11.

    Herrera, C. G. et al. Hypothalamic feedforward inhibition of thalamocortical network controls arousal and consciousness. Nat. Neurosci. 19, 290–298 (2016).

  12. 12.

    Anaclet, C. et al. Basal forebrain control of wakefulness and cortical rhythms. Nat. Commun. 6, 8744 (2015).

  13. 13.

    Xu, M. et al. Basal forebrain circuit for sleep–wake control. Nat. Neurosci. 18, 1641–1647 (2015).

  14. 14.

    Pedersen, N. P. et al. Supramammillary glutamate neurons are a key node of the arousal system. Nat. Commun. 8, 1405 (2017).

  15. 15.

    Venner, A., Anaclet, C., Broadhurst, R. Y., Saper, C. B. & Fuller, P. M. A novel population of wake-promoting GABAergic neurons in the ventral lateral hypothalamus. Curr. Biol. 26, 2137–2143 (2016).

  16. 16.

    Gent, T. C., Bandarabadi, M., Herrera, C. G. & Adamantidis, A. R. Thalamic dual control of sleep and wakefulness. Nat. Neurosci. 21, 974–984 (2018).

  17. 17.

    Ren, S. et al. The paraventricular thalamus is a critical thalamic area for wakefulness. Science 362, 429–434 (2018).

  18. 18.

    Weber, F. et al. Regulation of REM and non-REM sleep by periaqueductal GABAergic neurons. Nat. Commun. 9, 354 (2018).

  19. 19.

    Chung, S. et al. Identification of preoptic sleep neurons using retrograde labelling and gene profiling. Nature 545, 477–481 (2017).

  20. 20.

    Sherin, J. E., Shiromani, P. J., McCarley, R. W. & Saper, C. B. Activation of ventrolateral preoptic neurons during sleep. Science 271, 216–219 (1996).

  21. 21.

    Harding, E. C. et al. A neuronal hub binding sleep initiation and body cooling in response to a warm external stimulus. Curr. Biol. 28, 2263–2273.e4 (2018).

  22. 22.

    Anaclet, C. et al. The GABAergic parafacial zone is a medullary slow wave sleep-promoting center. Nat. Neurosci. 17, 1217–1224 (2014).

  23. 23.

    Uygun, D. S. et al. Bottom-up versus top-down induction of sleep by zolpidem acting on histaminergic and neocortex neurons. J. Neurosci. 36, 11171–11184 (2016).

  24. 24.

    Morales, M. & Margolis, E. B. Ventral tegmental area: cellular heterogeneity, connectivity and behaviour. Nat. Rev. Neurosci. 18, 73–85 (2017).

  25. 25.

    Russo, S. J. & Nestler, E. J. The brain reward circuitry in mood disorders. Nat. Rev. Neurosci. 14, 609–625 (2013).

  26. 26.

    Lüscher, C. The emergence of a circuit model for addiction. Annu. Rev. Neurosci. 39, 257–276 (2016).

  27. 27.

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

  28. 28.

    Hung, L. W. et al. Gating of social reward by oxytocin in the ventral tegmental area. Science 357, 1406–1411 (2017).

  29. 29.

    Hnasko, T. S., Hjelmstad, G. O., Fields, H. L. & Edwards, R. H. Ventral tegmental area glutamate neurons: electrophysiological properties and projections. J. Neurosci. 32, 15076–15085 (2012).

  30. 30.

    Taylor, S. R. et al. GABAergic and glutamatergic efferents of the mouse ventral tegmental area. J. Comp. Neurol. 522, 3308–3334 (2014).

  31. 31.

    Creed, M. C., Ntamati, N. R. & Tan, K. R. VTA GABA neurons modulate specific learning behaviors through the control of dopamine and cholinergic systems. Front. Behav. Neurosci. 8, 8 (2014).

  32. 32.

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

  33. 33.

    Taylor, N. E. et al. Optogenetic activation of dopamine neurons in the ventral tegmental area induces reanimation from general anesthesia. Proc. Natl Acad. Sci. USA 113, 12826–12831 (2016).

  34. 34.

    Paul, E. J. et al. nNOS-expressing neurons in the ventral tegmental area and substantia nigra pars compacta. eNeuro 5, e0381-18 (2018).

  35. 35.

    Qi, J. et al. VTA glutamatergic inputs to nucleus accumbens drive aversion by acting on GABAergic interneurons. Nat. Neurosci. 19, 725–733 (2016).

  36. 36.

    Yamaguchi, T., Wang, H. L., Li, X., Ng, T. H. & Morales, M. Mesocorticolimbic glutamatergic pathway. J. Neurosci. 31, 8476–8490 (2011).

  37. 37.

    Gomez, J. L. et al. Chemogenetics revealed: DREADD occupancy and activation via converted clozapine. Science 357, 503–507 (2017).

  38. 38.

    van Zessen, R., Phillips, J. L., Budygin, E. A. & Stuber, G. D. Activation of VTA GABA neurons disrupts reward consumption. Neuron 73, 1184–1194 (2012).

  39. 39.

    Fifel, K., Meijer, J. H. & Deboer, T. Circadian and homeostatic modulation of multi-unit activity in midbrain dopaminergic structures. Sci. Rep. 8, 7765 (2018).

  40. 40.

    Blanco-Centurion, C., Gerashchenko, D. & Shiromani, P. J. Effects of saporin-induced lesions of three arousal populations on daily levels of sleep and wake. J. Neurosci. 27, 14041–14048 (2007).

  41. 41.

    Root, D. H. et al. Single rodent mesohabenular axons release glutamate and GABA. Nat. Neurosci. 17, 1543–1551 (2014).

  42. 42.

    Nieh, E. H. et al. Inhibitory input from the lateral hypothalamus to the ventral tegmental area disinhibits dopamine neurons and promotes behavioral activation. Neuron 90, 1286–1298 (2016).

  43. 43.

    Faget, L. et al. Afferent inputs to neurotransmitter-defined cell types in the ventral tegmental area. Cell Rep. 15, 2796–2808 (2016).

  44. 44.

    Gelegen, C. et al. Excitatory pathways from the lateral habenula enable propofol-induced sedation. Curr. Biol. 28, 580–587.e5 (2018).

  45. 45.

    Jhou, T. C., Fields, H. L., Baxter, M. G., Saper, C. B. & Holland, P. C. The rostromedial tegmental nucleus (RMTg), a GABAergic afferent to midbrain dopamine neurons, encodes aversive stimuli and inhibits motor responses. Neuron 61, 786–800 (2009).

  46. 46.

    Qiu, M. H., Vetrivelan, R., Fuller, P. M. & Lu, J. Basal ganglia control of sleep–wake behavior and cortical activation. Eur. J. Neurosci. 31, 499–507 (2010).

  47. 47.

    Qiu, M. H. et al. The role of nucleus accumbens core/shell in sleep–wake regulation and their involvement in modafinil-induced arousal. PLoS One 7, e45471 (2012).

  48. 48.

    Morairty, S. R. et al. A role for cortical nNOS/NK1 neurons in coupling homeostatic sleep drive to EEG slow wave activity. Proc. Natl Acad. Sci. USA 110, 20272–20277 (2013).

  49. 49.

    Geisler, S., Derst, C., Veh, R. W. & Zahm, D. S. Glutamatergic afferents of the ventral tegmental area in the rat. J. Neurosci. 27, 5730–5743 (2007).

  50. 50.

    Beier, K. T. et al. Circuit architecture of VTA dopamine neurons revealed by systematic input-output mapping. Cell 162, 622–634 (2015).

  51. 51.

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

  52. 52.

    Leshan, R. L. et al. Leptin action through hypothalamic nitric oxide synthase-1-expressing neurons controls energy balance. Nat. Med. 18, 820–823 (2012).

  53. 53.

    Taniguchi, H. et al. A resource of Cre driver lines for genetic targeting of GABAergic neurons in cerebral cortex. Neuron 71, 995–1013 (2011).

  54. 54.

    Hippenmeyer, S. et al. A developmental switch in the response of DRG neurons to ETS transcription factor signaling. PLoS Biol. 3, e159 (2005).

  55. 55.

    Krashes, M. J. et al. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J. Clin. Invest. 121, 1424–1428 (2011).

  56. 56.

    Yang, C. F. et al. Sexually dimorphic neurons in the ventromedial hypothalamus govern mating in both sexes and aggression in males. Cell 153, 896–909 (2013).

  57. 57.

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

  58. 58.

    Klugmann, M. et al. AAV-mediated hippocampal expression of short and long Homer 1 proteins differentially affect cognition and seizure activity in adult rats. Mol. Cell. Neurosci. 28, 347–360 (2005).

  59. 59.

    Murray, A. J. et al. Parvalbumin-positive CA1 interneurons are required for spatial working but not for reference memory. Nat. Neurosci. 14, 297–299 (2011).

  60. 60.

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

  61. 61.

    Klapoetke, N. C. et al. Independent optical excitation of distinct neural populations. Nat. Methods 11, 338–346 (2014).

  62. 62.

    Jennings, J. H. et al. Distinct extended amygdala circuits for divergent motivational states. Nature 496, 224–228 (2013).

  63. 63.

    Anisimov, V. N. et al. Reconstruction of vocal interactions in a group of small songbirds. Nat. Methods 11, 1135–1137 (2014).

  64. 64.

    Flusberg, B. A. et al. High-speed, miniaturized fluorescence microscopy in freely moving mice. Nat. Methods 5, 935–938 (2008).

  65. 65.

    Groessl, F. et al. Dorsal tegmental dopamine neurons gate associative learning of fear. Nat. Neurosci. 21, 952–962 (2018).

  66. 66.

    Chen, K. S. et al. A hypothalamic switch for REM and non-REM sleep. Neuron 97, 1168–1176.e4 (2018).

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Acknowledgements

We thank M. Ungless (Faculty of Medicine, Imperial College London) for comments on the manuscript. Our work was supported by the Wellcome Trust (107839/Z/15/Z, N.P.F. and 107841/Z/15/Z, W.W), the UK Dementia Research Institute (W.W. and N.P.F.), the Funds for International Cooperation and Exchange of the National Natural Science Foundation of China (grant no. 81620108012, H.D. and N.P.F.), the China Scholarship Council (Y.M.), a Rubicon Fellowship (019.161LW.010) from the Netherlands Organization for Scientific Research (W.B.), an Imperial College Schrödinger Scholarship (G.M.), and an Imperial College Junior Research Fellowship (J.J.H.). D.B. and J.J.H. were also supported by The Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001055), the Medical Research Council (FC001055), and the Wellcome Trust (FC001055). The Facility for Imaging by Light Microscopy at Imperial College London is in part supported by funding from the Wellcome Trust (grant no. 104931/Z/14/Z) and BBSRC (grant no. BB/L015129/1).

Author information

N.P.F. and W.W. conceived and, with X.Y. and H.D., designed the experiments. X.Y., W.L., Y.M., K.T., J.J.H., E.C.H., W.B., G.M., D.W., L.L., J.G., M.C., Y.L., R.Y., D.B., and Q. Y. performed the experiments and/or data analysis. A.L.V. provided the Neurologgers. N.P.F. and W.W. contributed to the data analysis and with H.D. supervised the project. N.P.F., X.Y., and W.W. wrote the paper.

Competing interests

The authors declare no competing interests.

Correspondence to Hailong Dong or Nicholas P. Franks or William Wisden.

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Fig. 1: Chemogenetic mapping for novel glutamatergic areas in the PH/MB that promote wakefulness identifies the VTA.
Fig. 2: VTAVglut2 neurons consolidate wakefulness and are selectively wake- and REM sleep-active.
Fig. 3: VTAVglut2 neurons promote wakefulness by their projections to the LH and NAc.
Fig. 4: VTAVglut2 and VTANos1 neurons promote wakefulness.
Fig. 5: Excitation of GABAergic neurons in the VTA induces sleep and their inhibition produces continuous wakefulness.
Fig. 6: VTAVgat neurons inhibit wakefulness; lesioning of VTAVgat neurons produces extended wakefulness, but VTAVgat neurons are selectively wake- and REM-active.
Fig. 7: VTAVgat neurons limit wakefulness in part by locally inhibiting dopamine and Vglut2 neurons in the VTA.
Fig. 8: VTAVgat neurons inhibit wakefulness in part via projections to the LH.