Letter | Published:

Lhx6-positive GABA-releasing neurons of the zona incerta promote sleep

Nature 548, 582–587 (31 August 2017) | Download Citation

Subjects

  • A Corrigendum to this article was published on 27 September 2017

Abstract

Multiple populations of wake-promoting neurons have been characterized in mammals, but few sleep-promoting neurons have been identified1. Wake-promoting cell types include hypocretin and GABA (γ-aminobutyric-acid)-releasing neurons of the lateral hypothalamus, which promote the transition to wakefulness from non-rapid eye movement (NREM) and rapid eye movement (REM) sleep2,3. Here we show that a subset of GABAergic neurons in the mouse ventral zona incerta, which express the LIM homeodomain factor Lhx6 and are activated by sleep pressure, both directly inhibit wake-active hypocretin and GABAergic cells in the lateral hypothalamus and receive inputs from multiple sleep–wake-regulating neurons. Conditional deletion of Lhx6 from the developing diencephalon leads to decreases in both NREM and REM sleep. Furthermore, selective activation and inhibition of Lhx6-positive neurons in the ventral zona incerta bidirectionally regulate sleep time in adult mice, in part through hypocretin-dependent mechanisms. These studies identify a GABAergic subpopulation of neurons in the ventral zona incerta that promote sleep.

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Acknowledgements

We thank L. de Lecea and A. Gittis for providing mice; M. Pletnikov for help with behavioural analysis; and M. Wu, V. Mongrain, R. Kuruvila, D. Lee, J. Bedont and W. Yap for comments on the manuscript. This work was supported by a Johns Hopkins Discovery Fund award to S.B. and S.H. S.P.B. is supported by a Klingenstein-Simons Foundation Fellowship in the Neurosciences.

Author information

    • Juhyun Kim
    • , Dong Won Kim
    •  & Yi Stephanie Zhang

    These authors contributed equally to this work.

Affiliations

  1. Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    • Kai Liu
    • , Juhyun Kim
    • , Dong Won Kim
    • , Yi Stephanie Zhang
    • , Chang Liu
    • , Solange P. Brown
    •  & Seth Blackshaw

  2. Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA

    • Hechen Bao
    • , Szu-Aun Lim
    •  & Juan Song

  3. The Francis Crick Institute, London, UK

    • Myrto Denaxa
    •  & Vassilis Pachnis

  4. Department of Biology, Johns Hopkins University, Baltimore, Maryland, USA

    • Eileen Kim

  5. The McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    • Ian R. Wickersham

  6. National Institute of Mental Health, Bethesda, Maryland, USA

    • Samer Hattar

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Contributions

K.L. and S.B. designed the study. K.L., D.W.K. and Y.S.Z. conducted immunostaining. C.L. conducted statistical analysis. J.K. and S.P.B. designed, conducted and analysed slice electrophysiology experiments. H.B., S.-A.L and J.S. performed rabies virus experiments. I.R.W. provided rabies virus. M.D. and V.P. generated Lhx6lox/lox mice. K.L. performed all other surgeries and behavioural analysis, with assistance from D.W.K. and E.K., under the guidance of S.H. K.L., D.W.K., J.K., S.H., S.P.B. and S.B. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Seth Blackshaw.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended data figures

  1. 1.

    Distribution of diencephalic Lhx6+ neurons and immunohistochemical analysis of Lhx6-expressing neurons.

  2. 2.

    Lhx6-Cre activity recapitulates endogenous Lhx6 expression pattern and immunohistochemical analysis of Lhx6-expressing neurons.

  3. 3.

    Additional data showing mapping of neural projection sites of Lhx6-expressing neurons in the VZI, and projection sites of Slc32a1 and Pvalb-expressing cells in the zona incerta.

  4. 4.

    Presynaptic inputs to Lhx6+ zona incerta neurons.

  5. 5.

    Synaptic outputs of Lhx6 neurons in the zona incerta.

  6. 6.

    Lhx6 expression is selectively deleted in the diencephalon, but preserved in the telencephalon of Lhx6-conditional knockout Foxd1-cre;Lhx6lox/lox (cKO) line, and Lhx6 cKO mice do not display obvious behavioural abnormalities other than changes in sleep patterns.

  7. 7.

    Distribution of AAV-DREADD infection sites in the zona incerta of Lhx6-cre mice and CNO activates Fos expression in AAV-DIO-hM3Dq-injected Lhx6-Cre mice, whereas CNO does not affect sleep in AAV-DIO–mCherry-injected Lhx6-Cre mice.

  8. 8.

    Additional data from Gq DREADD-injected Lhx6-cre mice.

  9. 9.

    The dual orexin receptor antagonist suvorexant can increase both NREM and REM sleep.

  10. 10.

    Additional data from AAV-DIO-hM4Di-injected Lhx6-cre mice.

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DOI

https://doi.org/10.1038/nature23663

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