Diverse roles of pontine NPS-expressing neurons in sleep regulation

Contributed by Ying-Hui Fu; received November 17, 2023; accepted January 17, 2024; reviewed by Paul Franken and Yu-Ting Tseng
February 21, 2024
121 (9) e2320276121

Significance

We previously identified a mutation in the only receptor for the neuropeptide S (NPS), NPSR1 (NPS receptor 1), that enables people and mice to have reduced sleep duration. NPS is an important neurotransmitter that has been shown to play roles in regulating arousal, mood, food intake, and memory. Since the neuro-network involved in sleep/wake behavior is expected to be complex, we set out to investigate the role of NPS in various brain nuclei as a step toward further mapping the network. We found that NPS neurons in three different nuclei are either wake-promoting, sleep-promoting, or without effect, highlighting the intricate complexity of the sleep/wake-modulating network.

Abstract

Neuropeptide S (NPS) was postulated to be a wake-promoting neuropeptide with unknown mechanism, and a mutation in its receptor (NPSR1) causes the short sleep duration trait in humans. We investigated the role of different NPS+ nuclei in sleep/wake regulation. Loss-of-function and chemogenetic studies revealed that NPS+ neurons in the parabrachial nucleus (PB) are wake-promoting, whereas peri-locus coeruleus (peri-LC) NPS+ neurons are not important for sleep/wake modulation. Further, we found that a NPS+ nucleus in the central gray of the pons (CGPn) strongly promotes sleep. Fiber photometry recordings showed that NPS+ neurons are wake-active in the CGPn and wake/REM-sleep active in the PB and peri-LC. Blocking NPS–NPSR1 signaling or knockdown of Nps supported the function of the NPS–NPSR1 pathway in sleep/wake regulation. Together, these results reveal that NPS and NPS+ neurons play dichotomous roles in sleep/wake regulation at both the molecular and circuit levels.

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Data, Materials, and Software Availability

All study data are included in the article and/or SI Appendix.

Acknowledgments

We thank Dr. Patrick Hsu at UC Berkeley for the CasRx plasmid and all other members in the Ptáček and Fu labs for helpful discussions. This work was supported by NIH grants NS117929 to L.J.P. and NS072360 and NS104782 to Y.-H.F. The generation of mouse models was also supported by NIH P30 DK063720 to the Diabetes Center at University of California San Francisco.

Author contributions

L.X., L.J.P., and Y.-H.F. designed research; L.X., X.Z., C.Y., J.M.W., and G.S. performed research; L.X., X.Z., C.Y., G.S., and Y.-H.F. analyzed data; and L.X., X.Z., G.S., L.J.P., and Y.-H.F. wrote the paper.

Competing interests

The authors declare no competing interest.

Supporting Information

Appendix 01 (PDF)

References

1
T. E. Scammell, E. Arrigoni, J. O. Lipton, Neural circuitry of wakefulness and sleep. Neuron 93, 747–765 (2017), https://doi.org/10.1016/j.neuron.2017.01.014.
2
D. Liu, Y. Dan, A motor theory of sleep-wake control: Arousal-action circuit. Annu. Rev. Neurosci. 42, 27–46 (2019), https://doi.org/10.1146/annurev-neuro-080317-061813.
3
F. Weber, Y. Dan, Circuit-based interrogation of sleep control. Nature 538, 51–59 (2016).
4
F. Tatsuki et al., Involvement of Ca2+-dependent hyperpolarization in sleep duration in mammals. Neuron 90, 70–85 (2016).
5
Y. Niwa et al., Muscarinic acetylcholine receptors Chrm1 and Chrm3 are essential for REM sleep. Cell Rep. 24, 2231–2247.e7 (2018), https://doi.org/10.1016/j.celrep.2018.07.082.
6
L. Lin et al., The sleep disorder canine narcolepsy is caused by a mutation in the hypocretin (orexin) receptor 2 gene. Cell 98, 365–376 (1999), https://doi.org/10.1016/S0092-8674(00)81965-0.
7
K. Yoshida et al., Leak potassium channels regulate sleep duration. Proc. Natl. Acad. Sci. U.S.A. 115, E9459–E9468 (2018), https://doi.org/10.1073/pnas.1806486115.
8
H. Funato et al., Forward-genetics analysis of sleep in randomly mutagenized mice. Nature 539, 378–383 (2016).
9
G. Shi et al., A rare mutation of β1-adrenergic receptor affects sleep/wake behaviors. Neuron 103, 1044–1055.e7 (2019), https://doi.org/10.1016/j.neuron.2019.07.026.
10
L. Xing et al., Mutant neuropeptide S receptor reduces sleep duration with preserved memory consolidation. Sci. Transl. Med. 11, eaax2014 (2019), https://doi.org/10.1126/scitranslmed.aax2014.
11
G. Shi et al., Mutations in metabotropic glutamate receptor 1 contribute to natural short sleep trait. Curr. Biol. 31, 13–24.e4 (2020), https://doi.org/10.1016/j.cub.2020.09.071.
12
G. A. Sunagawa et al., Mammalian reverse genetics without crossing reveals Nr3a as a short-sleeper gene. Cell Rep. 14, 662–677 (2016), https://doi.org/10.1016/j.celrep.2015.12.052.
13
K. Koh et al., Identification of SLEEPLESS, a sleep-promoting factor. Science 321, 372–376 (2008).
14
C. Cirelli et al., Reduced sleep in Drosophila Shaker mutants. Nature 434, 1087–1092 (2005).
15
P. Franken et al., NPAS2 as a transcriptional regulator of non-rapid eye movement sleep: Genotype and sex interactions. Proc. Natl. Acad. Sci. U.S.A. 103, 7118–7123 (2006).
16
A. Vassalli, P. Franken, Hypocretin (orexin) is critical in sustaining theta/gamma-rich waking behaviors that drive sleep need. Proc. Natl. Acad. Sci. U.S.A. 114, E5464–E5473 (2017).
17
P. Franken, D. Chollet, M. Tafti, The homeostatic regulation of sleep need is under genetic control. J. Neurosci. 21, 2610–2621 (2001).
18
R. K. Reinscheid, Y. L. Xu, Neuropeptide S as a novel arousal promoting peptide transmitter. FEBS J. 272, 5689–5693 (2005), https://doi.org/10.1111/j.1742-4658.2005.04982.x.
19
Y. L. Xu et al., Neuropeptide S: A neuropeptide promoting arousal and anxiolytic-like effects. Neuron 43, 487–497 (2004), https://doi.org/10.1016/j.neuron.2004.08.005.
20
M. Lukas, I. D. Neumann, Nasal application of neuropeptide S reduces anxiety and prolongs memory in rats: Social versus non-social effects. Neuropharmacology 62, 398–405 (2012), https://doi.org/10.1016/j.neuropharm.2011.08.016.
21
K. L. Smith et al., Neuropeptide S stimulates the hypothalamo-pituitary-adrenal axis and inhibits food intake. Endocrinology 147, 3510–3518 (2006), https://doi.org/10.1210/en.2005-1280.
22
Y. L. Peng et al., Central neuropeptide S inhibits food intake in mice through activation of neuropeptide S receptor. Peptides 31, 2259–2263 (2010), https://doi.org/10.1016/j.peptides.2010.08.015.
23
M. Niimi, Centrally administered neuropeptide S activates orexin-containing neurons in the hypothalamus and stimulates feeding in rats. Endocrine 30, 75–79 (2006), https://doi.org/10.1385/ENDO:30:1:75.
24
J. Cao, L. De Lecea, S. Ikemoto, Intraventricular administration of neuropeptide S has reward-like effects. Eur. J. Pharmacol. 658, 16–21 (2011), https://doi.org/10.1016/j.ejphar.2011.02.009.
25
A. Pulga, C. Ruzza, A. Rizzi, R. Guerrini, G. Calo, Anxiolytic- and panicolytic-like effects of neuropeptide S in the mouse elevated T-maze. Eur. J. Neurosci. 36, 3531–3537 (2012), https://doi.org/10.1111/j.1460-9568.2012.08265.x.
26
G. Wegener et al., Neuropeptide S alters anxiety, but not depression-like behaviour in Flinders Sensitive Line rats: A genetic animal model of depression. Int. J. Neuropsychopharmacol. 15, 375–387 (2012), https://doi.org/10.1017/S1461145711000678.
27
K. Jüngling et al., Neuropeptide S-mediated control of fear expression and extinction: Role of intercalated GABAergic neurons in the amygdala. Neuron 59, 298–310 (2008), https://doi.org/10.1016/j.neuron.2008.07.002.
28
N. Okamura et al., Neuropeptide S enhances memory during the consolidation phase and interacts with noradrenergic systems in the brain. Neuropsychopharmacology 36, 744–752 (2011), https://doi.org/10.1038/npp.2010.207.
29
C. Ruzza et al., Behavioural phenotypic characterization of CD-1 mice lacking the neuropeptide S receptor. Neuropharmacology 62, 1999–2009 (2012), https://doi.org/10.1016/j.neuropharm.2011.12.036.
30
S. D. Clark et al., Anatomical characterization of the neuropeptide S system in the mouse brain by in situ hybridization and immunohistochemistry. J. Comp. Neurol. 519, 1867–1893 (2011), https://doi.org/10.1002/cne.22606.
31
X. Liu et al., Molecular fingerprint of neuropeptide s-producing neurons in the mouse brain. J. Comp. Neurol. 519, 1847–1866 (2011), https://doi.org/10.1002/cne.22603.
32
D. Huang et al., Neuropeptide S (NPS) neurons: Parabrachial identity and novel distributions. J. Comp. Neurol. 530, 3157–3178 (2022).
33
J. Liu et al., Cell-specific translational profiling in acute kidney injury. J. Clin. Invest. 124, 1242–1254 (2014), https://doi.org/10.1172/JCI72126.
34
S. Han, M. Soleiman, M. Soden, L. Zweifel, R. D. Palmiter, Elucidating an affective pain circuit that creates a threat memory. Cell 162, 363–374 (2015).
35
J. C. Kim et al., Linking genetically defined neurons to behavior through a broadly applicable silencing allele. Neuron 63, 305–315 (2009), https://doi.org/10.1016/j.neuron.2009.07.010.
36
G. M. Alexander et al., Remote control of neuronal activity in transgenic mice expressing evolved G protein-coupled receptors. Neuron 63, 27–39 (2009).
37
C. Ruzza et al., Further studies on the pharmacological profile of the neuropeptide S receptor antagonist SHA 68. Peptides 31, 915–925 (2010), https://doi.org/10.1016/j.peptides.2010.02.012.
38
N. Okamura, S. A. Habay, J. Zeng, A. R. Chamberlin, R. K. Reinscheid, Synthesis and pharmacological in vitro and in vivo profile of 3-oxo-1,1-diphenyl-tetrahydro-oxazolo[3,4-a]pyrazine-7-carboxylic acid 4-fluoro-benzylamide (SHA 68), a selective antagonist of the neuropeptide S receptor. J. Pharmacol. Exp. Ther. 325, 893–901 (2008), https://doi.org/10.1124/jpet.107.135103.
39
C. Adori et al., Neuropeptide S-and neuropeptide S receptor-expressing neuron populations in the human pons. Front. Neuroanat. 9, 126 (2015), https://doi.org/10.3389/fnana.2015.00126.
40
N. R. Wall, I. R. Wickersham, A. Cetin, M. De La Parra, E. M. Callaway, Monosynaptic circuit tracing in vivo through Cre-dependent targeting and complementation of modified rabies virus. Proc. Natl. Acad. Sci. U.S.A. 107, 21848–21853 (2010), https://doi.org/10.1073/pnas.1011756107.
41
I. R. Wickersham et al., Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons. Neuron 53, 639–647 (2007), https://doi.org/10.1016/j.neuron.2007.01.033.
42
T. K. Lavin, L. Jin, N. E. Lea, I. R. Wickersham, Monosynaptic tracing success depends critically on helper virus concentrations. Front. Synaptic Neurosci. 12, 6 (2020), https://doi.org/10.3389/fnsyn.2020.00006.
43
M. Xu et al., Basal forebrain circuit for sleep-wake control. Nat. Neurosci. 18, 1641–1647 (2015).
44
R. R. Konadhode, D. Pelluru, P. J. Shiromani, Neurons containing orexin or melanin concentrating hormone reciprocally regulate wake and sleep. Front. Syst. Neurosci. 8, 1–9 (2015).
45
H. Qin et al., REM sleep-active hypothalamic neurons may contribute to hippocampal social-memory consolidation. Neuron 110, 4000–4014.e6 (2022).
46
S. Izawa et al., REM sleep-active MCH neurons are involved in forgetting hippocampusdependent memories. Science 365, 1308–1313 (2019).
47
P. Franken, D.-J. Dijk, Sleep and circadian rhythmicity as entangled processes serving homeostasis. Nat. Rev. Neurosci. 25, 43–59 (2023), https://doi.org/10.1038/s41583-023-00764-z.
48
Y. Q. Wang, W. Y. Liu, L. Li, W. M. Qu, Z. L. Huang, Neural circuitry underlying REM sleep: A review of the literature and current concepts. Prog. Neurobiol. 204, 102106 (2021).

Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 121 | No. 9
February 27, 2024
PubMed: 38381789

Classifications

Data, Materials, and Software Availability

All study data are included in the article and/or SI Appendix.

Submission history

Received: November 17, 2023
Accepted: January 17, 2024
Published online: February 21, 2024
Published in issue: February 27, 2024

Keywords

  1. NPS
  2. parabrachial nucleus
  3. central gray of the pons
  4. sleep
  5. neural circuitry

Acknowledgments

We thank Dr. Patrick Hsu at UC Berkeley for the CasRx plasmid and all other members in the Ptáček and Fu labs for helpful discussions. This work was supported by NIH grants NS117929 to L.J.P. and NS072360 and NS104782 to Y.-H.F. The generation of mouse models was also supported by NIH P30 DK063720 to the Diabetes Center at University of California San Francisco.
Author Contributions
L.X., L.J.P., and Y.-H.F. designed research; L.X., X.Z., C.Y., J.M.W., and G.S. performed research; L.X., X.Z., C.Y., G.S., and Y.-H.F. analyzed data; and L.X., X.Z., G.S., L.J.P., and Y.-H.F. wrote the paper.
Competing Interests
The authors declare no competing interest.

Notes

Reviewers: P.F., Universite de Lausanne; and Y.-T.T., Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences.

Authors

Affiliations

Lijuan Xing
Department of Neurology, University of California San Francisco, San Francisco, CA 94143
Xianlin Zou
Department of Neurology, University of California San Francisco, San Francisco, CA 94143
Chen Yin
Department of Neurology, University of California San Francisco, San Francisco, CA 94143
John M. Webb
Department of Neurology, University of California San Francisco, San Francisco, CA 94143
Guangsen Shi
Zhongshan Institute for Drug Discovery, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Zhongshan 528400, China
Louis J. Ptáček1 [email protected]
Department of Neurology, University of California San Francisco, San Francisco, CA 94143
Department of Neurology, Weill Institute for Neurosciences, University of California San Francisco, San Francisco, CA 94143
Kavli Institute for Fundamental Neuroscience, University of California San Francisco, San Francisco, CA 94143
Institute of Human Genetics, University of California San Francisco, San Francisco, CA 94143
Department of Neurology, University of California San Francisco, San Francisco, CA 94143
Department of Neurology, Weill Institute for Neurosciences, University of California San Francisco, San Francisco, CA 94143
Kavli Institute for Fundamental Neuroscience, University of California San Francisco, San Francisco, CA 94143
Institute of Human Genetics, University of California San Francisco, San Francisco, CA 94143

Notes

1
To whom correspondence may be addressed. Email: [email protected] or [email protected].

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Diverse roles of pontine NPS-expressing neurons in sleep regulation
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