Brain Structure and Function

, Volume 224, Issue 1, pp 99–110 | Cite as

Melanin-concentrating hormone neurons promote rapid eye movement sleep independent of glutamate release

  • Fumito Naganuma
  • Sathyajit S. Bandaru
  • Gianna Absi
  • Melissa J. Chee
  • Ramalingam VetrivelanEmail author
Original Article


Neurons containing melanin-concentrating hormone (MCH) in the posterior lateral hypothalamus play an integral role in rapid eye movement sleep (REMs) regulation. As MCH neurons also contain a variety of other neuropeptides [e.g., cocaine- and amphetamine-regulated transcript (CART) and nesfatin-1] and neurotransmitters (e.g., glutamate), the specific neurotransmitter responsible for REMs regulation is not known. We hypothesized that glutamate, the primary fast-acting neurotransmitter in MCH neurons, is necessary for REMs regulation. To test this hypothesis, we deleted vesicular glutamate transporter (Vglut2; necessary for synaptic release of glutamate) specifically from MCH neurons by crossing MCH-Cre mice (expressing Cre recombinase in MCH neurons) with Vglut2flox/flox mice (expressing LoxP-modified alleles of Vglut2), and studied the amounts, architecture and diurnal variation of sleep-wake states during baseline conditions. We then activated the MCH neurons lacking glutamate neurotransmission using chemogenetic methods and tested whether these MCH neurons still promoted REMs. Our results indicate that glutamate in MCH neurons contributes to normal diurnal variability of REMs by regulating the levels of REMs during the dark period, but MCH neurons can promote REMs even in the absence of glutamate.


Paradoxical sleep Lateral hypothalamus Conditional knockout Chemogenetics Locomotor activity Body temperature Diurnal rhythms 



We thank Quan Ha, Minh Ha and Celia Gagliardi for excellent technical assistance and Dr. Daniel Kroeger for his help with histology imaging. We also thank Dr. Eleftheria Maratos-Flier (Department of Medicine, Beth Israel Deaconess Medical Center, Boston) for providing the rabbit anti-MCH antibody.

Author contributions

Conceived and designed the experiments: FN and RV; Performed the experiments: FN, SB, GA and RV; Analyzed the data: FN, SB, GA and RV; Written the manuscript: FN, SB and RV with input from all authors. Provided key reagents: MC; Supervision: RV.


This work was supported by National Institutes of Health Grants [R21-NS074205, R01-NS088482 (to RV)].

Compliance with ethical standards

Conflict of interest

This was not an industry-supported study. The authors declare that they have no competing interests.

Research involving human participants and/or animals

Care of the animals met National Institutes of Health standards, as set forth in the Guide for the Care and Use of Laboratory Animals, and all protocols were approved by the BIDMC Institutional Animal Care and Use Committee.


  1. Adamantidis A et al (2008) Sleep architecture of the melanin-concentrating hormone receptor 1-knockout mice. Eur J Neurosci 27:1793–1800. CrossRefGoogle Scholar
  2. Ahnaou A, Dautzenberg FM, Huysmans H, Steckler T, Drinkenburg WH (2011) Contribution of melanin-concentrating hormone (MCH1) receptor to thermoregulation and sleep stabilization: evidence from MCH1(−/−) mice. Behav Brain Res 218:42–50. CrossRefGoogle Scholar
  3. Benedetto L, Rodriguez-Servetti Z, Lagos P, D’Almeida V, Monti JM, Torterolo P (2013) Microinjection of melanin concentrating hormone into the lateral preoptic area promotes non-REM sleep in the rat. Peptides 39:11–15 CrossRefGoogle Scholar
  4. Bittencourt JC et al (1992) The melanin-concentrating hormone system of the rat brain: an immuno- and hybridization histochemical characterization. J Comp Neurol 319:218–245. CrossRefGoogle Scholar
  5. Brischoux F, Cvetkovic V, Griffond B, Fellmann D, Risold PY (2002) Time of genesis determines projection and neurokinin-3 expression patterns of diencephalic neurons containing melanin-concentrating hormone. Eur J Neurosci 16:1672–1680CrossRefGoogle Scholar
  6. Chee MJ, Arrigoni E, Maratos-Flier E (2015a) Melanin-concentrating hormone neurons release glutamate for feedforward inhibition of the lateral septum. J Neurosci 35:3644–3651. CrossRefGoogle Scholar
  7. Chee MJ et al (2015b) Melanin-concentrating hormone is necessary for olanzapine-inhibited locomotor activity in male mice. Eur Neuropsychopharmacol 25:1808–1816. CrossRefGoogle Scholar
  8. Chou TC, Scammell TE, Gooley JJ, Gaus SE, Saper CB, Lu J (2003) Critical role of dorsomedial hypothalamic nucleus in a wide range of behavioral circadian rhythms. J Neurosci 23:10691–10702CrossRefGoogle Scholar
  9. Cvetkovic V, Brischoux F, Griffond B, Bernard G, Jacquemard C, Fellmann D, Risold PY (2003a) Evidence of melanin-concentrating hormone-containing neurons supplying both cortical neuroendocrine projections. Neuroscience 116:31–35CrossRefGoogle Scholar
  10. Cvetkovic V, Poncet F, Fellmann D, Griffond B, Risold PY (2003b) Diencephalic neurons producing melanin-concentrating hormone are influenced by local and multiple extra-hypothalamic tachykininergic projections through the neurokinin 3 receptor. Neuroscience 119:1113–1145CrossRefGoogle Scholar
  11. Cvetkovic V, Brischoux F, Jacquemard C, Fellmann D, Griffond B, Risold PY (2004) Characterization of subpopulations of neurons producing melanin-concentrating hormone in the rat ventral diencephalon. J Neurochem 91:911–919. CrossRefGoogle Scholar
  12. Ferreira JGP, Bittencourt JC, Adamantidis A (2017) Melanin-concentrating hormone and sleep. Curr Opin Neurobiol 44:152–158. CrossRefGoogle Scholar
  13. Fujimoto M, Fukuda S, Sakamoto H, Takata J, Sawamura S (2017) Neuropeptide glutamic acid-isoleucine (NEI)-induced paradoxical sleep in rats. Peptides 87:28–33. CrossRefGoogle Scholar
  14. Glick M, Segal-Lieberman G, Cohen R, Kronfeld-Schor N (2009) Chronic MCH infusion causes a decrease in energy expenditure and body temperature, and an increase in serum IGF-1 levels in mice. Endocrine 36:479–485. CrossRefGoogle Scholar
  15. Hanriot L, Camargo N, Courau AC, Leger L, Luppi PH, Peyron C (2007) Characterization of the melanin-concentrating hormone neurons activated during paradoxical sleep hypersomnia in rats. J Comp Neurol 505:147–157. CrossRefGoogle Scholar
  16. Hassani OK, Lee MG, Jones BE (2009) Melanin-concentrating hormone neurons discharge in a reciprocal manner to orexin neurons across the sleep-wake cycle. Proc Natl Acad Sci USA 106:2418–2422. CrossRefGoogle Scholar
  17. Jego S, Salvert D, Renouard L, Mori M, Goutagny R, Luppi PH, Fort P (2012) Tuberal hypothalamic neurons secreting the satiety molecule Nesfatin-1 are critically involved in paradoxical (REM) sleep homeostasis. PLoS One 7:e52525. CrossRefGoogle Scholar
  18. Jego S et al (2013) Optogenetic identification of a rapid eye movement sleep modulatory circuit in the hypothalamus. Nat Neurosci 16:1637–1643. CrossRefGoogle Scholar
  19. Keating GL, Kuhar MJ, Bliwise DL, Rye DB (2010) Wake promoting effects of cocaine and amphetamine-regulated transcript (CART). Neuropeptides 44:241–246. CrossRefGoogle Scholar
  20. Kitka T et al (2011) Association between the activation of MCH and orexin immunorective neurons and REM sleep architecture during REM rebound after a three day long REM deprivation. Neurochem Int 59:686–694. CrossRefGoogle Scholar
  21. Konadhode RR et al (2013) Optogenetic stimulation of MCH neurons increases sleep. J Neurosci 33:10257–10263. CrossRefGoogle Scholar
  22. Kong D et al (2010) Glucose stimulation of hypothalamic MCH neurons involves K(ATP) channels, is modulated by UCP2, and regulates peripheral glucose homeostasis. Cell Metab 12:545–552. CrossRefGoogle Scholar
  23. Krenzer M et al (2011) Brainstem and spinal cord circuitry regulating REM sleep and muscle atonia. PLoS One 6:e24998. CrossRefGoogle Scholar
  24. Lu J, Greco MA, Shiromani P, Saper CB (2000) Effect of lesions of the ventrolateral preoptic nucleus on NREM and REM sleep. J Neurosci 20:3830–3842CrossRefGoogle Scholar
  25. Mickelsen LE et al (2017) Neurochemical heterogeneity among lateral hypothalamic hypocretin/orexin and melanin-concentrating hormone neurons identified through single-cell gene expression analysis. eNeuro. Google Scholar
  26. Monti JM, Torterolo P, Lagos P (2013) Melanin-concentrating hormone control of sleep-wake behavior. Sleep Med Rev 17:293–298. CrossRefGoogle Scholar
  27. Sapin E, Berod A, Leger L, Herman PA, Luppi PH, Peyron C (2010) A very large number of GABAergic neurons are activated in the tuberal hypothalamus during paradoxical (REM) sleep hypersomnia. PLoS One 5:e11766 CrossRefGoogle Scholar
  28. Scammell T, Gerashchenko D, Urade Y, Onoe H, Saper C, Hayaishi O (1998) Activation of ventrolateral preoptic neurons by the somnogen prostaglandin D2. Proc Natl Acad Sci USA 95:7754–7759CrossRefGoogle Scholar
  29. Schneeberger M et al (2018) Functional analysis reveals differential effects of glutamate and MCH neuropeptide in MCH neurons. Mol Metab 13:83–89. CrossRefGoogle Scholar
  30. Tong Q et al (2007) Synaptic glutamate release by ventromedial hypothalamic neurons is part of the neurocircuitry that prevents hypoglycemia. Cell Metab 5:383–393. CrossRefGoogle Scholar
  31. Torterolo P, Lagos P, Monti JM (2011) Melanin-concentrating hormone: a new sleep factor? Front Neurol 2:14. CrossRefGoogle Scholar
  32. Tsunematsu T et al (2014) Optogenetic manipulation of activity and temporally controlled cell-specific ablation reveal a role for MCH neurons in sleep/wake regulation. J Neurosci 34:6896–6909. CrossRefGoogle Scholar
  33. Varin C, Luppi PH, Fort P (2018) Melanin-concentrating hormone-expressing neurons adjust slow-wave sleep dynamics to catalyze paradoxical (REM) sleep. Sleep. Google Scholar
  34. Vas S et al (2013) Nesfatin-1/NUCB2 as a potential new element of sleep regulation in rats. PLoS One 8:e59809. CrossRefGoogle Scholar
  35. Verret L et al (2003) A role of melanin-concentrating hormone producing neurons in the central regulation of paradoxical sleep. BMC Neurosci 4:19. CrossRefGoogle Scholar
  36. Vetrivelan R, Fuller PM, Tong Q, Lu J (2009) Medullary circuitry regulating rapid eye movement sleep and motor atonia. J Neurosci 29:9361–9369. CrossRefGoogle Scholar
  37. Vetrivelan R et al (2016) Melanin-concentrating hormone neurons specifically promote rapid eye movement sleep in mice. Neuroscience 336:102–113. CrossRefGoogle Scholar
  38. Whiddon BB, Palmiter RD (2013) Ablation of neurons expressing melanin-concentrating hormone (MCH) in adult mice improves glucose tolerance independent of MCH signaling. J Neurosci 33:2009–2016. CrossRefGoogle Scholar
  39. Willie JT, Sinton CM, Maratos-Flier E, Yanagisawa M (2008) Abnormal response of melanin-concentrating hormone deficient mice to fasting: hyperactivity and rapid eye movement sleep suppression. Neuroscience 156:819–829. CrossRefGoogle Scholar
  40. Yamashita T, Yamanaka A (2017) Lateral hypothalamic circuits for sleep-wake control. Curr Opin Neurobiol 44:94–100. CrossRefGoogle Scholar
  41. Zhou D, Shen Z, Strack AM, Marsh DJ, Shearman LP (2005) Enhanced running wheel activity of both Mch1r- and Pmch-deficient mice. Regul Pept 124:53–63. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Neurology, Beth Israel Deaconess Medical Center and Division of Sleep MedicineHarvard Medical SchoolBostonUSA
  2. 2.Division of Pharmacology, Faculty of MedicineTohoku Medical and Pharmaceutical UniversitySendaiJapan
  3. 3.Department of NeuroscienceCarleton UniversityOttawaCanada

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