Orexins (also called hypocretins), which are neuropeptides exclusively expressed by a population of neurons specifically localized in the lateral hypothalamic area, are critically implicated in the regulation of sleep/wake states. Orexin deficiency results in narcoleptic phenotype in rodents, dogs, and humans, suggesting that orexins are important for maintaining consolidated wakefulness states. However, the physiological effect of constitutive increased orexinergic transmission tone, which might be important for understanding the effects of orexin agonists that are promising candidates for therapeutic agents of narcolepsy, has not been fully characterized. We report here the sleep/wakefulness abnormalities in transgenic mice that exhibit widespread overexpression of a rat prepro-orexin transgene driven by a β-actin/cytomegalovirus hybrid promoter (CAG/orexin transgenic mice). CAG/orexin mice exhibit sleep abnormalities with fragmentation of non-rapid eye movement (REM) sleep episode and a reduction in REM sleep. Non-REM sleep was frequently disturbed by short episodes of wakefulness. EEG/EMG studies also reveal incomplete REM sleep atonia with abnormal myoclonic activity during this sleep stage. These results suggest that endogenous orexinergic activity should be appropriately regulated for normal maintenance of sleep states. Orexinergic transmission should be activated during wakefulness, while it should be inactivated or decreased during sleep state to maintain appropriate vigilance states.

Orexin is a neuropeptide that plays a highly important role in mechanisms that regulate sleep/wake states. Lack of the orexin gene or orexin-producing neurons (orexin neurons) results in narcolepsy in several mammalian species, suggesting that orexin is an important factor for the maintenance of wakefulness. Constitutive, ectopic expression of orexin in transgenic mice resulted in severe fragmentation of non–rapid eye movement sleep, along with abnormal muscle tone regulation during REM sleep, suggesting that activity of orexin neurons should be appropriately decreased during sleep to maintain consolidated sleep states. This review will discuss the mechanisms by which the orexin system is regulated during sleep.

The neuropeptides orexin A and orexin B (also known as hypocretin 1 and hypocretin 2), produced in lateral hypothalamic neurons, are critical regulators of feeding behavior, the reward system, and sleep/wake states. Orexin-producing neurons (orexin neurons) are regulated by various factors involved in regulation of energy homeostasis and sleep/wakefulness states. Bombesin receptor subtype 3 (BRS3) is an orphan receptor that might be implicated in energy homeostasis and is highly expressed in the hypothalamus. However, the neural pathway by which BRS3 regulates energy homeostasis is largely unknown. We examined whether BRS3 is involved in the regulation of orexin neurons. Using a calcium imaging method, we found that a selective BRS3 agonist [Ac-Phe-Trp-Ala-His-(τBzl)-Nip-Gly-Arg-NH2] increased the intracellular calcium concentration of orexin neurons. However, intracellular recordings from slice preparations revealed that the BRS3 agonist hyperpolarized orexin neurons. The BRS3 agonist depolarized orexin neuron in the presence of tetrodotoxin. Moreover, in the presence of GABA receptor blockers, picrotoxin and CGP55845, the BRS3 agonist induced depolarization and increased firing frequency. Additionally, double-label in situ hybridization study revealed that Brs3 mRNA was expressed in almost all orexin neurons and many cells around these neurons. These findings suggest that the BRS3 agonist indirectly inhibited orexin neurons through GABAergic input and directly activated orexin neurons. Inhibition of activity of orexin neurons through BRS3 might be an important pathway for regulation of feeding and sleep/wake states. This pathway might serve as a novel target for the treatment of obesity.

GPR109B (HM74) is a putative G protein-coupled receptor (GPCR) whose cognate ligands have yet to be characterized. GPR109B shows a high degree of sequence similarity to GPR109A, another GPCR that was identified as a high-affinity nicotinic acid (niacin) receptor. However, the affinity of nicotinic acid to GPR109B is very low. In this study, we found that certain aromatic D-amino acids, including D-phenylalanine, D-tryptophan, and the metabolite of the latter, D-kynurenine, decreased the activity of adenylate cyclase in cells transfected with GPR109B cDNA through activation of pertussis toxin (PTX)-sensitive G proteins. These D-amino acids also elicited a transient rise of intracellular Ca2+ level in cells expressing GPR109B in a PTX-sensitive manner. In contrast, these D-amino acids did not show any effects on cells expressing GPR109A. We found that the GPR109B mRNA is abundantly expressed in human neutrophils. D-phenylalanine and D-tryptophan induced a transient increase of intracellular Ca2+ level and a reduction of cAMP levels in human neutrophils. Furthermore, knockdown of GPR109B by RNA interference inhibited the D-amino acids-induced decrease of cellular cAMP levels in human neutrophils. These D-amino acids induced chemotactic activity of freshly prepared human neutrophils. We also found that D-phenylalanine and D-tryptophan induced chemotactic responses in Jurkat cells transfected with the GPR109B cDNA but not in mock-transfected Jurkat cells. These results suggest that these aromatic D-amino acids elicit a chemotactic response in human neutrophils via activation of GPR109B.

Hypothalamic neurons that contain the neuropeptide orexin (hypocretin) play important roles in the regulation of sleep/wake. Here we analyze the in vivo and in vitro phenotype of mice lacking the GABAB1 gene specifically in orexin neurons (oxGKO mice) and demonstrate that GABAB receptors on orexin neurons are essential in stabilizing and consolidating sleep/wake states. In oxGKO brain slices, we show that the absence of GABAB receptors decreases the sensitivity of orexin neurons to both excitatory and inhibitory inputs because of augmented GABAA-mediated inhibition that increases the membrane conductance and shunts postsynaptic currents in these neurons. This increase in GABAA-mediated inhibitory tone is apparently the result of an orexin receptor type 1-mediated activation of local GABAergic interneurons that project back onto orexin neurons. oxGKO mice exhibit severe fragmentation of sleep/wake states during both the light and dark periods, without showing an abnormality in total sleep time or signs of cataplexy. Thus, GABAB receptors on orexin neurons are crucial in the appropriate control of the orexinergic tone through sleep/wake states, thereby stabilizing the state switching mechanisms.

Lateral hypothalamic neuropeptides, orexins, have been recognized as one of the most important regulators of sleep/wakefulness states. Besides, these peptides are also regarded as an important factor that regulates feeding behavior, owing to their localization within the lateral hypothalamic area, the classic “feeding center”, pharmacological activities, and the fact that prepro-orexin mRNA is upregulated when animals are fasted. This review summarizes the role of orexins in the regulation of feeding behavior and body weight homeostasis in relation to other systems that involve orexinergic neurotransmission.

Avoiding danger and finding food, which are life-sustaining activities that are regulated by emotion, reward and energy balance, require proper wakefulness. The orexin system controls sleep and wakefulness through interactions with systems that regulate emotion, reward and energy homeostasis. Recent findings have brought about the possibility of novel therapies targeting the orexin system for sleep disorders, including insomnia and narcolepsy–cataplexy, as well as other pathological conditions such as obesity and drug addiction [1]. In this review, we will discuss the current understanding of the integrative physiology and clinical perspectives of the orexin system. We will briefly review signaling through orexin A and B receptors and discuss the role of orexins in the pathophysiology of narcolepsy. We will also examine connections between orexin neurons and other brain areas involved in feeding behavior, reward and emotion. Finally, we will consider the therapeutic potential of drugs that target orexin receptors.

Decrease in food intake is commonly seen in the mammalian response to stress. Corticotropin-releasing factor (CRF) is a factor that modulates feeding behavior under these stressful conditions. We previously reported that neuropeptide B (NPB) exerts strong synergistic anorectic effects in mice when co-administered with CRF. Tissue distributions of neuropeptide B/W receptor-1 (NPBWR1) and NPB suggest that this neuropeptide system also plays a role in regulating emotion and stress responses. In this study, we examined whether the effects of NPB are altered by endogenous CRFergic tone. During the light period, NPB alone did not show any effect on feeding behavior when administered intracerebroventricularly in mice; however, NPB increased food intake when co-administered with astressin, a potent non-selective CRF receptor antagonist. On the other hand, during the dark period, administration of NPB resulted in a strong inhibition of feeding behavior. The effects were almost completely abolished when astressin was co-administered. Finally, after surgical stress, a semi-chronic stressor, NPB exerted potent anorectic effects on mice. In striking contrast, co-administration of astressin and NPB increased food intake under surgical stress conditions. These data suggest that the effects of NPB are greatly influenced by the endogenous CRFergic tone.