Orexin-A is composed of a highly conserved C-terminal and a specific, hydrophilic N-terminal region, revealing the structural basis of specific recognition by the orexin-1 receptor.

Orexins-A and B, also called hypocretins-1 and 2, respectively, are neuropeptides that regulate feeding and sleep-wakefulness by binding to two orphan G protein-coupled receptors named orexin-1 (OX(1)R) and orexin-2 (OX(2)R). The sequences and functions of orexins-A and B are similar to each other, but the high sequence homology (68%) is limited in their C-terminal half regions (residues 15-33). The sequence of the N-terminal half region of orexin-A (residues 1-14), containing two disulfide bonds, is very different from that of orexin-B. The structure of orexin-A was determined using two-dimensional homonuclear and (15)N and (13)C natural abundance heteronuclear NMR experiments. Orexin-A had a compact conformation in the N-terminal half region, which contained a short helix (III:Cys6-Gln9) and was fixed by the two disulfide bonds, and a helix-turn-helix conformation (I:Leu16-Ala23 and II:Asn25-Thr32) in the remaining C-terminal half region. The C-terminal half region had both hydrophobic and hydrophilic residues, which existed on separate surfaces to provide an amphipathic character in helices I and II. The nine residues on the hydrophobic surface are also well conserved in orexin-B, and it was reported that the substitution of each of them with alanine resulted in a significant drop in the functional potency at the receptors. Therefore, we suggest that they form the surface responsible for the main hydrophobic interaction with the receptors. On the other hand, the residues on the hydrophilic surface, together with the hydrophilic residues in the N-terminal half region that form a cluster, are known to make only small contributions to the binding to the receptors through similar alanine-scan experiments. However, since our structure of orexin-A showed that large conformational and electrostatical differences between orexins-A and B were rather concentrated in the N-terminal half regions, we suggest that the region of orexin-A is important for the preference for orexin-A of OX(1)R.     

Dream Content in Patients with Narcolepsy: Preliminary Findings

The present study investigated dream recall and dream content in patients with narcolepsy. Compared to healthy controls, patients with narcolepsy reported higher dream recall frequency and more negatively toned and bizarre dreams, confirming earlier findings.     

Metabolic state signalling through central hypocretin/orexin neurons

Hypothalamic neurons that produce the peptide transmitters hypocretins/orexins have attracted much recent attention. They provide direct and predominantly excitatory inputs to all major brain areas except the cerebellum, with the net effect of stimulating wakefulness and arousal. These inputs are essential for generating sustained wakefulness in mammals, and defects in hypocretin signalling result in narcolepsy. In addition, new roles for hypocretins/orexins are emerging in reward-seeking, learning, and memory. Recent studies also indicate that hypocretin/orexin neurons can alter their intrinsic electrical activity according to ambient fluctuations in the levels of nutrients and appetite-regulating hormones. These intriguing electrical responses are perhaps the strongest candidates to date for the elusive neural correlates of after-meal sleepiness and hunger-induced wakefulness. Hypocretin/orexin neurons may thus directly translate rises and falls in body energy levels into different states of consciousness.     

Clinical significance of social jetlag in patients with excessive daytime sleepiness

ABSTRACT

Social jetlag has recently attracted attention as the circadian misalignment between biological and social clocks. We aimed to examine social jetlag and its effect on daytime sleepiness and daily functions in patients with narcolepsy, behaviorally induced insufficient sleep syndrome (BIISS) and delayed sleep-wake phase disorder (DSPD). The levels of social jetlag (SJL_mid) and sleep-corrected social jetlag (SJL_sc) were calculated for each patient, and the effect of these social jetlag-related parameters on daytime sleepiness and daily functions were examined. Objective sleepiness measured by the mean sleep latency in the multiple sleep latency test, subjective sleepiness assessed by the Epworth sleepiness scale (ESS), health-related quality of life (HRQoL) assessed by the SF-8 health survey, and incidences of mistakes in daily activities, traffic accidents and near-miss events related to daytime sleepiness were compared among the narcolepsy (n = 39), BIISS (n = 87) and DSPD (n = 28) groups. Both SJL_mid and SJL_sc showed a negative correlation with physical HRQoL in patients with narcolepsy and a positive correlation with the ESS score in patients with DSPD. In patients with BIISS, SJL_sc reflected sleep loss rather than circadian misalignment; moreover, SJL_sc was not associated with daytime sleepiness and daily functions. Social jetlag was not associated with incidences of mistakes in daily activities, traffic accidents and near-miss events.

The state of social jetlag and its association with daily functions differed among the narcolepsy, BIISS and DSPD groups. Social jetlag represented sleep debt in BIISS, circadian misalignment in narcolepsy and both in DSPD. Our results thus show that the clinical manifestations and significance of social jetlag differ depending on the underlying sleep disorders.     

Evaluation of (R)-(-)-α-methoxy phenyl acetic acid as a chiral shift reagent for resolution and determination of R and S enantiomers of modafinil in bulk drugs and formulations by 1H NMR spectroscopy

(R)-(-)-α-Methoxy phenyl acetic acid, (S)-(-)-1,1'-(2-naphthol), and (R)-(+)-α-methoxy-α-trifluoromethyl phenyl acetic acid were evaluated as chiral shift reagents (CSRs) for (1)H NMR spectroscopic resolution and determination of R and S enantiomers of modafinil (MDL) in bulk drugs and formulations. Effects of the nature of CSR and the weight ratio of substrate to shift reagent on enantiomeric discrimination were investigated. Intramolecular and intermolecular hydrogen bonding interactions between the drug and the CSR seem to be the driving force for desired resolution. A mechanism was proposed to explain the interactions between (R, S)-enantiomers of MDL and (R)-(-)-α-methoxy phenyl acetic acid. The method was validated and applied successfully to determine the enantiomeric purity of MDL in tablet formulations.     

Hypocretin Deficiency in Narcoleptic Humans Is Associated with Abdominal Obesity

Objective: To determine the prevalence of obesity among patients with narcolepsy, to estimate associated long‐term health risks on the basis of waist circumference, and to distinguish the impact of hypocretin deficiency from that of increased daytime sleepiness (i.e., reduced physical activity) on these anthropometric measures. Research Methods and Procedures: A cross‐sectional, case‐control study was conducted. Patients with narcolepsy (n = 138) or idiopathic hypersomnia (IH) (n = 33) were included. Age‐matched, healthy members of the Dutch population (Monitoring Project on Risk Factors for Chronic Diseases and Doetinchem Project; n = 10, 526) were used as controls. BMI and waist circumference were determined. Results: Obesity (BMI ≥ 30 kg/m²) and overweight (BMI 25 to 30 kg/m²) occurred more often among narcolepsy patients [prevalence: 33% (narcoleptics) vs. 12.5% (controls) and 43% (narcoleptics) vs. 36% (controls), respectively; both p < 0.05]. Narcoleptics had a larger waist circumference (mean difference 5 ± 1.4 cm, p < 0.001). The BMI of patients with IH was significantly lower than that of narcolepsy patients (25.6 ± 3.6 vs. 28.5 ± 5.4 kg/m²; p = 0.004). Discussion: Overweight and obesity occur frequently in patients with narcolepsy. Moreover, these patients have an increased waist circumference, indicating excess fat storage in abdominal depots. The fact that patients with IH had a lower BMI than narcoleptics supports the notion that excessive daytime sleepiness (i.e., inactivity) cannot account for excess body fat in narcoleptic patients.     

Behavioral and sleep/wake characteristics of mice lacking norepinephrine and hypocretin

We investigated the interaction between norepinephrine (NE) and orexin/hypocretin (Hcrt) in the control of sleep behavior and narcoleptic symptoms by creating mice that were deficient in both neurotransmitters. Mice with a targeted disruption of the dopamine beta-hydroxylase (Dbh) gene (deficient in NE and epinephrine) or the Hcrt gene were bred to generate double knockouts (DKOs), each single KO (Dbh-KO and Hcrt-KO), and control mice. The duration of wake, non-rapid eye movement (NREM) and REM sleep were monitored by electroencephalogram (EEG)/electromyogram (EMG) recording over a 24-h period, and the occurrence of behavioral arrests was monitored by video/EEG recording for 4 h. Overall, there was very little interaction between the two genes; for most parameters that were measured, the DKO mice resembled either Dbh-KO or Hcrt-KO mice. REM sleep was increased in both DKO and Hcrt-KO mice at night relative to the other groups, but DKO mice had significantly more REM sleep during the day than the other three groups. Sleep latency in response to saline or amphetamine injections was reduced in Dbh-KO and DKO mice relative to other groups. Behavioral arrests, that are frequent in Hcrt-KO mice, were not exacerbated in DKO mice.     

Immortal orexin cell transplants restore motor-arousal synchrony during cataplexy

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