Angelman syndrome and melatonin: What can they teach us about sleep regulation.

In 1965, Dr Harry Angelman reported a neurodevelopmental disorder affecting three unrelated children who had similar symptoms: brachycephaly, mental retardation, ataxia, seizures, protruding tongues, and remarkable paroxysms of laughter. Over the past 50 years, the disorder became Angelman's namesake and symptomology was expanded to include hyper-activity, stereotypies, and severe sleep disturbances. The sleep disorders in many Angelman syndrome (AS) patients are broadly characterized by difficulty falling and staying asleep at night. Some of these patients sleep less than 4 hours a night and, in most cases, do not make up this lost sleep during the day-leading to the speculation that AS patients may "need" less sleep. Most AS patients also have severely reduced levels of melatonin, a hormone produced by the pineal gland exclusively at night. This nightly pattern of melatonin production is thought to help synchronize internal circadian rhythms and promote nighttime sleep in humans and other diurnal species. It has been proposed that reduced melatonin levels contribute to the sleep problems in AS patients. Indeed, emerging evidence suggests melatonin replacement therapy can improve sleep in many AS patients. However, AS mice show sleep problems that are arguably similar to those in humans despite being on genetic backgrounds that do not make melatonin. This suggests the hypothesis that the change in nighttime melatonin may be a secondary factor rather than the root cause of the sleeping disorder. The goals of this review article are to revisit the sleep and melatonin findings in both AS patients and animal models of AS and discuss what AS may tell us about the underlying mechanisms of, and interplay between, melatonin and sleep.

The dynamics of GABA signaling: Revelations from the circadian pacemaker in the suprachiasmatic nucleus.

Virtually every neuron within the suprachiasmatic nucleus (SCN) communicates via GABAergic signaling. The extracellular levels of GABA within the SCN are determined by a complex interaction of synthesis and transport, as well as synaptic and non-synaptic release. The response to GABA is mediated by GABAA receptors that respond to both phasic and tonic GABA release and that can produce excitatory as well as inhibitory cellular responses. GABA also influences circadian control through the exclusively inhibitory effects of GABAB receptors. Both GABA and neuropeptide signaling occur within the SCN, although the functional consequences of the interactions of these signals are not well understood. This review considers the role of GABA in the circadian pacemaker, in the mechanisms responsible for the generation of circadian rhythms, in the ability of non-photic stimuli to reset the phase of the pacemaker, and in the ability of the day-night cycle to entrain the pacemaker.

Daily timing of the adolescent sleep phase: Insights from a cross-species comparison.

Adolescence is a time of tremendous adjustment and includes changes in cognition, emotion, independence, social environment, and physiology. One of the most consistent changes exhibited by human adolescents is a dramatic delay in the daily timing of the sleep-wake cycle. This delay is strongly correlated with pubertal maturation and is believed to be influenced by gonadal hormone-induced changes in the neural mechanisms regulating sleep and/or circadian timing. Data from both human and non-human animals indicate that developmental changes in the intrinsic period of the circadian mechanism or its sensitivity to light are not adequate to explain adolescent changes in the daily timing of sleep and wakefulness. Rather, current evidence suggests that pubertal changes in the homeostatic drive to sleep and/or behaviorally induced changes in the amount and/or timing of light exposure permit adolescents to stay up later in the evening and cause them to wake up later in the morning.

Sustained activation of GABAA receptors in the suprachiasmatic nucleus mediates light-induced phase delays of the circadian clock: a novel function of ionotropic receptors.

The suprachiasmatic nucleus (SCN) contains a circadian clock that generates endogenous rhythmicity and entrains that rhythmicity with the day-night cycle. The neurochemical events that transduce photic input within the SCN and mediate entrainment by resetting the molecular clock have yet to be defined. Because GABA is contained in nearly all SCN neurons we tested the hypothesis that GABA serves as this signal in studies employing Syrian hamsters (Mesocricetus auratus). Activation of GABAA receptors was found to be necessary and sufficient for light to induce phase delays of the clock. Remarkably, the sustained activation of GABAA receptors for more than three consecutive hours was necessary to phase-delay the clock. The duration of GABAA receptor activation required to induce phase delays would not have been predicted by either the prevalent theory of circadian entrainment or by expectations regarding the duration of ionotropic receptor activation necessary to produce functional responses. Taken together, these data identify a novel neurochemical mechanism essential for phase-delaying the 'master' circadian clock within the SCN as well as identifying an unprecedented action of an amino acid neurotransmitter involving the sustained activation of ionotropic receptors.

Estradiol acts during a post-pubertal sensitive period to shorten free-running circadian period in male Octodon degus.

The free-running circadian period is approximately 30 min shorter in adult male than in adult female Octodon degus. The sex difference emerges after puberty, resulting from a shortened free-running circadian period in males. Castration before puberty prevents the emergence of the sex difference, but it is not a function of circulating gonadal hormones as such, because castration later in life does not affect free-running circadian period. The aim of this study was to determine whether or not the shortening of the free-running circadian period in male degus results from exposure to gonadal hormones after puberty. We hypothesized that masculinization of the circadian period results from an organizational effect of androgen exposure during a post-pubertal sensitive period. Male degus were castrated before puberty and implanted with capsules filled with dihydrotestosterone (DHT), 17ß-estradiol (E2) or empty capsules at one of three ages: peri-puberty (2-7 months), post-puberty (7-12 months), or adulthood (14-19 months). Long-term exposure to DHT or E2 did not result in a shortened free-running circadian period when administered at 2-7 or 14-19 months of age. However, E2 treatment from 7 to 12 months of age decreased the free-running circadian period in castrated males. This result was replicated in a subsequent experiment in which E2 treatment was limited to 8-12 months of age. E2 treatment at 7-12 months of age had no effect on the free-running circadian period in ovariectomized females. Thus, there appears to be a post-pubertal sensitive period for sexual differentiation of the circadian system of degus, during which E2 exposure decreases the free-running circadian period in males. These data demonstrate that gonadal hormones can act during adolescent development to permanently alter the circadian system.

Period gene expression in the diurnal degu (Octodon degus) differs from the nocturnal laboratory rat (Rattus norvegicus).

Recent data suggest that both nocturnal and diurnal mammals generate circadian rhythms using similarly phased feedback loops involving Period genes in the suprachiasmatic nuclei (SCN) of the hypothalamus. These molecular oscillations also exist in the brain outside of the SCN, but the relationship between SCN and extra-SCN oscillations is unclear. We hypothesized that a comparison of "diurnal" and "nocturnal" central nervous system Per rhythms would uncover differences in the underlying circadian mechanisms between these two chronotypes. Therefore, this study compared the 24-h oscillatory patterns of Per1 and Per2 mRNA in the SCN and putative striatum and cortex of Octodon degus (degu), a diurnal hystricognath rodent, with those of the nocturnal laboratory rat, Rattus norvegicus. The brains of adult male degus and rats were collected at 2-h intervals across 24 h in entrained light-dark and constant darkness conditions, and sections were analyzed via in situ hybridization. In the SCN, degu Per1 and Per2 hybridization signal exhibited 24-h oscillatory patterns similar in phasing to those seen in other rodents, with peaks occurring during the light period and troughs during the dark period. However, Per1 remained elevated for five fewer hours in the degu than in the rat, and Per2 remained elevated for two fewer hours in the degu. In brain areas outside of the SCN, the phase of Per2 hybridization signal rhythms in the degu were 180 degrees out of phase with those found in the rat, and Per1 hybridization signal lacked significant rhythmicity. These results suggest that, while certain basic components of the transcriptional-translational feedback loop generating circadian rhythms are similar in diurnal and nocturnal mammals, there are variations that may reflect adaptations to circadian niche.

Gonadal hormone effects on entrained and free-running circadian activity rhythms in the developing diurnal rodent Octodon degus.

The slowly maturing, long-lived rodent Octodon degus (degu) provides a unique opportunity to examine the development of the circadian system during adolescence. These studies characterize entrained and free-running activity rhythms in gonadally intact and prepubertally gonadectomized male and female degus across the first year of life to clarify the impact of sex and gonadal hormones on the circadian system during adolescence. Gonadally intact degus exhibited a delay in the phase angle of activity onset (Psi(on)) during puberty, which reversed as animals became reproductively competent. Gonadectomy before puberty prevented this phase delay. However, the effect of gonadal hormones during puberty on Psi(on) does not result from changes in the period of the underlying circadian pacemaker. A sex difference in Psi(on) and free-running period (tau) emerged several months after puberty; these developmental changes are not likely to be related, since the sex difference in Psi(on) emerged before the sex difference in tau. Changes in the levels of circulating hormones cannot explain the emergence of these sex differences, since there is a rather lengthy delay between the age at which degus reach sexual maturity and the age at which Psi(on) and tau become sexually dimorphic. However, postnatal exposure to gonadal hormones is required for sexual differentiation of Psi(on) and tau, since these sex differences were absent in prepubertally gonadectomized degus. These data suggest that gonadal hormones modulate the circadian system during adolescent development and provide a new model for postpubertal sexual differentiation of a central nervous system structure.

Pubertal development of sex differences in circadian function: an animal model.

UNLABELLED: The development of adult circadian function, particularly sexual dimorphism of function, has been well studied only in rapidly developed rodents. In such species development is complete by weaning. Data from adolescent humans suggest that significant development occurs during the pubertal period. We hypothesized that a more slowly developing rodent might better mimic the changes in circadian function around puberty in humans and allow us to determine the underlying neural changes. Entrained and free-running circadian rhythms were analyzed and correlated with pubertal development in male and female Octodon degus (degu) that remained gonadally intact or were gonadectomized at weaning. Brains were collected during development to measure androgen and estrogen receptors in the suprachiasmatic nuclei (SCN) Adult circadian period does not develop until 10-12 months of age in degus, long after the onset of gonadal maturation (3-5 months). The timing of circadian period maturation correlates with the appearance of steroid receptors in the SCN. Changes in free-running rhythms only occurred in gonadally intact degus. Adult phase angles of activity onset develop between 2 and 3 months of age (comparing results of two experiments), soon after the onset of pubertal changes. CONCLUSION: The development of sexually dimorphic adult circadian period occurs after gonadal puberty is complete and requires the presence of gonadal steroids. The delay in development until after gonadal puberty is likely due to the delayed appearance of steroid receptors in the SCN. Phase is not sexually dimorphic and changes in the absence of steroid hormones.