Seasonality of psychopathology and circannual melatonin rhythm

The association of seasonal changes in health and disease has been known for centuries. The prevalence of psychopathological symptoms with seasonal fluctuations and the use of melatonin as a biological marker of circadian and circannual rhythms is well documented. The aim of this work was to study the variability of melatonin secretion between summer and winter in our geographical area (28 degrees N, 16 degrees W) and relate the changes to the level of psychopathology. Ten drug-free, nonsmoker, healthy subjects were studied in summer (August) and winter (December). Blood samples for melatonin assays were collected every hour at night for 5 hr, from 22:00 to 02:00 hr, and next day at noon. Melatonin was assayed by an ELISA technique. Psychopathology was evaluated by means of the 28-item version of the General Health Questionnaire (GHQ-28). All subjects had a circadian rhythm of melatonin secretion in summer and winter. There was a seasonal rhythm with melatonin levels being significantly higher at night in winter than in summer. Melatonin levels at 22:00, 23:00, 24:00 and 01:00 hr and mean melatonin area under the curve (AUC) were significantly higher in winter than in summer. Melatonin AUC increased 80% in winter compared with summer. The GHQ-28 somatic and anxiety subscales and the total GHQ-28 score were significantly higher in winter than summer. Psychopathology scores were significantly and negatively correlated with melatonin production in summer and winter. Our data strongly suggest that melatonin production and psychopathology levels present seasonal fluctuations and these variations should be taken into account when conducting research in this field.     

Evaluation in humans of the effects of radiocellular telephones on the circadian patterns of melatonin secretion, a chronobiological rhythm marker

A decrease in melatonin secretion has been observed in small mammals under exposure to extremely low frequency electromagnetic fields. As there is some concern about possible health effects of the increasing use of radiocellular telephones emitting radiofrequency electromagnetic fields, we examined whether such fields would alter melatonin levels in the human. Volunteers were two groups totalling 38 men, 20-32 yr old. Exposures were to commercially available cellular telephones of the GSM 900 type (Global System for Mobile communication at 900 MHz) or DCS 1800 type (Digital Communication System at 1800 MHz), for 2 hr/day, 5 days/wk, for 4 wk, at their maximum power. Attention of the volunteers was sustained by TV projection of movies. Blood samples were collected hourly during the night and every 3 hr in the daytime. Four sampling sessions were performed at 15-day intervals: before the beginning of the exposure period, at the middle and the end of the exposure period, and 15 days later to evaluate the persistence or late appearance of potential effects. Evaluated parameters were the maximum serum concentration, the time of this maximum, and the area under the curve of the hormone profile. Melatonin circadian profile was not disrupted in 37 young male volunteers submitted to a typical pattern of exposure to the electromagnetic fields generated by two common types of cell phones.     

Exercise and melatonin in humans: reciprocal benefits

The aim of this review is to update the reader as to the association between physical exercise and melatonin, and to clarify how the melatonin rhythm may be affected by different types of exercise. Exercise may act as a zeitgeber, although the effects of exercise on the human circadian system are only now being explored. Depending on the time of the day, on the intensity of light, and on the proximity of the exercise to the onset or decline of the circadian production of melatonin, the consequence of exercise on the melatonin rhythm varies. Moreover, especially strenuous exercise per se induces an increased oxidative stress that in turn may affect melatonin levels in the peripheral circulation because indole is rapidly used to combat free radical damage. On the other hand, melatonin also may influence physical performance, and thus, there are mutually interactions between exercise and melatonin production which may be beneficial.     

Parallel nocturnal secretion of melatonin and testosterone in the plasma of normal men

Abstract: Differences and similarities in the temporal organization of hormone secretion in plasma reflect the activity of CNS pacemakers. One aspect of this activity, the temporal synchronization of the secretion of different hormones is still poorly understood. We report the analysis of melatonin and testosterone plasma concentrations during two nights in 6 normal healthy young men. Blood was collected every 20 min between 2040 and 0640. Plasma testosterone concentrations increased by 1.5- to 2-fold during the second part of the night, and melatonin by 2.5- to 4-fold. In each subject, the individual temporal pattern of melatonin was quite stable over the two nights of sampling, while testosterone profiles showed fluctuations. There was a high degree of parallelism in these two hormones nocturnal secretion. These results, together with previous studies, suggest that melatonin might entrain the nocturnal secretion of testosterone.     

Urinary melatonin: a noninvasive method to follow human pineal function as studied in three experimental conditions

Abstract:  The aim of this study was to examine whether urinary melatonin, rather than urinary 6-sulfatoxymelatonin (aMT6s), can be used as an indicator of diurnally and seasonally changing melatonin secretion. The subjects (n = 15) spent three separate 24-hr periods in a climatic chamber during winter (n = 7) and summer (n = 8). Blood and urine samples were obtained during each period at 2- to 5-hr intervals. Serum melatonin and urinary melatonin and aMT6s were assayed by radioimmunoassay. The serum melatonin levels increased nearly 10-fold from low daytime to high nocturnal values. The mean nocturnal increase of urinary melatonin was 1.7-fold and that of urinary aMT6s was 4.6-fold. Both urinary melatonin and aMT6s correlated significantly with area under the curve melatonin in serum during the night, during the day and throughout the entire 24-hr observation period in all cases. The ratio between urinary melatonin and aMT6s excretion showed significant diurnal variation, being ninefold higher at 16:00 hr than at 07:00 or at 09:00 hr. The ninefold decrease in the urinary melatonin/aMT6s excretion ratio between the evening and the morning may reflect increased liver metabolism of melatonin during the night. Both urinary melatonin and aMT6s are good indicators of melatonin secretion, but the variation is significantly smaller for the former molecule.     

The circadian nature of melatonin secretion in Japanese quail (Coturnix coturnix japonica)

Abstract: Plasma melatonin concentrations were measured in Japanese quail held under different photoperiods and constant darkness (<1 lux). When subjected to LD6:18 (6 hr light: 18 hr darkness), levels rose ～2 hr after lights-off, attained a peak level 8 hr after lights off, and subsequently declined to low daytime levels before the next lights-on signal. This generated a distinct daily rhythm in melatonin secretion with a duration of ～13 h. On exposing quail to a range of photoperiods, containing 6, 9, 11, 12, 13, 15, 18, or 20 hr of light per day, the onset of melatonin secretion remained essentially similar with the rise occurring soon after lights-off. However, the offset of melatonin secretion was suppressed by the light of the next day and thus a much truncated rhythm was produced under long (> 12 hr) photoperiods. Importantly, between night lengths of 4 to 18 hr (i.e., LD 20:4 to LD 6:18) a linear relationship existed between the duration of night-length and secretion of melatonin with the duration increasing by about 0.8 hr for each additional hour of darkness. If quail were released into darkness following a short (LD 6:18) or long (LD 20:4) day schedule, the rhythm persisted for at least two cycles with peaks occurring at about 24 hr intervals. In those quail coming into darkness from long days (LD 20:4), the rhythm of melatonin secretion decompressed rapidly on both sides of the peak, indicating that both the onset and offset of melatonin secretion were suppressed under long days. The endogenous nature of melatonin secretion was tested further by exposing birds to LD 6:30 for 4 cycles and then releasing into darkness. The rhythm in melatonin secretion persisted for at least three cycles before beginning to damp-out. The circadian nature of the rhythm in melatonin secretion was also examined by subjecting quail to T-cycles and then releasing into darkness. Both under the T-cycles and darkness following T-cycle treatments, the phase of the melatonin rhythm was advanced by > 3 hr under T = 27 hr cycles (LD 3:24) compared with T = 24 hr cycles (LD 3:21). This property is consistent with the melatonin oscillator being a circadian rhythm.     

Sirtuins, melatonin and circadian rhythms: building a bridge between aging and cancer

Abstract:  Histone deacetylases (HDAC) have been under intense scientific investigation for a number of years. However, only recently the unique class III HDAC, sirtuins, have gained increasing investigational momentum. Originally linked to longevity in yeast, sirtuins and more specifically, SIRT1 have been implicated in numerous biological processes having both protective and/or detrimental effects. SIRT1 appears to play a critical role in the process of carcinogenesis, especially in age-related neoplasms. Similarly, alterations in circadian rhythms as well as production of the pineal hormone melatonin have been linked to aging and cancer risk. Melatonin has been found act as a differentiating agent in some cancer cells and to lower their invasive and metastatic status. In addition, melatonin synthesis and release occurs in a circadian rhythm fashion and it has been linked to the core circadian machinery genes ( Clock , Bmal1 , Periods , and Cryptochromes ). Melatonin has also been associated with chronotherapy, the timely administration of chemotherapy agents to optimize trends in biological cycles. Interestingly, a recent set of studies have linked SIRT1 to the circadian rhythm machinery through direct deacetylation activity as well as through the nicotinamide adenine dinucleotide (NAD⁺) salvage pathway. In this review, we provide evidence for a possible connection between sirtuins, melatonin, and the circadian rhythm circuitry and their implications in aging, chronomodulation, and cancer.     

Effect of melatonin on motility pattern of small intestine in rats and its inhibition by melatonin receptor antagonist S 22153

Melatonin is synthesized during the night by the pineal gland. Recently, melatonin binding sites have been identified in the gut. Despite few studies, the physiological role of melatonin in gut function remains unclear. The objective of the present study was to investigate the effects of melatonin in the regulation of intestinal motility by using the melatonin receptor antagonist S 22153 in rats. Twenty-four male Wistar rats (400 +/- 25 g) were equipped with intraparietal electrodes along the small intestine. Rats were subjected to a 12:12 hr light:dark schedule. During the dark phase, intestinal migrating motor complexes (MMCs) frequency increased (P < 0.05) by 20% in the duodenum and in the jejunum compared with daylight. This effect is due to a significant reduction in the irregular spiking activity (ISA) of MMCs. Concurrently, at night, the duration of the postprandial motor response is reduced by 30% in the duodenum and 50% in the jejunum and ileum. The administration of S 22153 (2 mg/kg sc) at night suppressed these nocturnal variations and restored the daylight values. In contrast, S 22153 was ineffective during daylight whatever the digestive state. Administration of melatonin (1 mg/kg iv) during the preprandial state, 3 hr after light onset, decreased (-80%) the duration of the ISA of MMCs at the three intestinal levels. During the satiety phase, melatonin administered 10 min before or 15 min after food onset induced the appearance of a transitory preprandial-like motor profile in the entire small intestine. In contrast, when administered at the end of the meal it was ineffective. Preprandial and postprandial melatonin effects were prevented by S 22153 pretreatment. In conclusion, these findings reveal, first, that endogenous melatonin is physiologically involved in the pre- and postprandial changes of intestinal motility at night. Second, exogenous melatonin produces pharmacological effects on pre- and postprandial intestinal motility. In both cases, the action of melatonin corresponds to an inhibition of ISA and a reinforcement of the cyclic MMC pattern.     

The human pineal gland and melatonin in aging and Alzheimer's disease

The pineal gland is a central structure in the circadian system which produces melatonin under the control of the central clock, the suprachiasmatic nucleus (SCN). The SCN and the output of the pineal gland, i.e. melatonin, are synchronized to the 24-hr day by environmental light, received by the retina and transmitted to the SCN via the retinohypothalamic tract. Melatonin not only plays an important role in the regulation of circadian rhythms, but also acts as antioxidant and neuroprotector that may be of importance in aging and Alzheimer's disease (AD). Circadian disorders, such as sleep-wake cycle disturbances, are associated with aging, and even more pronounced in AD. Many studies have reported disrupted melatonin production and rhythms in aging and in AD that, as we showed, are taking place as early as in the very first preclinical AD stages (neuropathological Braak stage I-II). Degeneration of the retina-SCN-pineal axis may underlie these changes. Our recent studies indicate that a dysfunction of the sympathetic regulation of pineal melatonin synthesis by the SCN is responsible for melatonin changes during the early AD stages. Reactivation of the circadian system (retina-SCN-pineal pathway) by means of light therapy and melatonin supplementation, to restore the circadian rhythm and to relieve the clinical circadian disturbances, has shown promising positive results.     

Attenuated nocturnal rise in pineal and serum melatonin in a genetically cardiomyopathic Syrian hamster with a deficient calcium pump

Day and nighttime melatonin production in the pineal gland was compared in normal and cardiomyopathic (polydystrophic) adult male Syrian hamsters. These strains of hamsters were selected for comparison because the cardiomyopathetic hamster displays a deficient transmembrane Ca(2+)-pump in a number of tissues, and intracellular CA2+ concentrations ([Ca2+]i) play a central role in the nocturnal increase in pineal melatonin synthesis. Daytime levels of all constituents measured, i.e., pineal N-acetyltransferase (NAT) activity, pineal and serum melatonin levels, and pineal 5-hydroxytryptophan (5-HTP), serotonin, and 5-hydroxyindole acetic acid (5-HIAA) contents, were comparable in control and dystrophic hamsters. In contrast, the nighttime rises in pineal NAT activity and pineal and serum melatonin levels were significantly attenuated in the dystrophic hamsters. By comparison, the pineal contents of 5-HTP, serotonin, and 5-HIAA were essentially the same in both groups of hamsters with both pineal serotonin and 5-HIAA values exhibiting the usual nighttime drop. It is presumed that the alterations in nocturnal melatonin production in the pineal gland of the cardiomyopathic hamster may relate to a generalized deficiency in the Ca(2+)-pump in pinealocyte plasma membranes, which leads to unusually high [Ca2+]i, causing a depression of NAT activity; this leads to the commensurate decline in pineal and serum melatonin levels. Harderian gland NAT activity and melatonin levels were essentially similar in the two groups of animals, although NAT activity was slightly depressed in the dystrophic hamsters killed during the day. The reduced amounts of intrascapular brown fat in the cardiomyopathic hamster is speculated to be a result of the diminished amount of melatonin produced in these animals.     

Seasonal variations of in vivo and in vitro melatonin production in a passeriform bird, the house sparrow (Passer domesticus)

Melatonin, released from the pineal gland, is an important signal within the circadian pacemaking system of passeriform birds. Until now, seasonal variations in melatonin production have only been examined in a few avian species and the role of melatonin in the regulation of annual rhythms in birds is unclear. We investigated plasma melatonin in a group of house sparrows kept in an outside aviary in spring (March/April), summer (May/June), autumn (September/October), and winter (December/January). The durations of elevated melatonin values mirrored the seasonal changes in night length to a certain degree, the melatonin signal being longest in winter and shortest in summer. Additionally, plasma melatonin peak amplitudes differed significantly among seasons, with highest values in spring and summer and lowest values in winter. Cultured explanted pineal glands obtained from animals in winter and summer showed patterns of in vitro melatonin release comparable to in vivo circulating melatonin with different durations of elevated melatonin and peak amplitude values. These data indicate that the circadian pacemaking system of the house sparrow changes properties seasonally, either as a result of endogenous mechanisms or in response to environmental conditions. These properties are maintained in the pineal gland even after isolation from the animal.     

Perinatal development of circadian melatonin production in domestic chicks

In contrast to the situation in mammals, in which circadian melatonin production by the pineal gland does not begin until some time after birth, the development of pineal gland rhythmicity is an embryonic event in the precocial domestic fowl. A distinct melatonin rhythm was found in 19-d-old chick embryos maintained under light:dark (LD) 16:8. No significant variation in melatonin levels was detected in embryos exposed to LD 8:16. The melatonin rhythm in the pineal gland and plasma of chick embryos incubated for 18 d in LD 12:12 persisted for 2 d in constant darkness indicating that melatonin production is under circadian control at least from the end of embryonic life. A 1-d exposure to a LD cycle during the first postembryonic day was sufficient to entrain the melatonin rhythm, and previous embryonic exposure to either LD or constant darkness (DD) neither modified this rapid synchronization nor did it affect the melatonin pattern during the two subsequent days in DD. It is suggested that, in contrast to the situation in mammals, the avian embryo has evolved its own early circadian melatonin-producing system because, as a consequence of its extrauterine development, it cannot use the system of its mother.     

The association between melatonin and episodic migraine: A pilot network meta-analysis of randomized controlled trials to compare the prophylactic effects with exogenous melatonin supplementation and pharmacotherapy

Although exogenous melatonin supplementation has been suggested to be effective for episodic migraine prophylaxis, there is no conclusive evidence comparing the efficacy of exogenous melatonin supplementation to the other FDA-approved pharmacotherapy for episodic migraine prophylaxis. The aim of the current network meta-analysis (NMA) was to compare the efficacy of exogenous melatonin supplementation in patients with episodic migraine. The randomized placebo-controlled trials or randomized controlled trials (RCTs) incorporating a placebo in the study designs were included in our analyses. All of the NMA procedures were conducted under the frequentist model. The primary outcome was changes in frequency of migraine days and response rate after migraine prophylaxis with melatonin supplementation or pharmacological interventions. We included 25 RCTs in total with 4499 patients (mean age = 36.0 years, mean female proportion = 78.9%). The NMA demonstrated that migraine prophylaxis with oral melatonin 3 mg/d (immediate-release) at bedtime was associated with the greatest improvement in migraine frequency [mean difference = −1.71 days, 95% confidence interval (CI): −3.27 to −0.14 days compared to placebo] and the second highest response rate (odds ratio = 4.19, 95% CI = 1.46 to 12.00 compared to placebo). Furthermore, oral melatonin 3 mg (immediate-release) at bedtime was the most preferred pharmacological intervention among all of the investigated interventions when improvements in migraine frequency, response rate, dropout rate, and rates of any adverse events were taken into account. This pilot NMA suggests the potential prophylactic role of exogenous melatonin supplementation in patients with episodic migraine.     

Differential effects of light wavelength in phase advancing the melatonin rhythm

Shorter wavelength light has been shown to be more effective than longer wavelengths in suppressing nocturnal melatonin and phase delaying the melatonin rhythm. In the present study, different wavelengths of light were evaluated for their capacity to phase advance the saliva melatonin rhythm. Two long wavelengths, 595 nm (amber) and 660 nm (red) and three shorter wavelengths, 470 nm (blue), 497 nm (blue/green), and 525 nm (green) were compared with a no-light control condition. Light was administered via a portable light source comprising two light-emitting diodes per eye, with the irradiance of each diode set at 65 microW/cm(2). Forty-two volunteers participated in up to six conditions resulting in 15 per condition. For the active light conditions, a 2-hr light pulse was administered from 06:00 hr on two consecutive mornings. Half-hourly saliva samples were collected on the evening prior to the first light pulse and the evening following the second light pulse. The time of melatonin onset was calculated for each night and the difference was calculated as a measure of phase advance. The shorter wavelengths of 470, 495 and 525 nm showed the greatest melatonin onset advances ranging from approximately 40-65 min while the longer wavelengths produced no significant phase advance. These results strengthen earlier findings that the human circadian system is more sensitive to the short wavelengths of light than the longer wavelengths.     

Persistence of a plasma melatonin rhythm in constant darkness and its inhibition by constant light in the sleepy lizard, Tiliqua rugosa

This study determined whether a blood plasma melatonin rhythm persists in constant photothermal environments in the sleepy lizard, Tiliqua rugosa. It builds upon an earlier investigation which provided equivocal results as to whether an in vivo melatonin rhythm persists in constant dark (DD) and light (LL) and temperature in this species. Using more frequent sampling points and new assay techniques, the present study showed that the melatonin rhythm persisted for at least 6 days at temperatures of 25 and 33 degrees C in constant dark (DD). The melatonin rhythm, however, was largely eliminated in constant light (LL) at 33 degrees C, thereby contradicting some previous findings in other species of reptiles where melatonin levels were apparently insensitive to an unexpected pulse of light at night. These results demonstrate that the sleepy lizard has a persistent, possibly circadian rhythm of melatonin in DD and constant temperature, and that the rhythm is inhibited by LL and constant temperature. Therefore, the sleepy lizard pineal gland may be an independent oscillator capable of driving the melatonin rhythm and be a transducer of the seasonally changing external photothermal environment.     

Anatomical and cellular localization of melatonin MT1 and MT2 receptors in the adult rat brain

The involvement of melatonin in mammalian brain pathophysiology has received growing interest, but information about the anatomical distribution of its two G‐protein‐coupled receptors, MT₁ and MT₂, remains elusive. In this study, using specific antibodies, we examined the precise distribution of both melatonin receptors immunoreactivity across the adult rat brain using light, confocal, and electron microscopy. Our results demonstrate a selective MT₁ and MT₂ localization on neuronal cell bodies and dendrites in numerous regions of the rat telencephalon, diencephalon, and mesencephalon. Confocal and ultrastructural examination confirmed the somatodendritic nature of MT₁ and MT₂ receptors, both being localized on neuronal membranes. Overall, striking differences were observed in the anatomical distribution pattern of MT₁ and MT₂ proteins, and the labeling often appeared complementary in regions displaying both receptors. Somadendrites labeled for MT₁ were observed for instance in the retrosplenial cortex, the dentate gyrus of the hippocampus, the islands of Calleja, the medial habenula, the suprachiasmatic nucleus, the superior colliculus, the substantia nigra pars compacta, the dorsal raphe nucleus, and the pars tuberalis of the pituitary gland. Somadendrites endowed with MT₂ receptors were mostly observed in the CA3 field of the hippocampus, the reticular thalamic nucleus, the supraoptic nucleus, the inferior colliculus, the substantia nigra pars reticulata, and the ventrolateral periaqueductal gray. Together, these data provide the first detailed neurocytological mapping of melatonin receptors in the adult rat brain, an essential prerequisite for a better understanding of melatonin distinct receptor function and neurophysiology.     

MT1 and MT2 melatonin receptors are expressed in nonoverlapping neuronal populations

Melatonin (MLT) exerts its physiological effects principally through two high‐affinity membrane receptors MT1 and MT2. Understanding the exact mechanism of MLT action necessitates the use of highly selective agonists/antagonists to stimulate/inhibit a given MLT receptor. The respective distribution of MT1 and MT2 within the CNS and elsewhere is controversial, and here we used a “knock‐in” strategy replacing MT1 or MT2 coding sequences with a LacZ reporter. The data show striking differences in the distribution of MT1 and MT2 receptors in the mouse brain: whereas the MT1 subtype was expressed in very few structures (notably including the suprachiasmatic nucleus and pars tuberalis), MT2 subtype receptors were identified within numerous brain regions including the olfactory bulb, forebrain, hippocampus, amygdala and superior colliculus. Co‐expression of the two subtypes was observed in very few structures, and even within these areas they were rarely present in the same individual cell. In conclusion, the expression and distribution of MT2 receptors are much more widespread than previously thought, and there is virtually no correspondence between MT1 and MT2 cellular expression. The precise phenotyping of cells/neurons containing MT1 or MT2 receptor subtypes opens new perspectives for the characterization of links between MLT brain targets, MLT actions and specific MLT receptor subtypes.     

Effect of dose and time of melatonin injections on the diurnal rhythm of immunity in chicken

The effect of daily melatonin injections on the diurnal rhythm of immune parameters was examined in White Leghorn cockerels, kept from hatching in L:D 12:12 conditions. Subcutaneous injections of melatonin were made at the beginning of darkness or 4 h earlier for four weeks starting from one week of life. The melatonin dosage in one group was raised (10, 13, 16, and 20 ng per bird daily, respectively) during four consecutive weeks. The two other doses were 10 and 500 times higher and were increased every week as well. Control birds received equivalent injections of vehicle. Three-week-old chickens were immunized ip with sheep red blood cells and reimmunized one week later. Five-week-old birds were sacrificed during a 24 h period every 4 h. The existence of the diurnal rhythm was evaluated by cosinor analysis. The diurnal rhythm of total white blood cells and serum agglutinins was more dependent on the time of melatonin injections than on the hormone used. The effect of melatonin injections on the level of immune parameters examined was also dependent on the time of sample collection. Results obtained indicate the participation of pineal gland in the regulation of the diurnal rhythm of the examined indices of avian immune system function that exhibit diurnal changes in sensitivity to exogenous melatonin.