Nocturnal enhancement of plasma melatonin could be suppressed by benzodiazepines in humans

Plasma melatonin levels were determined every 20 and 30 min for 24 hours on the last day of repeated oral administrations (1 or 2 mg a day for 8 or 9 days) of a benzodiazepine derivative (450191-s), which is known to be metabolized to active benzodiazepines after administration. In one of the two subjects, the nocturnal enhancement of plasma melatonin which was obvious on a control day with placebo was diminished almost completely. In the other subject, observed were not only the diminishment of its nocturnal enhancement but also its increase during the daytime almost to the nocturnal levels on a control day, which may indicate a rebound increase in melatonin synthesis or a shift in its day-night rhythmicity. Such suppressing effects of benzodiazepines on the nocturnal plasma melatonin levels were also examined in the case of a single administration of 2 mg of 450191-s or flunitrazepam in the second series of experiments. Even a single flunitrazepam seemed to have lowered nocturnal plasma melatonin levels, which then recovered to the usual levels following the administration of 5 mg of a benzodiazepine antagonist, Ro 15-1788, given 6 hours after the flunitrazepam. However, single 450191-s did not show any remarkable effects. Thus, it has been suggested that benzodiazepines could suppress the nocturnal levels of plasma melatonin or shift its day-night rhythmicity at least when administered repeatedly. The possible action site of benzodiazepines may be the central nervous system, since melatonin synthesis has been though to be under strongly regulated by the central nervous pathway from the retina to the pineal body. Therefore, these effects of benzodiazepines may provide a method for investigating the physiological role of melatonin and its day-night rhythmicity as well as to further clarify the system regulating melatonin synthesis in humans.     

Sleep enhances nocturnal plasma ghrelin levels in healthy subjects

Ghrelin, an endogenous ligand of the growth hormone secretagogue receptor, has been shown to promote slow-wave sleep (SWS, non-REM sleep stages 3 and 4). Plasma levels of ghrelin are dependent on food intake and increase in sleeping subjects during the early part of the night. It is unknown whether sleep itself affects ghrelin levels or whether circadian networks are involved. Therefore, we studied the effect of sleep deprivation on nocturnal ghrelin secretion. In healthy male volunteers, plasma levels of ghrelin, cortisol, and human growth hormone (hGH) were measured during two experimental sessions of 24 h each: once when the subjects were allowed to sleep between 2300 and 0700 and once when they were kept awake throughout the night. During sleep, ghrelin levels increased during the early part of the night and decreased in the morning. This nocturnal increase was blunted during sleep deprivation, and ghrelin levels increased only slightly until the early morning. Ghrelin secretion during the first hours of sleep correlated positively with peak hGH concentrations. We conclude that the nocturnal increase in ghrelin levels is more likely to be caused by sleep-associated processes than by circadian influences. During the first hours of sleep, ghrelin might promote sleep-associated hGH secretion and contribute to the promotion of SWS.     

Exercise elicits phase shifts and acute alterations of melatonin that vary with circadian phase

To examine the immediate phase-shifting effects of high-intensity exercise of a practical duration (1 h) on human circadian phase, five groups of healthy men 20-30 yr of age participated in studies involving no exercise or exposure to morning, afternoon, evening, or nocturnal exercise. Except during scheduled sleep/dark and exercise periods, subjects remained under modified constant routine conditions allowing a sleep period and including constant posture, knowledge of clock time, and exposure to dim light intensities averaging (+/-SD) 42 +/- 19 lx. The nocturnal onset of plasma melatonin secretion was used as a marker of circadian phase. A phase response curve was used to summarize the phase-shifting effects of exercise as a function of the timing of exercise. A significant effect of time of day on circadian phase shifts was observed (P < 0.004). Over the interval from the melatonin onset before exercise to the first onset after exercise, circadian phase was significantly advanced in the evening exercise group by 30 +/- 15 min (SE) compared with the phase delays observed in the no-exercise group (-25 +/- 14 min, P < 0.05). Phase shifts in response to evening exercise exposure were attenuated on the second day after exercise exposure and no longer significantly different from phase shifts observed in the absence of exercise. Unanticipated transient elevations of melatonin levels were observed in response to nocturnal exercise and in some evening exercise subjects. Taken together with the results from previous studies in humans and diurnal rodents, the current results suggest that 1) a longer duration of exercise exposure and/or repeated daily exposure to exercise may be necessary for reliable phase-shifting of the human circadian system and that 2) early evening exercise of high intensity may induce phase advances relevant for nonphotic entrainment of the human circadian system.     

Plasma testosterone during nocturnal sleep in normal men

Plasma testosterone was measured by a competitive protein binding procedure at 10 to 20 minute intervals in five normal adult men during two nights of sleep. Blood samples were obtained by means of an indwelling venous catheter while sleep was monitored polygraphically. There were 1–4 abrupt elevations of plasma testosterone concentration per night in each of the subjects with an average increase of 244 ng/100 ml ± 45.5 (SE) or 59% above the values present at the onset of the episode. The fluctuations in plasma testosterone were superimposed on a nocturnal rise of the hormone observed in seven of the nights. The average of all samples taken during each hour period through the ten nights revealed a highly significant (P<0.001) nocturnal increase in plasma testosterone. The findings did not support the existence of a relation between REM sleep and an increase in testosterone levels.     

Entrainment of rhythmic melatonin secretion in man to a 12-hour phase shift in the light/dark cycle

Daily rhythms in melatonin secretion were monitored in four healthy adult males by measuring the melatonin contents of sequential 4-hour urine specimens and of plasma samples collected at 12-hour intervals, or, in one subject, continuously for 24 hours. All subjects exhibited similar diurnal rhythms, with peak urinary melatonin excretion rates and blood melatonin levels occurring during the daily period of darkness and sleep. When the daily light/dark regimen was phase-shifted by 180°, the plasma and urinary melatonin rhythms required 5–7 days (depending on the subject) to re-entrain to the new schedule. Simultaneous measurements of plasma melatonin levels and melatonin excretion rates indicate that urinary melatonin reflects, with remarkable fidelity, circulating melatonin levels.     

Repeated exposures to daytime bright light increase nocturnal melatonin rise and maintain circadian phase in young subjects under fixed sleep schedule

Effects of two different light intensities during daytime were examined on human circadian rhythms in plasma melatonin, core body temperature, and wrist activity under a fixed sleep schedule. Sleep qualities as indicated by polysomnography and subjective sleepiness were also measured. In the first week, under dim light conditions ( approximately 10 lx), the onset and peak of nocturnal melatonin rise were significantly delayed, whereas the end of melatonin rise was not changed. The peak level of melatonin rise was not affected. As a result, the width of nocturnal melatonin rise was significantly shortened. In the second week, under bright light conditions ( approximately 5,000 lx), the phases of nocturnal melatonin rise were not changed further, but the peak level was significantly increased. Core body temperature at the initial sleep phase was progressively elevated during the course of dim light exposure and reached the maximum level at the first night of bright light conditions. Subjective sleepiness gradually declined in the course of dim light exposure and reached the minimum level at the first day of bright light. These findings indicate that repeated exposures to daytime bright light are effective in controlling the circadian phase and increasing the peak level of nocturnal melatonin rise in plasma and suggest a close correlation between phase-delay shifts of the onset of nocturnal melatonin rise or body temperature rhythm and daytime sleepiness.     

Nocturnal plasma levels of melatonin in quails (Coturnix japonica) injected with l-5-hydroxy-tryptophan.

This study aimed to demonstrate the influence of the systemic administration of l-5-hydroxy-tryptophan (L-HTP) on the plasma levels of melatonin during the dark period in quails. Throughout daylight, the plasma levels of melatonin did not differ significantly, oscillating between 110.2 +/- 15.8 pg.mL(-1) and 157.4 +/- 34.8 pg.mL(-1), from 8 to 16 hours. L-HTP (25 mg.kg(-1), through the intracelomic route) administered at 18 hours lessened significantly the nocturnal increase of the plasma levels of melatonin (controls, 327.3 +/- 20.1 and 315.8 +/- 20.9 pg.mL(-1) vs. 242.1 +/- 24.8 and 217.5 +/- 21 pg.mL(-1), respectively, at 20 and 24 hours, P < 0.05). The results obtained showed that the administration of LHTP reduced the nocturnal melatonin release, possibly by bringing about an increase in serotonin synthesis and synaptic release in the pineal. Therefore, the serotoninergic transmission from the raphe towards the pineal would constitute a mechanism of modulation of the synthesis and melatonin release in quails.     

Analysis of trough and peak of plasma melatonin circadian rhythm in ewes

This study evaluates the pattern of plasma melatonin during the trough and the peak of its daily rhythm. Blood samples from 8 ewes were collected every 3 h for a 48-h period. On the third day, blood samples were collected from 10:00 to 13:00 (trough) and from 20:00 to 23:00 (peak) every 20 min. Our results showed a robust daily rhythm of melatonin in both days of monitoring, with nocturnal acrophase. During the trough, a significant decrease was observed starting from the 10:40 with a progressive decrease about every 40 min. During the peak of the plasma melatonin daily rhythm, an increase was observed starting from the 20:40 with a progressive increase about every 40 min. These data could be taken in consideration to monitor the plasma melatonin variations during the 24 h, and for the administration of melatonin for breeding in ewes.     

Increased REM sleep associated with melatonin deficiency after pinealectomy: a case study

The objectives of the investigation were to assess hypersomnia, which progressively appeared in a young patient after a pinealectomy, chemotherapy, and radiotherapy for a typical germinoma, as well as the potential benefit of melatonin administration in the absence of its endogenous secretion. 24 h ambulatory polysomnography and the Multiple Sleep Latency Test (MSLT) were performed; in addition, daily plasma melatonin, cortisol, growth hormone, prolactin, and rectal temperature profiles were determined before and during melatonin treatment (one 2 mg capsule given nightly at 21:00 h for 4 weeks). MSLT showed abnormal sleep latency and two REM sleep onsets. Nighttime total sleep duration was lengthened, mainly as a consequence of an increased REM sleep duration. These parameters were slightly modified by melatonin replacement. Plasma melatonin levels, which were constantly nil in the basal condition, were increased to supraphysiological values with melatonin treatment. The plasma cortisol profile showed nycthemeral variation within the normal range, and the growth hormone profile showed supplementary diurnal peaks. Melatonin treatment did not modify the secretion of either hormone. The plasma prolactin profile did not display a physiological nocturnal increase in the basal condition; however, it did during melatonin treatment, with the rise coinciding with the nocturnal peak of melatonin concentration. A 24 h temperature rhythm of normal amplitude was persistent, though the mean level was decreased and the rhythm was dampened during melatonin treatment. The role of radiotherapy on the studied parameters cannot be excluded; the findings of this case study suggest that the observed hypersomnia is not the result of melatonin deficiency alone. Overall, melatonin treatment was well tolerated, but the benefit on the sleep abnormality, especially on daytime REM sleep, was minor, requiring the re-introduction of modafinil treatment.     

Increased REM Sleep Associated with Melatonin Deficiency after Pinealectomy: A Case Study

The objectives of the investigation were to assess hypersomnia, which progressively appeared in a young patient after a pinealectomy, chemotherapy, and radiotherapy for a typical germinoma, as well as the potential benefit of melatonin administration in the absence of its endogenous secretion. 24 h ambulatory polysomnography and the Multiple Sleep Latency Test (MSLT) were performed; in addition, daily plasma melatonin, cortisol, growth hormone, prolactin, and rectal temperature profiles were determined before and during melatonin treatment (one 2 mg capsule given nightly at 21:00 h for 4 weeks). MSLT showed abnormal sleep latency and two REM sleep onsets. Nighttime total sleep duration was lengthened, mainly as a consequence of an increased REM sleep duration. These parameters were slightly modified by melatonin replacement. Plasma melatonin levels, which were constantly nil in the basal condition, were increased to supraphysiological values with melatonin treatment. The plasma cortisol profile showed nycthemeral variation within the normal range, and the growth hormone profile showed supplementary diurnal peaks. Melatonin treatment did not modify the secretion of either hormone. The plasma prolactin profile did not display a physiological nocturnal increase in the basal condition; however, it did during melatonin treatment, with the rise coinciding with the nocturnal peak of melatonin concentration. A 24 h temperature rhythm of normal amplitude was persistent, though the mean level was decreased and the rhythm was dampened during melatonin treatment. The role of radiotherapy on the studied parameters cannot be excluded; the findings of this case study suggest that the observed hypersomnia is not the result of melatonin deficiency alone. Overall, melatonin treatment was well tolerated, but the benefit on the sleep abnormality, especially on daytime REM sleep, was minor, requiring the re‐introduction of modafinil treatment.     

The effects of nocturnal life on endocrine circadian patterns in healthy adults

We observed the 24-hour patterns of endocrine in medical students who lived either a diurnal life or nocturnal life. Nocturnal life was designed by skipping their breakfast but consuming much (>50% of their daily food intake) in the evening and at night with the sleep from 0130 h to 0830 h the next morning. After 3 weeks in the experimental life, the 24-hour plasma concentrations of melatonin, leptin, glucose and insulin were measured every three hours. Both plasma melatonin and leptin showed peaks at 0300 h in the diurnal lifestyle group, and the night peaks decreased in the nocturnal lifestyle group. The changes in the patterns of melatonin and leptin were highly consistent with that of night-eating syndrome (NES). Plasma glucose increased after all meals in both groups. Its concentration maintained a high level in the nocturnal lifestyle group between midnight and early morning while insulin secretion decreased markedly during this period. Furthermore, the strong association between glucose and insulin in the diurnal lifestyle group after meals was damaged in the nocturnal lifestyle group. It was suggested that nocturnal life leads to the impairment of insulin response to glucose. Taking these results together, nocturnal life is likely to be one of the risk factors to health of modern people, including NES, obesity and diabetes.     

The effects of bright light and nighttime melatonin administration on cardiac activity

Although melatonin has an important physiological role in the facilitation of sleep, its precise mechanism of action is not clear. To investigate the potential contribution of melatonin to influence cardiac autonomic activity in the evening, 16 young healthy subjects participated in a repeated measures design where cardiac autonomic activity, heart rate and blood pressure were examined during three experimental conditions. An initial baseline condition involved dim light exposure (< 10 lux), permitting the normal nocturnal rise in endogenous melatonin. In other sessions, subjects were exposed to bright light (> 3000 lux) to suppress melatonin secretion and administered a placebo or melatonin (5 mg) capsule at the estimated time of increase in endogenous melatonin (wake time + 14 hours). Heart rate, pre-ejection period (a measure of cardiac sympathetic activity) and respiratory sinus arrhythmia (a measure of parasympathetic activity) were not significantly altered in response to the three melatonin levels. While melatonin had no effect on diastolic blood pressure, systolic blood pressure was maximally decreased by 6 +/- 1.93 mmHg (mean +/- SEM, p < 0.005) 150 minutes after exogenous melatonin. The results indicate that melatonin does not directly modulate cardiac autonomic activity, but may rather act directly on the cardiovascular system.     

Twenty-Four-Hour Prolactin Profiles in Night Workers

In addition to sleep processes, it has been suggested that an intrinsic circadian rhythmicity is involved in the temporal organization of prolactin (PRL) secretion. Eight night workers were studied to determine whether the PRL rhythm is adapted to their rest-activity schedule and whether this provides evidence in favor of an endogenous clock-driven component. Ten day-active subjects, sleeping once during the night and once after an 8-h delay in their sleep period, were used as a control group. Plasma PRL, body temperature, and plasma melatonin were measured at 10-min intervals. Twenty-four-hour PRL profiles did not differ between night workers sleeping as usual during the daytime and day-active subjects submitted to an abrupt sleep shift to daytime. For the two groups of subjects a transient PRL peak, similar in size and time of occurrence, was observed during the night. Melatonin, a strong marker of the primary circadian oscillator, displayed a phase shift that differed widely among night workers. Body temperature, on the other hand, was found to be more regularly adapted despite the persistence of a small decrease or leveling off during the night. Although no relationship was found between the melatonin increase and the nocturnal PRL peak, a concomitance with this transient temperature decrease could be demonstrated. The persistence of this PRL peak in night workers raises the question of its significance. (Chronobiology International, 13(4), 283-293, 1996)     

Change in sleep state of the elderly before and after cataract surgery

It seems likely that the influences of light upon circadian rhythms will decrease with aging, particularly those rhythms that are more influenced by light with a higher color temperature and richer in short wavelengths. More specifically, cataract patients' optical systems transmit light poorly, especially the shorter wavelengths that affect the circadian system more. The present study investigated melatonin secretion profiles and sleep patterns before and after cataract surgery. Fifteen subjects were studied for 3 consecutive weekdays before, and one month after, their cataract surgery. UV-cutting intra-ocular lenses were used for patients after surgery. No statistically significant differences between before and after surgery were observed in the amount of light received and the amount of activity. This means that there were no significant changes in their lifestyle during the experimental period. Considering the group as a whole, no significant differences were present in melatonin secretion, sleep parameters, or sleepiness before and after the surgery. However, individual subjects responded differently. The subjects showed a negative correlation between the wake-up (p=0.067) or retiring times (p=0.017) and sleep efficiency after surgery. The amount of light received during the nighttime influenced sleep more significantly than during the daytime.     

Melatonin concentrations in the two jugular veins, and relationship with the seasonal reproductive activity in goats

The authors investigated whether melatonin concentrations vary between the two jugular veins and whether absolute (nocturnal) or relative (nocturnal/diurnal ratio) plasma melatonin concentrations are associated with seasonal reproductive activity measured by oestrus or ovulatory activity in Payoya goats. Thirty-two adult Payoya goats were penned under natural photoperiod. Oestrus activity was tested daily using aproned males—twice a week plasma was sampled for progesterone. Melatonin plasma concentrations were studied at each equinox and solstice of the year in jugular samples taken simultaneously by venipuncture. Nocturnal and diurnal plasma melatonin concentrations from each jugular vein were assessed in 3 and 2 plasma samples per goat, respectively, taken at hourly intervals in each period. No differences in melatonin concentrations between the two veins were observed, but there was a significant interaction (P < 0.001) between jugular vein and animal in nocturnal melatonin concentrations. There was no effect of sampling period on melatonin concentrations and the coefficient of correlation between sampling periods was very high. The analyses performed indicated that neither absolute nor relative melatonin concentrations were related with the dates of onset or end of ovulatory/oestrus activity. Therefore, we concluded that in goats (1) melatonin concentrations are highly variable between jugular veins in the same individual but not in the general population, (2) melatonin concentrations are highly repeatable for each individual, and (3) absolute and relative amplitudes of melatonin concentrations are not linked to the seasonal breeding activity in Mediterranean goats.     

Nocturnal variation of prolactin secretion in the Mouflon (Ovis gmelini musimon) and domestic sheep (Ovis aries): seasonal changes

Seasonal changes in nocturnal prolactin secretion and their relationship with melatonin secretion were monitored in wild (Mouflon, Ovis gmelini musimon) and domesticated sheep (breed Manchega, Ovis aries). Two groups of eleven adult females each, were maintained outdoors under natural photoperiod. Plasma concentrations of prolactin and melatonin were determined during the summer and winter solstices and the autumn and spring equinoxes. Blood samples were collected every 3h during the night hours, and 1h before and after the onset of darkness and sunrise. Maximum mean plasma concentrations of prolactin during the dark-phase in Mouflons were observed in the summer solstice, (P<0.001) and in the summer solstice and spring equinox in Manchega ewes (P<0.001). Mean plasma concentrations of prolactin were higher in the wild species (P<0.001) during the summer solstice. In contrast, during the spring equinox, mean levels of prolactin were higher in Manchega ewes than in Mouflons (P<0.05). Plasma prolactin concentrations showed a nocturnal rhythm in both breeds, with seasonal variations (P<0.001). The increase in plasma melatonin levels during the first hour after sunset was accompanied to increasing concentrations of PRL 1h after the onset of darkness, only in the autumn and spring equinox for the Mouflon, and in the summer solstice and spring equinox for the Manchega ewes. In Mouflons, the fall of plasma PRL concentrations about the middle dark-phase in all the periods studied, coincided with high levels of melatonin. A similar relation was observed in Manchega ewes only in the winter solstice and spring equinox. The current study shows that the nocturnal rhythm of prolactin secretion exhibits seasonal variation; differences in the patterns of prolactin secretion between Mouflon and Manchega sheep are taken to represent the effects of genotype.     

Effects of melatonin feeding on smoltification in masu salmon (Oncorhynchus masou)

Influences of photoperiod on plasma melatonin profiles and effects of melatonin administration on long-day-induced smoltification in masu salmon (Oncorhynchus masou) were investigated in order to reveal the roles of melatonin in the regulation of smoltification in salmonids. Under light-dark (LD) cycles, plasma melatonin levels exhibited daily variation, with higher values during the dark phase than during the light phase. The duration of nocturnal elevation under short photoperiod (LD 8:16) was longer than that under long photoperiod (LD 16:8). Melatonin feeding (0.01, 0.1 and 1 mg/kg body weight) elevated plasma levels of melatonin in a dose-dependent manner for at least 7 h but not for 24 h. When masu salmon reared under short photoperiod were exposed to long photoperiod (LD 16:8) and fed melatonin (1 mg/kg body weight) 7 hours before the onset of darkness, a significantly smaller proportion of smolts appeared in the melatonin-fed group after 32 days than in the control group. However, after 59 days of the treatment, there was no difference in the proportion of smolts between the control and melatonin-treated groups. Thus, melatonin feeding mimicked the effects of short photoperiod, which delays but does not completely suppress smoltification. These results indicate that the day length is transduced into changes in the duration of nocturnal elevation in plasma melatonin levels, and that artificial modification of the plasma melatonin pattern possibly delays the physiological processes of smoltification induced by long-day photoperiodic treatment.     

Seasonal differences in melatonin concentrations and heart rates during sleep in obese subjects in Japan

During the past several decades, obesity has been increasing globally. In Japan, obesity is defined by a BMI of 25 kg/m² or over; 28.6 % of men and 20.6 % of women are obese. Obese people have an increased incidence of developing cardiovascular, renal, and hormonal diseases and sleep disorders. Obese people also have shortened sleep durations. We investigated seasonal differences in melatonin concentrations, heart rates, and heart rate variability during sleep in obese subjects in Japan. Five obese (BMI, 32.0?±?4.9 kg/m²) and five non-obese (BMI, 23.2?±?2.9 kg/m²) men participated in this study in the summer and winter. Electrocardiograms were measured continuously overnight in a climatic chamber at 26 °C with a relative humidity of 50 %. Saliva samples for melatonin were collected at 2300 hours, 0200 hours, and 0600 hours. We found that melatonin concentrations during sleep in obese subjects were significantly lower than those in non-obese subjects in the winter. Heart rate during sleep in winter was significantly higher than that in summer in both obese and non-obese subjects. Heart rate variability was not significantly different in the summer and winter in both obese and non-obese subjects. Our results show that decreased nocturnal melatonin concentrations during winter in obese men may be related to higher heart rates, and this may suggest that obese men are at an increased risk of a cardiovascular incident during sleep, especially in the winter.     

Urinary melatonin rhythms during sleep deprivation in depressed patients and normals

Melatonin excretion was measured in 8 hour urine aliquots for eight healthy controls and six depressed patients. Both groups had similar diurnal rhythms, with increased melatonin excretion during the night. When subjects were sleep deprived, remaining awake and active in continuous light from 7 a.m. one morning until 11 p.m. the following day, the diurnal rhythm in melatonin excretion remained unchanged. These data in man appear to be inconsistent with previous studies in rats showing rapid light-induced suppression of the nocturnal rise in pineal melatonin synthesis.     

Evaluation of 5-hydroxytryptophan administration as a test of pineal function in humans

We tested the hypothesis that acute 5-hydroxytryptophan (5-HTP) administration would cause an increase in concentrations of plasma melatonin to levels observed during the spontaneous nocturnal melatonin surge. We administered 5-HTP orally (5-12 mg/kg) to 10 healthy children and 5 healthy adults, and measured melatonin concentrations in plasma samples obtained every 30 min for 3-6 h. There was no appreciable increase in melatonin after 5-HTP stimulation, even though a melatonin increase has been reported in sheep treated with 5-HTP.