Circadian Rhythm Of Melatonin Release Research Articles

Efficacy and Safety of Prolonged-Release Melatonin for Primary Insomnia in Elderly Patients

Objective: Insomnia, a prevalent sleep disorder affecting the elderly, necessitates effective and safe treatment options. This study explores the potential of the prolonged-release melatonin (Circadin<sup>®</sup>) addressing insomnia in elderly patients.Methods: The 8-week prospective observational study involved 115 participants, assessing sleep parameters using the Pittsburgh Sleep Quality Index (PSQI) and WHO-5 Well-being Index. The prolonged-release melatonin (2 mg) was administered nightly, and assessments were conducted at baseline, 4 weeks, and 8 weeks.Results: Both per protocol and last observation carried forward (LOCF) analyses consistently revealed significant improvements in sleep latency, total sleep time, sleep efficiency, and overall well-being. Notably, sleep latency decreased after 4 and 8 weeks, while total sleep time and sleep efficiency increased, reflecting longer and more restful sleep. The WHO-5 Well-being Index exhibited noticeable enhancement. Adverse events, including dizziness and heartburn, were manageable. Despite a high dropout rate, this drug’s potential as an efficacious and safe treatment option for elderly insomnia patients was evident, aligned with prior research.Conclusion: The prolonged-release formulation’s resemblance to the natural circadian rhythm of melatonin release offers advantages over conventional medications. This study contributes to understanding the prolonged-release melatonin’s promise as a valuable therapeutic alternative, encouraging further investigation into its longterm effects and optimal implementation.

Sleep, Melatonin, and the Menopausal Transition: What Are the Links?

The pineal hormone Melatonin plays an important role in the regulation of the circadian sleep/wake cycle, mood, and perhaps immune functions, carcinogensis and reproduction. The human circadian rhythm of melatonin release from the pineal gland is tightly synchronized with the habitual hours of sleep. Peri- and postmenopausal women often complain of difficulties initiating and/or maintaining sleep, with frequent nocturnal and early morning awakenings. In this review we discuss the pathophysiology of melatonin function as it relates to sleep disorders in menopausal women, highlighting the potential use of exogenous melatonin during the menopausal transition and beyond.

Altered melatonin secretion and circadian gene expression with increased proinflammatory cytokine expression in early-stage sepsis patients

Inflammatory and immune responses, as well as melatonin secretion, are affected by circadian regulation. Abnormal circadian rhythm of melatonin release has been reported to be associated with the later stages of sepsis; however, its role in the early stages of sepsis is unclear. We studied 11 septic and 11 non-septic patients in our intensive care unit (ICU). Peripheral blood was drawn at 4-h intervals on the first day, beginning at 2:00 p.m., over a total period of 24 h. Plasma levels of melatonin, tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) were measured by radioimmunoassay or enzyme-linked immunosorbent assay (ELISA). Messenger RNA levels of circadian genes Cry-1 and Per-2 were analyzed using quantitative real-time PCR. Results show the circadian rhythm of melatonin secretion was altered in the early stages of sepsis. The melatonin secretion acrophase occurred earlier in septic patients at 6:00 p.m., compared with at 2:00 a.m. in non-septic ICU patients. Compared with the non-septic group, both Cry-1 and Per-2 expression were significantly decreased while TNF-α and IL-6 expression were significantly increased in septic patients [TNF-α, 64.1 (43.6-89.1) vs. 11.4 (10.4-12.5) ng/ml; IL-6, 41.2 (35.7-50.8) vs. 19.1 (16-136.7) ng/ml; median (range), both P=0.04]. The peak concentrations of TNF-α and IL-6 were shown to be in concordance with the rhythm of melatonin secretion. The circadian rhythm of melatonin secretion and circadian gene expression were altered in the early stages of sepsis, which likely led to the changes in pro-inflammatory cytokine release. These findings shed light on the potential link between circadian rhythm and the progression of early-stage sepsis.

Differential Contribution of Rod and Cone Circadian Clocks in Driving Retinal Melatonin Rhythms in Xenopus

BackgroundAlthough an endogenous circadian clock located in the retinal photoreceptor layer governs various physiological events including melatonin rhythms in Xenopus laevis, it remains unknown which of the photoreceptors, rod and/or cone, is responsible for the circadian regulation of melatonin release.Methodology/Principal FindingsWe selectively disrupted circadian clock function in either the rod or cone photoreceptor cells by generating transgenic Xenopus tadpoles expressing a dominant-negative CLOCK (XCLΔQ) under the control of a rod or cone-specific promoter. Eyecup culture and continuous melatonin measurement revealed that circadian rhythms of melatonin release were abolished in a majority of the rod-specific XCLΔQ transgenic tadpoles, although the percentage of arrhythmia was lower than that of transgenic tadpole eyes expressing XCLΔQ in both rods and cones. In contrast, whereas a higher percentage of arrhythmia was observed in the eyes of the cone-specific XCLΔQ transgenic tadpoles compare to wild-type counterparts, the rate was significantly lower than in rod-specific transgenics. The levels of the transgene expression were comparable between these two different types of transgenics. In addition, the average overall melatonin levels were not changed in the arrhythmic eyes, suggesting that CLOCK does not affect absolute levels of melatonin, only its temporal expression pattern.Conclusions/SignificanceThese results suggest that although the Xenopus retina is made up of approximately equal numbers of rods and cones, the circadian clocks in the rod cells play a dominant role in driving circadian melatonin rhythmicity in the Xenopus retina, although some contribution of the clock in cone cells cannot be excluded.

Lack of circadian regulation of in vitro melatonin release from the pineal organ of salmonid teleosts

In many teleost species, the photoreceptive pineal organ harbors the circadian clock that regulates melatonin release in the pineal organ itself. However, the pineal organ of three salmonids (rainbow trout Oncorhynchus mykiss, masu salmon Oncorhynchus masou, and sockeye salmon Oncorhynchus nerka) did not exhibit circadian rhythms in melatonin release when maintained under constant darkness (DD) in vitro, suggesting that the pineal organs of all salmonids lack the circadian regulation of melatonin production. To test this hypothesis, the pineal organ of seven salmonids (common whitefish Coregonus lavaretus, grayling Thymallus thymallus, Japanese huchen Hucho perryi, Japanese charr Salvelius leucomaenis pluvius, brook trout Salvelius fontinalis, brown trout Salmo trutta and chum salmon Oncorhynchus keta) and closely related osmerids (ayu Plecoglossus altivelis altivelis and Japanese smelt Hypomesus nipponensis) were individually maintained in flow-through culture at 15 °C under several light conditions. Under light–dark cycles, the pineal organ of all species showed a rhythmic melatonin release with high rates during the dark phase. Under DD, the osmerid pineal organs exhibited circadian rhythms in melatonin release with high rates only during the subjective-night but the salmonid pineal organs constantly released melatonin at high rates. Under constant light, melatonin release was suppressed in all species. The pineal organ of rainbow trout maintained at different temperature (15, 20 or 25 °C) under DD released melatonin with high rates but the amount of melatonin released was temperature-sensitive (highest at 20 °C). Thus, melatonin release from the pineal organ of osmerids is regulated by both light and circadian clock but the circadian regulation is lacking in salmonids. These results indicate that ancestral salmonids lost the circadian regulation of melatonin production after the divergence from osmerid teleosts.

Light, dark, and melatonin: emerging evidence for the importance of melatonin in ocular physiology

Melatonin is a hormone, which is mainly produced by the pineal gland, a vestigial eye. Rather than the rods and cones, it is a newly discovered subgroup of photosensitive retinal ganglion cells, which is responsible for mediating the light-dark cycles, thus regulating melatonin's secretion. One of the correlates of the circadian rhythm of melatonin release is the habitual sleep pattern. Patients with circadian rhythm sleep disorders, including some blind patients with no light-induced suppression of melatonin, benefit from melatonin treatment. Melatonin is synthesized in the retina, lens, ciliary body as well as other parts of the body. In this review, we discuss the physiological roles of melatonin in the eye, as well as the potential therapeutic avenues currently under study.

Features of the Circadian Clock in the Avian Pineal Gland

The effects of different environmental factors on the circadian rhythm of melatonin release by chicken pineal glands were analyzed in 5-day experiments in vitro. Periodic, brief 4-h illumination at around midnight resulted in phase inversion in 2 days. Exposure to repeated 41 degrees C temperature elevation at around midnight caused no phase shift in the basic melatonin rhythm. Repeated inversion of the polarity of the magnetic field also affected rhythmic melatonin release. Additionally, expressions of Per1, Per2, Per3, Bmal1, Bmal2, Clock, Cry1, and Cry2 clock genes were simultaneously detected in single chicken pineal glands.

Signal transmission from the suprachiasmatic nucleus to the pineal gland via the paraventricular nucleus: analysed from arg-vasopressin peptide, rPer2 mRNA and AVP mRNA changes and pineal AA-NAT mRNA after the melatonin injection during light and dark periods

Arg-vasopressin (AVP) containing neurons are one of the output paths from the suprachiasmatic nucleus (SCN), the center of the biological clock. AVP mRNA transcription is controlled by a negative feedback loop of clock genes. Circadian rhythm of melatonin release from the pineal gland is regulated by the SCN via the paraventricular nucleus (PVN). To clarify the transduction system of circadian signals from the SCN to the pineal gland, we determined the effects of melatonin injection (1 mg/kg, i.p.) during light and dark periods on Per2 and AVP mRNAs in the SCN and PVN, in addition to arylalkylamine N-acetyltransferase ( AA- NAT) and inducible cAMP early repressor ( ICER) mRNAs in the pineal gland of rats using RT-PCR. AVP peptide contents were also measured in the SCN and PVN. AVP content in the SCN decreased during the light period, while no changes were observed in the PVN. In the SCN, Per2 mRNA increased during both light and dark periods. In the PVN, Per2 decreased during the light period and increased during the dark period at 180 min after melatonin injection. In the pineal gland, Per2 mRNA increased between 60 and 180 min after the melatonin injection during the light period, while it did not significantly change during the dark period. The AA- NAT mRNA varied similar to the Per2 mRNA changes. These results might suggest that the different responses to melatonin in the pineal gland during the light and dark periods was originated in the changes of Per2 in the PVN via SCN.

Circadian rhythm of melatonin release from the photoreceptive pineal organ of a teleost, ayu (Plecoglossus altivelis) in flow-thorough culture.

In the present study, we tested whether the pineal organ of ayu (Plecoglossus altivelis), an osmerid teleost close relative of salmonids, harbours a circadian oscillator regulating rhythmic melatonin release using flow-through culture. The pineal organ maintained under light/dark cycles released melatonin in a rhythmic fashion with high levels during the dark phase. A circadian rhythm of melatonin release persisted in constant darkness for at least four cycles. Characteristics of the circadian rhythm (free-running period, phase and amplitude) exhibited small variations among cultures when the data was normalized, indicating that this system is sufficient for the analysis of the circadian rhythm both at qualitative and quantitative levels. Six-hour extension of the light phase from the normal onset time of the dark phase or exposure to constant light for 36 or 48 h before transfer to constant darkness significantly inhibited melatonin release. Phase shifts in the circadian rhythm of melatonin release were also observed. Thus, the ayu pineal organ contains all the three essential components of the circadian system (a circadian clock, the photoreceptor responsible for photic entrainment of the clock, and melatonin generating system as an output pathway). This system should provide a useful model for analysing the physiological and molecular basis of the vertebrate circadian system. In addition, further comparative studies using salmonids and related species including ayu will provide some insight into the evolution of the roles of the pineal organ in the vertebrate circadian system.

Are Changes in MAPK/ERK Necessary or Sufficient for Entrainment in Chick Pineal Cells?

Chick pineal cells in culture display a circadian rhythm of melatonin release. Light pulses can entrain (phase shift) the rhythm. One candidate for the photoentrainment pathway uses a mitogen-activated protein kinase (MAPK), also known as extracellular signal-regulated kinase (ERK). We tested the hypothesis that activation of ERK (by phosphorylation to p-ERK) is necessary and/or sufficient for entrainment by measuring the ability of several drugs, light, and other perturbations to change levels of p-ERK and to induce phase shifts in the melatonin rhythm. If changes in the levels of p-ERK are sufficient for photoentrainment, then all perturbations that reduce its level must induce light-like phase shifts, and all those that increase its level must induce dark-like phase shifts. If such changes are necessary for photoentrainment, then light pulses must reduce p-ERK levels, and the duration of the light pulse, the magnitude and duration of the change in p-ERK, and the size of the phase shift must correlate. We found five perturbations that reduced p-ERK levels. Of these, two induced light-like phase shifts (PD 98059 and caffeine), one induced dark-like phase shifts (SB203580), and two did not induce phase shifts at all (U0126 and omitting a medium change). Serum increased p-ERK levels without inducing any phase shifts. Finally, light pulses did not elicit changes in p-ERK, nor was there a diurnal rhythm in p-ERK levels, nor could rapid changes in p-ERK levels have accounted for duration effects of light pulses on phase shifts. Taken together, these results argue strongly against the hypothesis that reduction (or increases) in MAPK/ERK activation is necessary or sufficient for entrainment in chick pineal cells.

In vitro photic entrainment of the circadian rhythm in melatonin release from the pineal organ of a teleost, ayu ( Plecoglossus altivelis) in flow-through culture

The effects of light on the circadian rhythm in melatonin release from the pineal organ of a teleost, ayu ( Plecoglossus altivelis) were investigated in flow-through culture. Under the reversed light–dark (LD) cycle, the melatonin rhythm phase shifted as compared with those under the normal LD cycle. This phase shift persisted even under constant darkness (DD). Single 6-h light pulses starting at six different circadian phases under DD acutely suppressed melatonin release. Phase-dependent phase shifts in the circadian rhythm of melatonin release were also observed. The phase response curve to light pulses in the ayu pineal organ is typical of that found in many circadian systems. Thus, the ayu pineal organ should provide a useful model for analyzing the physiological and molecular basis of the entrainment mechanism of vertebrate circadian system.

Photoperiod affects amplitude but not duration of in vitro melatonin production in the ruin lizard (Podarcis sicula).

The pineal gland and its major output signal melatonin have been demonstrated to play a central role in the seasonal organization of the ruin lizard Podarcis sicula. Seasonal variations in the amplitude of the nocturnal melatonin signal, with high values in spring as compared to low values in summer and autumn, have been found in vivo. The authors examined whether the pineal gland of the ruin lizard contains autonomous circadian oscillators controlling melatonin synthesis and whether previously described seasonal variations of in vivo melatonin production can also be found in isolated cultured pineal glands obtained from ruin lizards in summer and winter. In vitro melatonin release from isolated pineal glands of the ruin lizard persisted for 4 days in constant conditions. Cultured explanted pineal glands obtained from animals in winter and summer showed similar circadian rhythms of melatonin release, characterized by damping of the amplitude of the melatonin rhythm. Although different photoperiodic conditions were imposed on ruin lizards before explantation of pineal glands, the authors did not find any indication for corresponding differences in the duration of elevated melatonin in vitro. Differences were found in the amplitude of in vitro melatonin production in light/dark conditions and, to a lesser degree, in constant conditions. The presence of a circadian melatonin rhythm in vitro in winter, although such a rhythm is absent in vivo in winter, suggests that pineal melatonin production is influenced by an extrapineal oscillator in the intact animal that may either positively or negatively modulate melatonin production in summer and winter, respectively.

Microarray technology in studying the effect of melatonin on gene expression in the mouse heart.

In recent years, numerous and sometimes contradictory data on the key role of the epiphysis (pineal gland) in aging and the antioxidant processes have attracted much attention [1, 2]. In various animal species, the night production of the epiphyseal indole hormone melatonin decreases with age, and the circadian rhythm of melatonin release into circulation is disturbed [2, 3]. After epiphysectomia, the animal’s life span decreases, whereas administration of melatonin to animals of various species (mice, rats, Drosophila, and flatworms) increases their life span [2, 4]. We recently showed that long-term melatonin administration to female CBA mice resulted in a retarded dynamics of the biologicalage markers and increased the life span [5]. However, little is known about the molecular mechanisms underlying the melatonin effect on gene expression, though the hormone was found to be mainly accumulated in the cell nuclei and incorporated into chromatin in most tissues and organs [6]. We studied the effect of melatonin on the expression of genes in the heart of CBA mice after the hormone was administered at a geroprotective dose.

Circadian rhythm of melatonin release in pineal gland culture: arg-vasopressin inhibits melatonin release

The mammalian pineal gland is known to receive a noradrenergic sympathetic efferent signal from the suprachiasmatic nucleus (SCN) via the superior cervical ganglion. Arg-vasopressin (AVP) containing neurons in the SCN is one of the output paths of circadian information to the other brain areas. AVP release from the SCN is suppressed by melatonin. In turn, we determined the direct effect of AVP on melatonin release using pineal gland explant culture. AVP (1 μM) suppressed melatonin release. Noradrenaline stimulated melatonin release was attenuated by AVP. In turn, the expression of the melatonin synthesis enzyme arylalkylamine N-acetyltransferase mRNA in the rat SCN was reported. We measured melatonin content in the SCN in rats kept under the light–dark cycle and constant dim light. Melatonin in the SCN was higher during the dark period than that in the light. A similar tendency was also observed in the SCN of animals kept under a constant dim light. It was suggested that the reciprocal regulation of melatonin release and AVP release occurs in the SCN and pineal gland.

Photoperiodic information acquired and stored in vivo is retained in vitro by a circadian oscillator, the avian pineal gland.

Endogenous circadian rhythms have been described in a wide range of organisms from prokaryotes to man. Although basic circadian mechanisms at the molecular level are genetically fixed, certain properties of circadian rhythms at the organismic level can be modified by environmental conditions and subsequently retained for some time, even in organisms shielded from 24-hr environmental variations. To investigate the capacity of animals to acquire and store photoperiodic information, we examined activity and melatonin rhythms in house sparrows during synchronization to two different photoperiods and during subsequent prolonged darkness. Under constant environmental conditions, intact animals continued to have long feeding activity times when previously exposed to long days and short feeding activity times when previously exposed to short days. Correspondingly, significantly different durations of elevated melatonin in the plasma directly reflected the differences in night length during synchronization as well as during prolonged darkness. Additionally, we found a significant difference in the amplitude of the nocturnal melatonin signal, which also was conserved in prolonged darkness. To investigate whether the photoperiodic experience of an intact animal can be "memorized" by an isolated component of its circadian pacemaking system, we have investigated in vitro melatonin release during continuous darkness from explanted pineal glands of house sparrows after in vivo synchronization to two distinct photoperiods. Differences in the durations of elevated melatonin occurred during the first two cycles in culture and a difference in melatonin amplitude was detectable during the first night in culture. Our data indicate that photoperiodic patterns imposed on sparrows during in vivo synchronization can be maintained as an internal representation of time within the isolated pineal gland. Hence, the pineal gland, as one of the most significant components of the songbird circadian pacemaker, not only has the capacity to autonomously produce circadian rhythms of melatonin release but also is capable of storing biologically meaningful information experienced during previous cycles.

Circadian Rhythm-Dependent Development of Melatonin Effects and Tolerance to PHNO in Rats

Male Sprague–Dawley rats were given 12 days of continuous infusions of (+)-4-propyl-9-hyroxynapthoxazine (PHNO, 5μg/h), a highly selective dopamine D 2 receptor agonist, via subcutaneous ALZET ® osmotic pumps. Motor stimulant effects (locomotion and rearing) were monitored throughout the treatment period, including after the animals were injected with 2-iodo-melatonin (0.5 mg/kg) on days 8–10 and 13 after initiation of PHNO infusions. The rats (maintained on 12 L:12 D cycle) developed tolerance to the motor stimulant effects of PHNO during the day, and behavioral sensitization to PHNO during the night. Arousing rats with a vehicle injection transiently blocked the daytime tolerance. A more sustained environmental noise without handling of animals, which had a stronger effect on increasing motor activity of control rats, reversed tolerance to sensitization. Therefore, graded levels of arousal produce graded increases in motor activity in rats otherwise tolerant to the effects of PHNO. Daytime tolerance to PHNO was reversed to sensitization by 2-iodo-melatonin. This effect was more than an additive effect of drug + injection procedure stress. The differential development of nocturnal sensitization and diurnal tolerance to PHNO effects on motor activity may depend upon circadian rhythms in melatonin release, as well as on state of arousal.

Do entrainment conditions under light-dark cycles affect circadian rhythms in melatonin release from the photosensitive pineal organ of a teleost, ayu (Plecoglossus altivelis)?

Do entrainment conditions under light-dark cycles affect circadian rhythms in melatonin release from the photosensitive pineal organ of a teleost, ayu (Plecoglossus altivelis)?

Circadian rhythm of melatonin release in pineal gland organ culture: vasopressin suppresses melatonin release in rats

Circadian rhythm of melatonin release in pineal gland organ culture: vasopressin suppresses melatonin release in rats

Individual pineal cells in chick possess photoreceptive, circadian clock and melatonin-synthesizing capacities in vitro

Chick pineal cells express a circadian rhythm of melatonin release under light–dark (LD) cycles, with an increase during the dark period and a decrease during the light period, and this rhythm persists under constant darkness (DD). We cultured individual single pineal cells with 15 μl of medium per well in a Terasaki plate and measured melatonin secretion every 12 h under LD, DL and DD. Individual cells secreted more melatonin during the dark period than during the light period under both LD and DL conditions, and those rhythmic secretions persisted under DD. These results suggest that individual pineal cells in chick have photoreceptive, circadian clock and melatonin-synthesizing capacities.

A cationic channel regulated by a vertebrate intrinsic circadian oscillator.

Secretory cells of the chicken pineal gland exhibit light-sensitive circadian rhythms in melatonin release that persist in vitro. Melatonin secretion is positively regulated by cyclic AMP and intracellular Ca2+ (refs 8, 10-15). Cyclic AMP analogues are more effective at stimulating melatonin secretion during the circadian night owing in part to increased Ca2+ influx at those times. However, this cannot be attributed to increased activity of L-type Ca2+ channels. Here we describe an unusual 40-pS cationic channel (ILOT) in cultured chicken pineal cells that is permeable to Ca2+ and active in the night but not during the day. ILOT is not voltage- or stretch-activated, it has a characteristically long open time, and its gating persists in excised inside-out patches in the absence of Ca2+ or cyclic nucleotides. Daily rhythms in ILOT gating are also observed in previously entrained chicken pineal cells free-running under constant dark conditions. Nighttime ILOT activity is not suppressed by brief light pulses.