Regulation Of Melatonin Secretion Research Articles

Methodology for the optimization of artificial lighting considering both visual and non-visual effects

The effect of light on human physiology as well as its non-image forming effects have been known for several years. An important milestone in understanding the non-visual effects of light was the discovery of a new type of photoreceptor namely the intrinsically photosensitive retinal ganglion cells, or ipRGCs which play a vital role in the human circadian system. The non-visual effects of light are the following: regulation of melatonin secretion, circadian entrainment and modification of body temperature. With the advent of solid-state lighting, it is possible to precisely regulate the spectral power distribution of artificial lighting, so as to favour the human circadian rhythm. The scope of this paper is to present a conceptual methodology for the evaluation of artificial lighting systems with regards to visual, circadian effects and their energy consumption. In other words, this paper aims to outline an assessment process for lighting designers by elaborating not only on the visual aspects of each lighting system but also the melanopic effects and its energy efficiency.

CircRNA-WNK2 Acts as a ceRNA for miR-328a-3p to Promote AANAT Expression in the Male Rat Pineal Gland.

Noncoding RNAs (ncRNAs), including microRNAs (miRNAs) and circular RNAs (circRNAs), which are expressed with a daily rhythm in the rat pineal gland, are associated with the regulation of melatonin secretion and other biological functions. However, the mechanisms of these molecules in the rat pineal gland are not yet fully understood. In this study, we found that circR-WNK2 was highly expressed at night, which may be involved in the regulation of melatonin secretion through the competitive endogenous RNA (ceRNA) mechanism. By dual luciferase reporter, RNA pull-down, and fluorescence in situ hybridization (FISH) assays, we found that miR-328a-3p can target circR-WNK2 and the Aa-nat mRNA 3'UTR. Transfection experiments indicated that circR-WNK2 could competitively bind to miR-328a-3p, reduce miR-328a-3p expression, and promote Aa-nat gene expression and melatonin secretion. And by constructing a superior cervical ganglionectomy (SCGx) rat model, we found that ncRNAs expression in the pineal gland was regulated by signals from the suprachiasmatic nucleus. This finding supports the hypothesis that these noncoding RNAs may interact to shape the circadian rhythm through transcriptional processing in melatonin synthesis.

Pinopsin and photoreception in the pineal organ of the domestic turkey during post-hatching development

The avian pineal organ is photosensitive because of the presence of photopigments, of which pinopsin seems to be one of the most important. This organ is subject to far-reaching changes during post-hatching development, but evidence regarding pinopsin presence and direct photoreception during this time is lacking. This study was carried out to demonstrate the following: 1) the structures showing immunoreactivity to pinopsin in the turkey pineal organ, 2) the changes of these structures during development, 3) the pinopsin localization in pinealocytes in monolayer cultures, and 4) the role of direct photoreception in the regulation of melatonin secretion in pineal organs in adult turkeys. Pinopsin immunoreactivity was localized in the apical extensions of columnar cells limiting the follicular lumen, in fiber-like structures located between columnar cells in the inner part of follicle wall, in string-shapes or small spherical structures distributed in the outer part of follicle wall and in amorphous material inside the follicle lumen. In young birds, immunoreactivity was also sporadically noted in cell bodies of rudimentary receptor pinealocytes. The distribution of pinopsin showed prominent age-dependent changes, including a subsequent increase in pinopsin-positive structures in the outer part of the follicle wall and a prominent reduction in the number and size of positive apical extensions in 40- and 56-week-old turkeys. These data demonstrate that the role of secretory pinealocytes in pineal photoreception increases with age. In monolayer cultures, all pinealocytes showed strong reactions in club- or bulbous-shaped prolongations. The pineal organs of adult birds were less sensitive to light exposition at night than those of young turkeys, which points to differences in light sensitivity between rudimentary receptor and secretory pinealocytes. However, direct photoreception could play an important role in the regulation of melatonin secretion in adult turkeys.

Roles of Direct Photoreception and the Internal Circadian Oscillator in the Regulation of Melatonin Secretion in the Pineal Organ of the Domestic Turkey: A Novel In Vitro Clock and Calendar Model.

The regulation of melatonin secretion in the avian pineal organ is highly complex and shows prominent interspecies differences. The aim of this study was to determine the roles of direct photoreception and the internal oscillator in the regulation of melatonin secretion in the pineal organ of the domestic turkey. The pineal organs were collected from 12-, 13- and 14-week-old female turkeys reared under a 12 L:12 D cycle with the photophase from 07.00 to 19.00, and were incubated in superfusion culture for 3–6 days. The cultures were subjected to different light conditions including 12 L:12 D cycles with photophases between 07.00 and 19.00, 13.00 and 01.00 or 01.00 and 13.00, a reversed cycle 12 D:12 L, cycles with long (16 L:8 D) and short (8 L:16 D) photophases, and continuous darkness or illumination. The pineal organs were also exposed to light pulses of variable duration during incubation in darkness or to periods of darkness during the photophase. The secretion of melatonin was determined by direct radioimmunoassay. The turkey pineal organs secreted melatonin in a well-entrained diurnal rhythm with a very high amplitude. Direct photoreception as an independently acting mechanism was able to ensure quick and precise adaptation of the melatonin secretion rhythm to changes in light-dark conditions. The pineal organs secreted melatonin in circadian rhythms during incubation in continuous darkness or illumination. The endogenous oscillator of turkey pinealocytes was able to acquire and store information about the light-dark cycle and then to generate the circadian rhythm of melatonin secretion in continuous darkness according to the stored data. The obtained data suggest that the turkey pineal gland is highly autonomous in the generation and regulation of the melatonin secretion rhythm. They also demonstrate that the turkey pineal organ in superfusion culture is a valuable model for chronobiological studies, providing a highly precise clock and calendar. This system has several features which make it an attractive alternative to other avian pineal glands for circadian studies.

TMEM16A and TMEM16B channel proteins generate Ca2+-activated Cl− current and regulate melatonin secretion in rat pineal glands

Pinealocytes regulate circadian rhythm by synthesizing and secreting melatonin. These cells generate action potentials; however, the contribution of specific ion channels to melatonin secretion from pinealocytes remains unclear. In this study, the involvement and molecular identity of Ca2+-activated Cl- (ClCa) channels in the regulation of melatonin secretion were examined in rat pineal glands. Treatment with the ClCa channel blockers, niflumic acid or T16Ainh-A01, significantly reduced melatonin secretion in pineal glands. After pineal K+ currents were totally blocked under whole-cell patch clamp conditions, depolarization and subsequent repolarization induced a slowly activating outward current and a substantial inward tail current, respectively. Both of these current changes were dependent on intracellular Ca2+ concentration and inhibited by niflumic acid and T16Ainh-A01. Quantitative real-time PCR, Western blotting, and immunocytochemical analyses revealed that TMEM16A and TMEM16B were highly expressed in pineal glands. siRNA knockdown of TMEM16A and/or TMEM16B showed that both channels contribute to ClCa currents in pinealocytes. Conversely, co-expression of TMEM16A and TMEM16B channels or the expression of this tandem channel in HEK293 cells mimicked the electrophysiological characteristics of ClCa currents in pinealocytes. Moreover, bimolecular fluorescence complementation, FRET, and co-immunoprecipitation experiments suggested that TMEM16A and TMEM16B can form heteromeric channels, as well as homomeric channels. In conclusion, pineal ClCa channels are composed of TMEM16A and TMEM16B subunits, and these fluxes regulate melatonin secretion in pineal glands.

Adrenergic control of pinealocyte chondriome - an in vitro study.

Norepinephrine released from sympathetic innervation plays the main role in the regulation of melatonin secretion in mammalian pinealocytes. The present study was conducted for the following reasons: 1) to establish whether the pinealocyte chondriome is controlled by norepinephrine, 2) to determine the effect of adrenergic stimulation on mitochondria, and 3) to characterize adrenoceptors involved in the regulation of the chondriome. The static organ culture of the pineal gland was used. The explants were incubated for 5 consecutive days in control medium and between 20:00 and 08:00 in medium with the presence of 10 μM norepinephrine - adrenergic agonist; isoproterenol - beta-adrenoceptor agonist; cirazoline, methoxamine, M-6364 - alfa1 - adrenoceptors agonists or PMA - activator of PKC. The explants were then subjected to ultrastructural examination and morphometric analysis. The incubation of explants in the presence of norepinephrine or isoproterenol caused a decrease in the relative volume and the numerical density of mitochondria and induced an increase in the percentage of free mitochondria in pinealocytes. Significant changes in these parameters were not observed after treatment with methoxamine, cirazoline, M-6463 and PMA. The results obtained show that the chondriome of pig pinealocytes is controlled by norepinephrine acting via beta-adrenoceptors. Adrenergic stimulation, repeated for five consecutive days of organ culture, causes a decrease in the number of mitochondria and a shift in the distribution of mitochondria from the form of networks and filaments into the form of single particles. This indicates the intensive remodeling of the mitochondria network, which is closely linked to the metabolic status of the cell.

In the Heat of the Night: Thermo-TRPV Channels in the Salmonid Pineal Photoreceptors and Modulation of Melatonin Secretion.

Photoperiod plays an essential role in the synchronization of metabolism, physiology, and behavior to the cyclic variations of the environment. In vertebrates, information is relayed by the pineal cells and translated into the nocturnal production of melatonin. The duration of this signal corresponds to the duration of the night. In fish, the pinealocytes are true photoreceptors in which the amplitude of the nocturnal surge is modulated by temperature in a species-dependent manner. Thus, the daily and annual variations in the amplitude and duration of the nocturnal melatonin signal provide information on daily and calendar time. Both light and temperature act on the activity of the penultimate enzyme in the melatonin biosynthesis pathway, the arylalkylamine N-acetyltransferase (serotonin → N-acetylserotonin). Although the mechanisms of the light/dark regulation of melatonin secretion are quite well understood, those of temperature remain unelucidated. More generally, the mechanisms of thermoreception are unknown in ectotherms. Here we provide the first evidence that two thermotransient receptor potential (TRP) channels, TRPV1 and TRPV4, are expressed in the pineal photoreceptor cells of a teleost fish, in which they modulate melatonin secretion in vitro. The effects are temperature dependent, at least for TRPV1. Our data support the idea that the pineal of fish is involved in thermoregulation and that the pineal photoreceptors are also thermoreceptors. In other nervous and nonnervous tissues, TRPV1 and TRPV4 display a ubiquitous but quantitatively variable distribution. These results are a fundamental step in the elucidation of the mechanisms of temperature transduction in fish.

Regulation of melatonin secretion in the pineal organ of the domestic duck--an in vitro study.

The aim of study was to determine the mechanisms regulating melatonin secretion in the pineal organs of 1-day-old and 9-month-old domestic ducks. The pineals were cultured in a superfusion system under different light conditions. Additionally, some explants were treated with norepinephrine. The pineal glands of 1-day-old ducks released melatonin in a well-entrained, regular rhythm during incubation under a 12 hrs light:12 hrs dark cycle and adjusted their secretory activity to a reversed 12 hrs dark:12 hrs light cycle within 2 days. In contrast, the diurnal changes in melatonin secretion from the pineals of 9-month-old ducks were largely irregular and the adaptation to a reversed cycle lasted 3 days. The pineal organs of nestling and adult ducks incubated in a continuous light or darkness secreted melatonin in a circadian rhythm. The treatment with norepinephrine during photophases of a light-dark cycle resulted in: 1) a precise adjustment of melatonin secretion rhythm to the presence of this catecholamine in the culture medium, 2) a very high amplitude of the rhythm, 3) a rapid adaptation of the pineal secretory activity to a reversed light-dark cycle. The effects of norepinephrine were similar in the pineal organs of nestlings and adults. In conclusion, melatonin secretion in the duck pineal organ is controlled by three main mechanisms: the direct photoreception, the endogenous generator and the noradrenergic transmission. The efficiency of intra-pineal, photosensitivity-based regulatory mechanism is markedly lower in adult than in nestling individuals.

Adrenergic activation of melatonin secretion in ovine pineal explants in short-term superfusion culture occurs via protein synthesis independent and dependent phenomena.

The ovine pineal is generally considered as an interesting model for the study on adrenergic regulation of melatonin secretion due to some functional similarities with this gland in the human. The present investigations, performed in the superfusion culture of pineal explants, demonstrated that the norepinephrine-induced elevation of melatonin secretion in ovine pinealocytes comprised of two subsequent periods: a rapid increase phase and a slow increase phase. The first one included the quick rise in release of N-acetylserotonin and melatonin, occurring parallel to elevation of NE concentration in the medium surrounding explants. This rapid increase phase was not affected by inhibition of translation. The second, slow increase phase began after NE level had reached the maximum concentration in the culture medium and lasted about two hours. It was completely abolished by the treatment with translation inhibitors. The obtained results showed for the first time that the regulation of N-acetylserotonin synthesis in pinealocytes of some species like the sheep involves the on/off mechanism, which is completely independent of protein synthesis and works very fast. They provided strong evidence pointing to the need of revision of the current opinion that arylalkylamines N-acetyltransferase activity in pinealocytes is controlled exclusively by changes in enzyme abundance.

Lack of effect of ghrelin treatment on melatonin production in rat pineal and Harderian glands

The effects of ghrelin, a peptide hormone secreted from the stomach, on melatonin remain unknown. The aim of the study was to investigate possible ghrelin-melatonin interactions by studying the effect of ghrelin treatment on melatonin production in rat pineal and Harderian glands. Young (9 weeks) and old (20 months) male Wistar rats, maintained under a light:dark cycle regimen of 12:12, were assigned randomly to either a single subcutaneous (s.c.) injection of saline or ghrelin (1 μg/rat or 15 μg/rat) 1 h before sacrifice in the middle of the dark phase, or repeated s.c. saline or ghrelin injections (15 μg/rat), 3, 2 and 1 h before sacrificed in the middle of the dark phase. Neither ghrelin doses (1 μg/rat or 15 μg/rat) nor type of treatment (acute or repeated) influenced melatonin levels or the melatonin synthesizing enzymes N-acetyltransferase and hydroxyindole- O-methyltransferase activities, either in pineal gland or in Harderian glands. At the concentrations used, ghrelin does not influence melatonin production in rat pineal and Harderian glands, and therefore is not involved in the regulation of melatonin secretion, at least under our experimental conditions.

Pharmacological characterisation of oxytocin binding sites in the ovine pineal gland

Both oxytocin (OT) and [Arg 8]vasopressin (AVP) are found within the ovine pineal gland and may function to modulate melatonin secretion. However, the receptors which mediate the actions of these peptides have yet to be characterised. Preliminary studies of ovine pineal microsomal cell membranes showed binding of [ 3H]OT (79±9 fmol/mg) 10 times greater than binding of [ 3H]AVP (8±3 fmol/mg). Saturation studies using either [ 3H]OT or the selective OT receptor ligand [ 125I]d(CH 2) 5[Tyr(Me) 2,Thr 4,Orn 8,Tyr-NH 2 9]-vasotocin (OTA) revealed high affinity, single site kinetics ( K d=1.72±0.32 nM; B max=68±18 fmol/mg). Binding of [ 3H]AVP was more effectively displaced by OT than AVP, suggesting that binding may be due to cross-reaction with the OT binding site. Displacement of [ 3H]OT using a range of selective agonists and antagonist analogues revealed pharmacological characteristics similar to [ 3H]OT binding sites in the ovine and rat uterus. These data show that the ovine pineal expresses a high density of OT binding sites which may participate in the regulation of melatonin secretion.

Immunohistochemical Localization of D-Aspartate in the Rat Pineal Gland

Specific polyclonal antibody was raised against D-aspartate (D-Asp) which had been conjugated to glutaraldehyde and was purified by affinity chromatography. Immunohistochemical staining of rat pineal gland with the antibody demonstrated the presence of D-Asp in the cytoplasm of pinealocytes, the predominant cell type in this gland. D-Asp immunoreactivity was more evident in the distal region than in the proximal region of the gland. Pinealocytes in the distal region are presumably involved in the synthesis and secretion of the pineal hormone, melatonin, and the results of staining may indicate some yet unknown role of D-Asp in the regulation of melatonin secretion.

Elevated Nighttime Activity of Chick Pineal / Channels Requires Protein Synthesis

Free-running circadian rhythms in melatonin secretion persist in dissociated chick pineal cells. Calcium and cyclic AMP interact at several levels in the regulation of melatonin biosynthesis and secretion. Extracellular Ca2+ is required for optimal stimulation of melatonin secretion by cAMP analogues and protagonists. Increased Ca2+ influx during the circadian night is thought to play a role in the circadian clock regulation of melatonin secretion. We have recently described a nonselective cationic channel, ILOT, in chick pineal cells that is regulated by the intrinsic circadian oscillator. Active ILOT channels are detected only during the nighttime and may explain the nocturnal increase in Ca2+ influx. The mechanism by which the activity of ILOT is regulated by the circadian oscillator is not known. In the present study, the effect of the translational inhibitor anisomycin (10(-6) M) on the nighttime activity of ILOT channels was examined. The results show that protein synthesis is required for the detection of ILOT channel activity during the nighttime in cells maintained on light-dark cycles or constant dark conditions.

Regulation of melatonin secretion by light in the isolated pineal organ of the white sucker (Catostomus commersoni).

The effects of different lighting conditions and physical parameters of light were investigated in the isolated pineal organ of the white sucker kept under static or superfusion culture. The secretion of pineal melatonin is directly controlled by the photoperiod and completely suppressed under constant illumination. When pineal organs are exposed to unexpected light at night, the secretion of melatonin is significantly reduced within 15 min and reaches basal value after 30-35 min of light treatment. The inhibition of melatonin secretion by unexpected light at night depends on the irradiance, duration, timing of the treatment and the lighting history of the pineal organ.

Melatonin Response to Atrial Natriuretic Peptide Administration in Healthy Volunteers

Recent observations have demonstrated that the pineal hormone melatonin (MLT) plays a role in the neuroendocrine control of the cardiovascular system. On the other hand, it has been observed that the cardiac hormone alpha-atrial natriuretic peptide (ANP) may regulate the neuroendocrine functions. The present study was carried out to investigate the possible relationship between cardiac and pineal endocrine functions. Six healthy volunteers were treated on two different occasions with placebo or ANP at a dose of 0.1 mg i.v. as a bolus. An increase of greater than 100% in MLT serum levels was seen in 2/6 subjects. These preliminary results would suggest that ANP may play a role in the regulation of MLT secretion. Further studies will be needed to define better the cardiac-pineal interactions.

Regulation of melatonin secretion in a photoreceptive pineal organ: an in vitro study in the pike

The pineal organ (or pineal) of a teleost fish, the pike, contains typical (cone-like) and modified photoreceptor cells. Both are involved in indole metabolism, including melatonin production. How photoperiod controls melatonin biosynthesis in organs containing mainly photoreceptor cells, remains unclarified. To tackle this question we have used cultured pike pineal organs to investigate the variations in (1) the activity of the arylalkylamine-N-acetyltransferase (NAT), which is involved in the biosynthesis of melatonin, in static culture and (2) melatonin release in a perifusion system. Serum melatonin was also quantified in pike kept under a 24 hr light/dark (LD) cycle. Under LD conditions, NAT activity, melatonin release, and serum melatonin levels were high during the scotophase and low during the photophase. High-amplitude rhythms in NAT activity and melatonin release were maintained during three 24 hr cycles in constant darkness and a low-amplitude rhythm of NAT persisted in constant illumination. Midnight illumination induced a dramatic decrease of NAT activity and melatonin release. Darkness at midday did not induce a rise of the melatonin release, which occurred only at the onset of the subjective scotophase. From the present data it is strongly suggested that the pineal of the pike contains a circadian oscillator--synchronized by the photoperiod--which generates the rhythms of NAT activity and of melatonin release. Melatonin release, which reflects the rhythmic activity of NAT, might largely contribute to the melatonin circulating levels. The circadian oscillations observed under constant conditions suggest that the oscillator might behave differently in the pike, compared with intrapineal oscillators of 2 other species (lizard and chicken) under investigation.(ABSTRACT TRUNCATED AT 250 WORDS)

Evidence for a functional role of alpha-1 adrenoceptors in the regulation of melatonin secretion in man

Nocturnal plasma melatonin concentrations were reduced following administration of the α 1 adrenoceptor antagonist, prazosin (2 mg orally at 1900h), to 10 normal men. These data indicate that α 1 adrenoceptors are involved in the physiological control of melatonin secretion in man.

Inhibition of reproduction in rams by long daylengths and the acute effect of superior cervical ganglionectomy

Six castrated Soay rams were given testosterone implants, and exposed to an artificial lighting regimen of alternating 16-week periods of long days (16L:8D) and short days (8L:16D) for over 1 year. Under these conditions, the blood plasma concentration of testosterone was maintained relatively constant (4.3--6.4 ng/ml), while the levels of LH and prolactin varied in relation to the alterations in the photoperiod; low levels of LH occurred during long days when the rams were hyperprolactinaemic. During the last period of exposure to long days, 4 of the rams were cranially sympathectomized by removing the superior cervical ganglia. The effect of long daylengths on the secretion of LH and prolactin was blocked; prolactin concentrations began to decline after 2 weeks and LH levels increased. Ganglionectomy also influenced the plasma levels of melatonin; after the operation there was no longer a consistent 24-h rhythm in melatonin values and the increase associated with darkness was abolished. This results supports the view that the regulation of melatonin secretion is involved in the photoperiodic control of reproduction in rams.