Photoperiodic Mediation of Seasonal Breeding and Immune Function In Rodents: A Multi-Factorial Approach

SYNOPSIS. Winter is energetically-demanding; thermoregulatorydemands increase when food availability usually decreases. Physiologicaland behavioral adaptations, including termination of breeding,have evolved among nontropical animals to cope with winter energyshortages. Presumably, selection for mechanisms that permitphysiological and behavioral anticipation of seasonal ambientchanges have led to current seasonal breeding patterns for manypopulations. Energetically—challenging winter conditionscan directly induce death via hypothermia, starvation, or shock;surviving these demanding conditions likely evokes significantstress responses. The stress of coping with energetically-demandingconditions may increase adrenocortical steroid levels to theextent that immune function is compromised. Individuals wouldenjoy a survival advantage if seasonally-recurring stressorscould be anticipated and countered by shunting energy reservesto bolster immune function. The primary environmental cue thatpermits physiological anticipation of season is daily photoperiod,a cue that is mediated by melatonin. However, other environmentalfactors, such as low food availability and ambient temperatures,may interact with photoperiod to affect immune function anddisease processes. Laboratory studies of seasonal changes inmammalian immune function consistently report that immune functionis enhanced in short day lengths. Prolonged melatonin treatmentmimics short days, and also enhances immune function in rodents.In sum, melatonin may be part of an integrative system to coordinatereproductive, immunologic, and other physiological processesto cope successfully with energetic stressors during winter.Social factors influence immune function and changes in socialinteractions may also contribute to seasonal changes in immunefunction. The mechanisms by which social factors are transducedinto immune responses are largely unspecified. In order to understandthe optimization of immune function it is necessary to understandthe interaction of factors, on both mechanistic and functionallevels, that affect immunity.     

Melatonin enhancement of splenocyte proliferation is attenuated by luzindole, a melatonin receptor antagonist

In addition to marked seasonal changes in reproductive, metabolic, and other physiological functions, many vertebrate species undergo seasonal changes in immune function. Despite growing evidence that photoperiod mediates seasonal changes in immune function, little is known regarding the neuroendocrine mechanisms underlying these changes. Increased immunity in short days is hypothesized to be due to the increase in the duration of nightly melatonin secretion, and recent studies indicate that melatonin acts directly on immune cells to enhance immune parameters. The present study examined the contribution of melatonin receptors in mediating the enhancement of splenocyte proliferation in response to the T cell mitogen Concanavalin A in mice. The administration of luzindole, a high-affinity melatonin receptor antagonist, either in vitro or in vivo significantly attenuated the ability of in vitro melatonin to enhance splenic lymphocyte proliferation during the day or night. In the absence of melatonin or luzindole, splenocyte proliferation was intrinsically higher during the night than during the day. In the absence of melatonin administration, luzindole reduced the ability of spleen cells to proliferate during the night, when endogenous melatonin concentrations are naturally high. This effect was not observed during the day, when melatonin concentrations are low. Taken together, these results suggest that melatonin enhancement of splenocyte proliferation is mediated directly by melatonin receptors on splenocytes and that there is diurnal variation in splenocyte proliferation in mice that is also mediated by splenic melatonin receptors.     

Immunoregulatory role of melatonin in cancer

Melatonin is a ubiquitous indole amine that plays a fundamental role in the regulation of the biological rhythm. Disrupted circadian rhythm alters the expression of clock genes and deregulates oncogenes, which finally promote tumor development and progression. An evidence supporting this notion is the higher risk of developing malignancies among night shift workers. Circadian secretion of the pineal hormone also synchronizes the immune system via a reciprocal association that exists between the immune system and melatonin. Immune cells are capable of melatonin biosynthesis in addition to the expression of its receptors. Melatonin induces big changes in different immune cell proportions, enhances their viability and improves immune cell metabolism in the tumor microenvironment. These effects might be directly mediated by melatonin receptors or indirectly through alterations in hormonal and cytokine release. Moreover, melatonin induces apoptosis in tumor cells via the intrinsic and extrinsic pathways of apoptosis, while it protectsthe immune cells. In general, melatonin has a profound impact on immune cell trafficking, cytokine production and apoptosis induction in malignant cells. On such a basis, using melatonin and resynchronization of sleep cycle may have potential implications in immune function enhancement against malignancies, which will be the focus of the present paper.     

Melatonin,immunity and cost of reproductive state in male European starlings.

The effects of reproductive condition and exogenous melatonin on immune function were investigated in castrated European starlings, Sturnus vulgaris. Photorefractory and photostimulated starlings exposed to long days were implanted with melatonin or with blank capsules. Photostimulated starlings with blank capsules exhibited reduced splenocyte proliferation in response to the T-cell mitogen, concanavalin A, compared with the other long-day birds. Exogenous melatonin prevented the suppression of immune function by photostimulation. Photorefractory starlings, with or without melatonin implants, exhibited enhanced immune function compared with photostimulated starlings implanted with blanks. This enhancement was not mediated by endogenous melatonin, but appeared to be related to changes in reproductive state. In addition to the traditional costs of reproduction in birds (e.g. raising of young), there may be a cost of the reproductive state of starlings (i.e. whether they are photorefractory or photostimulated). These data are, we believe, the first to indicate a direct effect of reproductive state on immune function that is independent of both photoperiod (i.e., changes in the duration of melatonin secretion) and gonadal steroids.     

The pars tuberalis as a target of the central clock

The pars tuberalis (PT) of the pituitary has emerged from being a gland of obscure and unknown function to a tissue of central importance to our understanding of how photoperiod regulates seasonal responses. The discovery of melatonin receptors on this gland first pointed to its involvement in seasonal physiology. However, the more recent demonstration of the expression of clock genes in the PT, such as Per1, has heightened interest in the gland. Recent work shows how photoperiod, through the hormone melatonin, affects the timing and amplitude of expression of the Per1 gene, as well as other genes such as Icer. The effect of photoperiod and melatonin on the expression of Per1 in the PT is distinct to its effects on the SCN, and this probably reflects distinct functions of the clock genes in the two tissues - acting as part of the biological clock in the SCN, but as an interval timing system within the PT. The changes in amplitude of Per1 gene expression in response to altered length of photoperiod have provided the first clues as to how the durational melatonin signal is decoded within the neuroendocrine system.     

Seasonal and diurnal melatonin production in exercising sled dogs

Melatonin is a hormone that is released from the pineal gland into the blood stream and is controlled by nerve impulses from the suprachiasmatic nuclei. Melatonin synthesis, which is inhibited by light on the mammalian retina, peaks in plasma concentrations during the night. Though still a subject of intense research, melatonin in mammals is known to effect the reproductive system, thyroid function, and adaptations to seasonal changes. Sled dogs in Fairbanks, Alaska (65 degrees N) can be exposed to anywhere from 21 h of daylight in the summer to 4 h in the winter. While light may be the primary factor influencing melatonin production, we hypothesized that exercise may also affect melatonin production. In the current study, sled dogs were used to study seasonal and diurnal variation in melatonin production. Sled dogs by nature are elite athletes and therefore exercise was a focus in the study. Both exercise and non exercise dogs from 2 distinct latitudes were used. The peak in melatonin production was prolonged in high latitude dogs (65 degrees N), compared with lower latitude dogs (45 degrees N). Dogs at both latitudes show a reduction in peak melatonin levels with exercise, and winter melatonin levels in both locations were higher than the summer. Surprisingly, sled dogs in Alaska had lower melatonin levels than sled dogs in New York.     

Circadian disorganization in experimental arthritis

This review discusses the experimental evidence indicating that arthritis disrupts circadian organization, which was mainly derived from animal studies employing Freund's complete mycobacterial adjuvant (FCA). The defense response to antigenic challenge, mediated in part by cytokines, includes changes in chronobiological central nervous system function, like depressed daily activity, superficial sleep or anorexia. Interferon (IFN)-gamma receptors are detectable in the central circadian pacemaker, the hypothalamic suprachiasmatic nuclei, at a time when the capacity for photic entrainment of the pacemaker became established. The disruptive effects of the systemic injection of IFN on the circadian rhythms of locomotor activity, body temperature and clock-gene mRNA expression have been documented. In the last few years we have examined a number of immune and neuroendocrine circadian rhythms in FCA-injected rats, both in the preclinical phase of arthritis (2-3 days after FCA injection) as well as in the acute phase of the disease (18 days after FCA injection). In arthritic rats, the 24-hour organization of immune and neuroendocrine responses becomes altered. A hormonal pathway involving the circadian secretion of melatonin and a purely neural pathway including, as a motor leg, the autonomic nervous system innervating the lymph nodes were identified. The significant effects of the immune-mediated inflammatory response on the diurnal rhythmicity of adenohypophysial and hypophysiotropic hormones occurred in arthritic rats. Melatonin treatment prevented the alteration in 24-hour rhythms of serum ACTH, prolactin and luteinizing hormone in rats injected with FCA. In addition, melatonin pretreatment prevented the alteration in the 24-hour variation in hypothalamic serotonin and dopamine turnover during the preclinical phase of Freund's adjuvant arthritis in rats. Some pinealectomy-induced immune changes in arthritic rats were also prevented by physiological concentrations of melatonin. Melatonin may play the role of an 'internal synchronizer' for the immune system.     

Melatonin: Nature's most versatile biological signal?

Melatonin is a ubiquitous molecule and widely distributed in nature, with functional activity occurring in unicellular organisms, plants, fungi and animals. In most vertebrates, including humans, melatonin is synthesized primarily in the pineal gland and is regulated by the environmental light/dark cycle via the suprachiasmatic nucleus. Pinealocytes function as 'neuroendocrine transducers' to secrete melatonin during the dark phase of the light/dark cycle and, consequently, melatonin is often called the 'hormone of darkness'. Melatonin is principally secreted at night and is centrally involved in sleep regulation, as well as in a number of other cyclical bodily activities. Melatonin is exclusively involved in signaling the 'time of day' and 'time of year' (hence considered to help both clock and calendar functions) to all tissues and is thus considered to be the body's chronological pacemaker or 'Zeitgeber'. Synthesis of melatonin also occurs in other areas of the body, including the retina, the gastrointestinal tract, skin, bone marrow and in lymphocytes, from which it may influence other physiological functions through paracrine signaling. Melatonin has also been extracted from the seeds and leaves of a number of plants and its concentration in some of this material is several orders of magnitude higher than its night-time plasma value in humans. Melatonin participates in diverse physiological functions. In addition to its timekeeping functions, melatonin is an effective antioxidant which scavenges free radicals and up-regulates several antioxidant enzymes. It also has a strong antiapoptotic signaling function, an effect which it exerts even during ischemia. Melatonin's cytoprotective properties have practical implications in the treatment of neurodegenerative diseases. Melatonin also has immune-enhancing and oncostatic properties. Its 'chronobiotic' properties have been shown to have value in treating various circadian rhythm sleep disorders, such as jet lag or shift-work sleep disorder. Melatonin acting as an 'internal sleep facilitator' promotes sleep, and melatonin's sleep-facilitating properties have been found to be useful for treating insomnia symptoms in elderly and depressive patients. A recently introduced melatonin analog, agomelatine, is also efficient for the treatment of major depressive disorder and bipolar affective disorder. Melatonin's role as a 'photoperiodic molecule' in seasonal reproduction has been established in photoperiodic species, although its regulatory influence in humans remains under investigation. Taken together, this evidence implicates melatonin in a broad range of effects with a significant regulatory influence over many of the body's physiological functions.     

Dexamethasone modulates melatonin MT2 receptor expression in splenic tissue and humoral immune response in mice

Melatonin modulates immune function through its membrane-bound MT1 and MT2 receptors in mammalian system. Adrenal glucocorticoid, an important metabolic hormone is known as a immuno-compromising agent. In the present study, we investigated the effect of dexamethasone on melatonin receptor proteins in spleen tissue and anti-klh-IgG response in Swiss albino mice. Melatonin treatment increased the MT1 and MT2 receptor proteins and anti-klh-IgG than control mice. Dexamethasone treatment increased MT2 receptor protein and anti-klh-IgG than melatonin-treated group. Dexamethasone treatment to melatonin-treated mice showed additive effects and maximally increased the anti-klh-IgG than other experimental groups. A decrease in glucocorticoid receptor (GR) protein was noted in melatonin treated as well as dexamethasone-treated mice. Dexamethasone significantly increased MT2 melatonin receptor protein in spleen and anti-klh-IgG and additively increased anti-klh-IgG when supplemented along with melatonin. Therefore, the present study may suggest that dexamethasone increased humoral immune response permissively by enhancing MT2 receptor expression in splenic tissue of mice.     

Peripheral melatonin modulates seasonal immunity and reproduction of Indian tropical male bird Perdicula asiatica

Seasonal changes in pineal function are well coordinated with seasonal reproductive activity of tropical birds. Further, immunomodulatory property of melatonin is well documented in seasonally breeding animals. Present study elucidates the interaction of peripheral melatonin with seasonal pattern of immunity and reproduction in Indian tropical male bird Perdicula asiatica. Significant seasonal changes were noted in pineal, testicular and immune function(s) of this avian species. Maximum pineal activity along with high immune status was noted during winter month while maximum testicular activity with low immune status was noted in summer. During summer month's long photoperiod suppressed pineal activity and high circulating testosterone suppressed immune parameters, while in winter short photoperiod elevated pineal activity and high circulating melatonin maintained high immune status and suppressed gonadal activity. Therefore, seasonal levels of melatonin act like a major temporal synchronizer to maintain not only the seasonal reproduction but also immune adaptability of this avian species.     

Minireview: Versatile Melatonin: A Pervasive Molecule Serves Various Functions in Signaling and Protection

Melatonin, which is able to enter all tissues and all compartments of the cell, acts in a highly pleiotropic fashion. Some melatonin effects are mediated by membrane receptors, others are receptor independent. Melatonin is produced in the pineal gland and various extrapineal organs of vertebrates, but is also found in invertebrates, angiosperms, and unicells. In mammals, melatonin elicits various secondary humoral responses, e.g., in the immune system via interleukin-4 and other cytokines and in the brain by modulation of NO formation. Melatonin is also a powerful radical scavenger, terminating free radical reaction chains initiated by photooxidants, hydroxyl or peroxyl radicals. The protective potency of this indoleamine is demonstrated by various experiments.     

Minireview: Versatile Melatonin: A Pervasive Molecule Serves Various Functions in Signaling and Protection

Melatonin, which is able to enter all tissues and all compartments of the cell, acts in a highly pleiotropic fashion. Some melatonin effects are mediated by membrane receptors, others are receptor independent. Melatonin is produced in the pineal gland and various extrapineal organs of vertebrates, but is also found in invertebrates, angiosperms, and unicells. In mammals, melatonin elicits various secondary humoral responses, e.g., in the immune system via interleukin-4 and other cytokines and in the brain by modulation of NO formation. Melatonin is also a powerful radical scavenger, terminating free radical reaction chains initiated by photooxidants, hydroxyl or peroxyl radicals. The protective potency of this indoleamine is demonstrated by various experiments.     

褪黑素和褪黑素受体对下丘脑-垂体-肾上腺轴作用的研究进展

松果体于儿童中期可发育至最高峰,普遍在7岁之后开始呈逐渐萎缩,并在成年后逐渐有钙盐沉着。褪黑素主要是由松果体进行合成和分泌所形成,存在较好的昼夜节律性,且通常是通过下丘脑的视交叉上核进行控制,并与环境中的光-暗呈现的周期改变存在密切关联。此外,褪黑素具有极其广泛的生物学作用,且其发挥作用的首站便是与特异性褪黑素受体相关结合,随后经由信号转导系统发挥相应的生物效应。褪黑素受体属于G蛋白耦联受体超家族重要成员之一,其主要是通过百日咳毒素敏感G蛋白的一致性G蛋白通路,减少环腺苷酸的急剧或(和)抑制毛喉菇素刺激的环腺苷酸升高,从而间接影响黑色素活动。下丘脑-垂体-肾上腺轴(HPA轴)是机体在发生应激反应过程中具有一定影响的系统,其所分泌的激素也会表现出昼夜节律性的改变,且此种改变与褪黑素的有关变化呈现出明显的相反性。提示了两者可能存在一定的相关,在机体免疫功能的调控中扮演着不同的角色。本文通过阐述褪黑素和褪黑素受体对HPA轴作用的最新研究进展,旨在明确三者存在的错综复杂的相互作用关系,继而为机体免疫功能调控的一系列疾病研究提供参考依据。     

The changing biological roles of melatonin during evolution: from an antioxidant to signals of darkness,sexual selection and fitness

Melatonin is a molecule present in a multitude of taxa and may be ubiquitous in organisms. It has been found in bacteria, unicellular eukaryotes, macroalgae, fungi, plants and animals. A primary biological function of melatonin in primitive unicellular organisms is in antioxidant defence to protect against toxic free radical damage. During evolution, melatonin has been adopted by multicellular organisms to perform many other biological functions. These functions likely include the chemical expression of darkness in vertebrates, environmental tolerance in fungi and plants, sexual signaling in birds and fish, seasonal reproductive regulation in photoperiodic mammals, and immunomodulation and anti‐inflammatory activity in all vertebrates tested. Moreover, its waning production during aging may indicate senescence in terms of a bio‐clock in many organisms. Conversely, high melatonin levels can serve as a signal of vitality and health. The multiple biological functions of melatonin can partially be attributed to its unconventional metabolism which is comprised of multi‐enzymatic, pseudo‐enzymatic and non‐enzymatic pathways. As a result, several bioactive metabolites of melatonin are formed during its metabolism and some of the presumed biological functions of melatonin reported to date may, in fact, be mediated by these metabolites. The changing biological roles of melatonin seem to have evolved from its primary function as an antioxidant.     

New insights into ancient seasonal life timers

Organisms must adapt to seasonal changes in the environment and time their physiology accordingly. In vertebrates, the annual change in photoperiod is often critical for entraining the neuroendocrine pathways, which drive seasonal metabolic and reproductive cycles. These cycles depend on thyroid hormone (TH), reflecting its ancestral role in metabolic control. Recent studies reveal that - in mammals and birds - TH effects are mediated by the hypothalamus. Photoperiodic manipulations alter hypothalamic TH availability by regulating the expression of TH deiodinases (DIO). In non-mammalian vertebrates, light acts through extraretinal, 'deep brain' photoreceptors, and the eyes are not involved in seasonal photoperiodic responses. In mammals, extraretinal photoreceptors have been lost, and the nocturnal melatonin signal generated from the pineal gland has been co-opted to provide the photoperiodic message. Pineal function is phased to the light-dark cycle by retinal input, and photoperiodic changes in melatonin secretion control neuroendocrine pathway function. New evidence indicates that these comparatively divergent photosensensory mechanisms re-converge in the pars tuberalis of the pituitary, lying beneath the hypothalamus. In all vertebrates studied, the pars tuberalis secretes thyrotrophin in a light- or melatonin-sensitive manner, to act on neighbouring hypothalamic DIO expressing cells. Hence, an ancient and fundamentally conserved brain thyroid signalling system governs seasonal biology in vertebrates.     

Photoperiod and melatonin influence seasonal gonadal cycles in the grasshopper mouse (Onychomys leucogaster)

Development of the reproductive apparatus was delayed in grasshopper mice maintained from birth in short photoperiods (10 h light/day). The inhibitory effects of short photoperiods on sexual maturation eventually waned and mice in 10L:14D became reproductively active. Adult mice transferred from long (14 h light/day) to short photoperiods underwent testicular regression after 10 weeks and complete gonadal redevelopment after 30 weeks. A similar phenomenon was observed in adult female mice; oestrous cycles ceased within 3 weeks and resumed after 13 weeks in the short photoperiod. The regressive effects of short photoperiods on the male reproductive system were mimicked by daily injections of melatonin administered to mice housed in 14L:10D. Responsiveness of the female reproductive system to melatonin was reduced among photorefractory as compared to photosensitive mice. We suggest that the initial rate of sexual maturation and the timing of seasonal breeding in adult mice are regulated by photoperiod; effects of short daylengths on the neuroendocrine-reproductive axis appear to be mediated by the pineal gland.     

褪黑激素受体基因的研究进展

褪黑激素通过与药理学特异性的高亲和性G-蛋白耦联受体相结合来发挥其生物学功能。本文介绍了褪黑激素受体的结构、功能与调控、褪黑激素受体基因的克隆及基因结构、褪黑激素受体基因的发育性表达与作用、褪黑激素受体基因的定位与多态性分析,并讨论了该基因与繁殖季节性的关系。 Abstract:Melatonin exerts its biological effects through pharmacological specific,high affinity G protein-coupled receptors.This review introduced the structure,function,and regulation of melatonin receptor,the cloning and structure,developmental expression,mapping and polymorphism of melatonin receptor gene.The relationship between melatonin receptor gene and reproductive seasonality was also discussed.     

Seasonal variations of gonadotropins and prolactin in the laboratory rat. Role of maternal pineal gland

The laboratory rat, a non-photoperiodic rodent, exhibits seasonal fluctuations of melatonin. Melatonin has been found to be readily transferred from the maternal to the fetal circulation. No data exist on the possible influence of maternal pineal gland upon seasonal variations of the offspring. The aim of the present study was to asses the influence of the maternal melatonin rhythm on the offspring postnatal development of the reproductive hormones LH, FSH and prolactin. Male offspring from control, pinealectomized (PIN-X) and PIN-X + melatonin (PIN-X+MEL) mother Wistar rats were studied at 21, 31, and 60 days of age. Seasonal age-dependent variations were found for all hormones studied in control offspring but PIN-X offspring showed a tendency to have reduced duration or altered seasonal variations. Maternal melatonin treatment to PIN-X mothers partially restored the effect of pinealectomy. The chronological study of LH, FSH, and prolactin in PIN-X offspring also showed an altered pattern as compared to control-offspring. Melatonin treatment to the mothers partially restored the developmental pattern of reproductive hormones. Results of this study indicate that maternal pineal gland of the laboratory rat is involved in the seasonal postnatal development variations of reproductive hormones of the offspring.     

Pineal modulation of thymus and immune function in a seasonally breeding tropical rodent, Funambulus pennanti

The immune system driven by cytokines is now known to be influenced by various other endocrine glands and its hormones. Results of the present study indicate a bidirectional relation between the pineal-thymus axis and the immune system status of an Indian tropical rodent, Funambulus pennanti, during winter months (reproductive inactive phase), when it faces maximum challenges from nature. Pinealectomy during the reproductive inactive phase inhibited thymus and spleen functions, which resulted in significant changes in leukocyte and lymphocyte counts and T-cell-mediated immune function (measured in terms of delayed-type hypersensitivity response to oxazolone). Blastogenic responses of lymphoid cells (thymocytes, splenocytes, and lymph node cells) also decreased following ablation of the pineal gland. To check the definite role of the pineal gland we injected melatonin into pinealectomized squirrels, and the suppressed immune function was significantly restored. Neuroendocrine control of the pineal gland on the histocompatible tissues in this seasonal breeder, F. pennanti, suggests an adaptive mechanism of the immune system for survival in the tropical zone. J. Exp. Zool. 289:90-98, 2001.     

The pineal organ of teleost fishes

The pineal organ of teleost fish is a directly photosensory organ that contains photoreceptor cells similar to those of the retina. It conveys photoperiod information to the brain via neural pathways and by release of indoleamines, primarily melatonin, into the circulation. The photoreceptor cells respond to changes in ambient illumination with a gradual modulation of neurotransmission to second-order neurons that innervate various brain centres, and by modulation of indoleamine synthesis. Melatonin is produced rhythmically, and melatonin synthesis may be regulated either directly by ambient photoperiod, or by an endogenous circadian oscillator that is entrained by the photoperiod. During natural conditions, melatonin is produced at highest levels during the night. Although the pineal organ undoubtedly influences a variety of physiological parameters, as assessed by experimental removal of the pineal organ and/or administration of exogenous indoleamines, its role in any physiological situation is not clear cut. The effects of any interference with pineal functions appear to vary with the time of year and experimental photothermal regimes. There are strong indications that the pineal organ is one component in a central neural system that constitutes the photoperiod-responding system of the animal, i.e. the system that is responsible for correct timing of daily and seasonal physiological rhythms. It is important to envisage the pineal organ as a part of this system; it interacts with other photosensory structures (the retina, possibly extraretinal non-pineal photoreceptors) and circadian rhythm generators