Melatonin treatment improves the antioxidant status and decreases lipid content in brain and liver of rats.

This study examines the effect of a daily administration of melatonin for 45 days at two doses (0.5 and 1.0 mg/kg body wt.) on antioxidant status, lipid peroxidation and lipid profile in the brain and liver in rats. Both doses of melatonin caused a significant decrease in lipid peroxidation and the levels of cholesterol, phospholipids, triglycerides and free fatty acids in the examined tissues. Concomitantly, the treatment with melatonin augmented the activity of the brain and liver antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase as well as increased glutathione levels. These results offer a support to the hypothesis that melatonin in pharmacological amounts effectively reduces oxidative stress and acts as an antihyperlipidemic agent.

Melatonin decreases the oxidative stress produced by 2,4-dichlorophenoxyacetic acid in rat cerebellar granule cells.

2,4-Dichlorophenoxyacetic acid (2,4-D) is one of the most widely used herbicides due to its relatively moderate toxicity and to its biodegradability in the soil. In toxic concentrations, 2,4-D displays strong neurotoxicity, partly due to generation of free radicals. Since melatonin has remarkable antioxidant properties, the objective of this study was to assess to what extent it was effective in preventing the 2,4-D effect on redox balance of rat cerebellar granule cells (CGC) in vitro. Cellular viability, generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS), reduced glutathione (GSH) levels, and the activities of the antioxidant enzymes Cu/Zn-superoxide dismutase (Cu/Zn-SOD), Mn-SOD, selenium-glutathione peroxidase (Se-GPx) and catalase (CAT) were measured in CGC exposed to 2,4-D and/or melatonin for 48 h. In CGC cultures exposed to 2,4-D, cell viability, GSH levels and CAT activity decreased significantly whereas ROS generation and Se-GPx activities were augmented. Except for Se-GPx activity, all these changes were counteracted by the concomitant addition of 0.1 or 0.5 mM melatonin. In addition, incubation of CGC with melatonin alone resulted in augmentation of cell viability, GSH levels and Se-GPx activity. RNS generation and SOD activity remained unaffected by either treatment. Since melatonin was able to counteract most of redox changes produced by 2,4-D in CGC in culture, the experimental evidence reported further support the efficacy of melatonin to act as a neuroprotector.

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.

Melatonin in Alzheimer's disease and other neurodegenerative disorders.

Increased oxidative stress and mitochondrial dysfunction have been identified as common pathophysiological phenomena associated with neurodegenerative disorders such as Alzheimer's disease (AD), Parkinson's disease (PD) and Huntington's disease (HD). As the age-related decline in the production of melatonin may contribute to increased levels of oxidative stress in the elderly, the role of this neuroprotective agent is attracting increasing attention. Melatonin has multiple actions as a regulator of antioxidant and prooxidant enzymes, radical scavenger and antagonist of mitochondrial radical formation. The ability of melatonin and its kynuramine metabolites to interact directly with the electron transport chain by increasing the electron flow and reducing electron leakage are unique features by which melatonin is able to increase the survival of neurons under enhanced oxidative stress. Moreover, antifibrillogenic actions have been demonstrated in vitro, also in the presence of profibrillogenic apoE4 or apoE3, and in vivo, in a transgenic mouse model. Amyloid-beta toxicity is antagonized by melatonin and one of its kynuramine metabolites. Cytoskeletal disorganization and protein hyperphosphorylation, as induced in several cell-line models, have been attenuated by melatonin, effects comprising stress kinase downregulation and extending to neurotrophin expression. Various experimental models of AD, PD and HD indicate the usefulness of melatonin in antagonizing disease progression and/or mitigating some of the symptoms. Melatonin secretion has been found to be altered in AD and PD. Attempts to compensate for age- and disease-dependent melatonin deficiency have shown that administration of this compound can improve sleep efficiency in AD and PD and, to some extent, cognitive function in AD patients. Exogenous melatonin has also been reported to alleviate behavioral symptoms such as sundowning. Taken together, these findings suggest that melatonin, its analogues and kynuric metabolites may have potential value in prevention and treatment of AD and other neurodegenerative disorders.

Twenty-four hour rhythm of plasma prolactin in female rabbit pups. Correlation with hypothalamic and adenohypophysial dopamine, serotonin, gamma-aminobutyric acid and taurine content.

Lactation in the rabbit is a nocturnal activity, extremely short and regular, that can be a strong synchronizer for the development of circadian rhythmicity in the pups. In the present study, 24-h rhythmicity of plasma prolactin and median eminence and anterior pituitary content of dopamine (DA), serotonin (5HT), gamma-aminobutyric acid (GABA) and taurine were examined in 11 days old female pups kept under 16 h light:8 h dark photoperiods (lights on at 08:00 h). Groups of six to seven female rabbit pups were killed by decapitation at six different time points throughout a 24-h cycle, starting at 09:00 h. Plasma prolactin levels changed significantly throughout the day, showing two peaks, one at first half of rest span (at 13:00 h) and another one at the beginning of the scotophase (at 01:00 h), just preceding doe visit. Median eminence DA content changed in a bimodal way as a function of time of day, displaying two maxima, at the beginning of the rest span and of the activity phase. Median eminence DA and plasma prolactin correlated significantly in an inverse way. Two maxima in median eminence 5HT levels were found, about 4 h in advance to the prolactin peaks. Circulating prolactin correlated inversely with median eminence 5HT content and directly with adenohypophysial 5HT content. Median eminence GABA content reached its maximum at the beginning of the scotophase and correlated significantly with plasma prolactin concentration. A positive correlation between plasma prolactin and adenohypophysial taurine content was observed. These results show that the circadian rhythmicity in prolactin secretory mechanisms in female rabbit pups develops during the early neonatal life.

Comparative effects of melatonin and vitamin E in restoring aortic relaxation in pancreatectomized rats.

In a previous study we reported the efficacy of melatonin to restore the decreased relaxation response to acetylcholine (ACh) or to sodium nitroprusside (SNP) in aortic rings of rats turned hyperglycemic by subtotal pancreatectomy. The effect was amplified by pre-incubation in a high (44 mmol/l) glucose solution, a situation that resulted in oxidative stress. We hereby compare the effect of another antioxidant, vitamin E, with that of melatonin on ACh response in intact aortic rings or on SNP response in endothelium-denuded aortic rings obtained from pancreatectomized or sham-operated rats. Dose-response curves to ACh or SNP were performed in the presence or absence of melatonin or vitamin E (10-5 mol/1) in 10 or 44 mmol/1 glucose medium. Melatonin was more effective than vitamin E in restoring ACh- or SNP-induced relaxation of aortic rings in a high glucose medium. The differences between the two antioxidants may rely on the ability of melatonin to diffuse readily into intracellular compartments.

Melatonin and sleep in aging population.

The neurohormone melatonin is released from the pineal gland in close association with the light-dark cycle. There is a temporal relationship between the nocturnal rise in melatonin secretion and the 'opening of the sleep gate' at night. This association, as well as the sleep promoting effect of exogenous melatonin, implicates the pineal product in the physiological regulation of sleep. Aging is associated with a significant reduction in sleep continuity and quality. A decreased production of melatonin with age is documented in a majority of studies. Diminished nocturnal melatonin secretion with severe disturbances in sleep/wake rhythm has been consistently reported in Alzheimer's disease (AD). A recent survey on the effects of melatonin in sleep disturbances, including all age groups, failed to document significant and clinically meaningful effects of exogenous melatonin on sleep quality, efficiency and latency. However, in clinical trials involving elderly insomniacs and AD patients suffering from sleep disturbances exogenous melatonin has repeatedly been found to be effective in improving sleep. The results indicate that exogenous melatonin is more effective to promote sleep in the presence of a diminished production of endogenous melatonin. A MT1/MT2 receptor analog of melatonin (ramelteon) has recently been introduced as a new type of hypnotics with no evidence of abuse or dependence.

Role of melatonin in neurodegenerative diseases.

The pineal product melatonin has remarkable antioxidant properties. It scavenges hydroxyl, carbonate and various organic radicals, peroxynitrite and other reactive nitrogen species. Melatonyl radicals formed by scavenging combine with and, thereby, detoxify superoxide anions in processes terminating the radical reaction chains. Melatonin also enhances the antioxidant potential of the cell by stimulating the synthesis of antioxidant enzymes like superoxide dismutase, glutathione peroxidase and glutathione reductase, and by augmenting glutathione levels. The decline in melatonin production in aged individuals has been suggested as one of the primary contributing factors for the development of age-associated neurodegenerative diseases, e.g., Alzheimer's disease. Melatonin has been shown to be effective in arresting neurodegenerative phenomena seen in experimental models of Alzheimer's disease, Parkinsonism and ischemic stroke. Melatonin preserves mitochondrial homeostasis, reduces free radical generation, e.g., by enhancing mitochondrial glutathione levels, and safeguards proton potential and ATP synthesis by stimulating complex I and IV activities. Therapeutic trials with melatonin have been effective in slowing the progression of Alzheimer's disease but not of Parkinson's disease. Melatonin's efficacy in combating free radical damage in the brain suggests that it may be a valuable therapeutic agent in the treatment of cerebral edema after traumatic brain injury.

Melatonin, immune function and aging.

Aging is associated with a decline in immune function (immunosenescence), a situation known to correlate with increased incidence of cancer, infectious and degenerative diseases. Innate, cellular and humoral immunity all exhibit increased deterioration with age. A decrease in functional competence of individual natural killer (NK) cells is found with advancing age. Macrophages and granulocytes show functional decline in aging as evidenced by their diminished phagocytic activity and impairment of superoxide generation. There is also marked shift in cytokine profile as age advances, e.g., CD3+ and CD4+ cells decline in number whereas CD8+ cells increase in elderly individuals. A decline in organ specific antibodies occurs causing reduced humoral responsiveness. Circulating melatonin decreases with age and in recent years much interest has been focused on its immunomodulatory effect. Melatonin stimulates the production of progenitor cells for granulocytes-macrophages. It also stimulates the production of NK cells and CD4+ cells and inhibits CD8+ cells. The production and release of various cytokines from NK cells and T-helper lymphocytes also are enhanced by melatonin. Melatonin presumably regulates immune function by acting on the immune-opioid network, by affecting G protein-cAMP signal pathway and by regulating intracellular glutathione levels. Melatonin has the potential therapeutic value to enhance immune function in aged individuals and in patients in an immunocompromised state.

Melatonin effects on behavior: possible mediation by the central GABAergic system.

The best described function of the pineal hormone melatonin is to regulate seasonal reproduction, with its daily production and secretion varying throughout the seasons or the photoperiod. Additionally, a number of behavioral effects of the hormone have been found. This review describes the effects of melatonin in rodent behavior. We focus on: (a) inhibitory effects (sedation, hypnotic activity, pain perception threshold elevation, anti-convulsive activity, anti-anxiety effects); and (b) direct effects on circadian rhythmicity (entrainment, resynchronization, alleviation of jet-lag symptoms, phase-shifting). Most of these effects are clearly time-dependent, with a peak of melatonin activity during the night. One of the possible mechanisms of action for melatonin in the brain is the interaction with the GABAergic system, as suggested by neurochemical and behavioral data. Finally, some pineal hormone effects might be candidates as putative therapies for several human disorders.

Changes in (3H)leucine incorporation into pineal proteins following estradiol or testosterone administration: involvement of the sympathetic superior cervical ganglion.

Pineal denervation by superior cervical ganglionectomy (Gx) decreased high affinity binding of estradiol (E2) to the pineal cytosol of female rats and of testosterone to the cytosol of male rats by 40 and 26% and by 75 and 80%, 5 and 14 days after sugery; hormone binding remained unchanged up to 24 h after surgery. Binding to the nuclear fraction decreased sigificantly by 2 weeks after incorporation of (3H) leucine into pineal proteins in Gx. A single injection of E2 (mug) to testosterone propionate (TP) (500 mug) failed to increase the Gx rats when injected 1 or 5 days after surgery. Significant increases were observed in sham-operated controls or in rats subjected to bilateral decentralization of ganglia; however on the 5th day an impairment was observed in hormone ability to enhance [3H]leucine incorporation in decentralized rats. The administration of isoproterenol 19 and 3 h before sacrifice replenished pineal-binding sites for E2 and testosterone in Gx rats, but failed to restore the responsiveness of denervated pineals to hormone administration. Moreover, E2 or TP treatment blocked the increase in labeled amino acid incorporation into proteins brought about by isoproterenol per se. The administration of propranolol 2 and 7 h after hormone injection decreased the ability of E2 and TP to enhance [3H]leucine incorporation by 55 and 41%, respectively. Tyrosine hydroxylase activity of the superior cervical ganglia decreased by 36 and 41% 6 h after E2 or TP administration, and by 43 and 47% after 3 daily injections of the hormones, whereas pineal tyrosine hydroxylase remained unchanged. Hormone treatment for 3 days increased the in vitro uptake of norepinephrine by the ganglia but did not affect uptake in the pineal gland. These data indicate that the integrity of neurons of the superior cervical ganglia is an absolute requirement for E2 and testosterone to enhance [3H]leucine incorporation into pineal proteins in rats.

[3H]Dihydroergocryptine binding in anterior pituitary and prolactin secretion: further evidence of brain regulation of adenohypophyseal receptors.

Secretion of PRL by the pituitary gland is under tonic inhibitory control by tuberoinfundibular dopaminergic neurons. The aim of the present work was to assess the influence of the hypothalamus on the binding of [3H]dihydroergocryptine, a dopamine agonist, in the adenohypophysis and its role in the hypoprolactinemic action of exogenous dopamine. Serum PRL levels were elevated in ovariectomized adult rats with electrolytic lesions of the median eminence compared to levels in sham-operated animals; [3H]dihydroergocryptine binding to anterior pituitary was elevated from a control value of 161.2 +/- 7.7 to 229.0 +/- 10.0 fmol/mg protein in rats studied 7-21 days after placement of the lesion (P < 0.05). Scatchard analysis of [3H]dihydroergocryptine binding in lesioned and sham-operated animals indicated an increase of 62% in the number of binding sites without a change in binding affinity. In a second group of rats, with or without median eminence destruction, the effects of different doses of ip dopamine on serum PRL were determined. The hypoprolactinemic effect of each dose of the amine was more evident in median eminence-lesioned rats than in sham-operated controls. These results suggest that when hypothalamic influences on the anterior pituitary are removed, there is increased binding of dopamine by the gland and increased hypoprolactinemic effects of the exogenous amine. This indicates that receptors in the pituitary are under hypothalamic control, and this could be an important regulatory mechanism in neuroendocrine events.

Role of prostaglandins in rat pineal neuroeffector junction. Changes in melatonin and norepinephrine release in vitro.

The effects of prostaglandins (PGs) on melatonin secretion and norepinephrine (NE) and release in rat pineal gland were examined in vitro. To study melatonin secretion, pineal explants were incubated for 6 h in tissue culture 199 medium with 1-1000 nM PGE1, PGE2, or PGF2 alpha. melatonin concentration in pineal glands and media was determined by RIA, PGE2 increased pineal and medium melatonin at all concentrations tested, with a maximum of 1 nM; PGE1 was effective only at concentrations 100-1000 times greater, whereas 100 nM PGF2 alpha gland. Exposure of pineal explants to 10 microM NE brought about a 20-fold increase in melatonin release to the medium. This effect was impaired significantly, but not blocked, by prior exposure to indomethacin, acetylsalicylic acid, or mefenamic acid at supramaximal concentrations to inhibit PG synthesis (100 microM). To examine the effects of PGs on NE release, endogenous NE stores in pineal nerve endings were labeled in vitro by incubating rat pineals with [3H]NE for 30 min. Fifty minutes later, at the time when spontaneous radioactivity efflux had leveled off, transmitter release was elicited by a 1-min exposure to 80 mM K+ (S1), and the stimulus was repeated 35 min later (S2). PGs (10-100 nM) were added to the medium 20 min before S2. Ratios between fractional release of the two consecutive stimulations (S2/S1) varied between 0.84 and 1.16 in control pineals. Only 100 nM PGE2 impaired significantly transmitter release by 40%. These results suggest that PGE2 can play a role in NE-stimulated melatonin synthesis. At greater concentrations PGE2 inhibits NE release from pineal nerve endings.

Effect of inferior laryngeal nerve section on thyroid function in rats.

To examine the role of thyroid parasympathetic innervation in organ's function, rats subjected to inferior laryngeal nerve (ILN) section were employed. This procedure decreased thyroid [3H]choline uptake by about half. Bilaterally ILN-sectioned rats treated with methylmercaptoimidazole for 4 days exhibited a significant impairment of the methylmercaptoimidazole-induced goitrogenic response. Unilateral ILN section resulted in further atrophy of the ipsilateral thyroid lobe in hypophysectomized rats. One week after ILN section a significant decrease of serum T4 and an increase of serum TSH were observed. Bilateral ILN section generally decreased circulating T4 for up to 28 days after surgery, while a unilateral ILN section caused a transient T4 decrease for 1 week after surgery. Compensatory thyroid growth in rats subjected to unilateral thyroidectomy (hemi Tx) and ILN section performed ipsilaterally to the remaining lobe, was significantly smaller than that of rats subjected to hemi Tx alone. Hemi Tx depressed serum T4 and increased serum TSH levels significantly. These hormonal changes were prevented by unilateral superior cervical ganglionectomy (SCGx), but were unaffected by ILN section. The combination of SCGx and ILN section negated the facilitating effect of SCGx on thyroid secretion and impaired the increase in compensatory thyroid growth brought about by thyroid sympathetic denervation. Thyroid mitotic index studies in hemi Tx rats receiving ILN section, SCGx, or a combination of both indicated that the increase in the number of follicular mitosis caused by hemi Tx was significantly impaired by ILN section and was significantly increased by SCGx. SCGx potentiation of thyroid follicular mitotic activity was partially prevented by concomitant ILN section. These results support a significant role of thyroid sympathetic and parasympathetic innervation in the control of organ's growth and secretory activity.

Inhibition of human platelet aggregation and thromboxane-B2 production by melatonin: evidence for a diurnal variation.

The effects of melatonin on platelet aggregation and thromboxane-B2 (TxB2) production induced by 1-4 x 10(-6) M adenosine diphosphate (ADP) or 0.6 x 10(-3) M arachidonic acid (AA) were assessed in platelet-rich plasma (PRP). Micromolar concentrations of melatonin inhibited in a dose-dependent way ADP-induced platelet aggregation with individual inhibitions 40% or more at 10(-6)-10(-5) M. A significant depression of AA-induced platelet aggregation was observed only at 10(-5)-10(-4) M melatonin. Morning (0830 h)-evening (1800 h) studies of ADP-induced platelet aggregation in seven normal men showed a higher sensitivity at 1800 h when analyzed as a global inhibitory effect of melatonin (P less than 0.01). Moreover, only during the evening hours did melatonin induce reversible aggregation, an index of inhibition of the platelet secretory process elicited by ADP exposure. No diurnal variability in melatonin inhibition of AA-induced aggregation was detected. TxB2 production elicited by AA in the evening was inhibited significantly in a concentration-related manner by a 2-min preincubation with 10(-9)-10(-5) M melatonin, while during the morning hours the inhibition was significant only at 10(-6) M or higher melatonin concentrations. In the case of ADP, the inhibition of TxB2 release attained significance at 10(-5)-M (0830 h) or 10(-6)-M concentrations (1800 h). In the presence of either stimulatory agent, melatonin depression of TxB2 generation was about 2-fold greater at 1800 h than at 0830 h. The diurnal changes in melatonin effect on TxB2 production were also observed in thrombin-stimulated washed platelets. The present data indicate the existence of circadian variations in platelet responsiveness to melatonin in humans.

Effect of pentoxifylline on alpha- and beta-adrenoceptor sites in cerebral cortex medial basal hypothalamus and pineal gland of the rat.

The intraperitoneal injection of the methylxanthine derivative pentoxifylline (3,7-dimethyl-1-(5-oxo-hexyl)-xanthine] brought about, 3 hr later, a significant depression of alpha- and beta-adrenoceptor sites in the cerebral cortex, and of beta-adrenoceptor sites in medial basal hypothalamus and pineal gland, (assessed from the specific binding of radioactive dihydroergocryptine and dihydroalprenolol respectively). The changes in the density of binding sites were not accompanied by significant modifications of the Kd's. Sympathetic denervation of the pineal gland by superior cervical ganglionectomy (SCGx) abolished the changes of beta-adrenoceptor number in the pineal caused by pentoxifylline. The increase of alpha-adrenoceptor sites in the hypothalamus brought about by ganglionectomy was not affected by injection of pentoxifylline. Pentoxifylline did not compete in vitro for radioligand binding to brain membranes. These results suggest that methylxanthines depress brain adrenoceptor sites acutely, probably by down-regulation of receptors following the increase in catecholamine release caused by injection of the drug.

Differential effect of interferon-gamma injection on mitogenic responses in sympathetically denervated submaxillary lymph nodes of rats.

This study aimed to assess the effect of interferon (IFN)-gamma treatment on mitogenic responses in submaxillary lymph nodes in the presence or absence of local sympathetic nerves. Adult male rats were subjected to a unilateral superior cervical ganglionectomy and to a contralateral sham-operation. Seven days later, rats received five i.p. daily injections of human IFN-gamma (10(5) U.I./kg) or saline. On the day after the last injection, rats were killed at six different times throughout a 24-h cycle and the mitogenic effect of lipopolysaccharide (LPS) and concanavalin A (Con A) was assessed in single-cell suspensions of lymph nodes. In vehicle-treated rats, proliferation responses to LPS in innervated lymph nodes did not show time-of-day variations while those in denervated lymph nodes attained a maximum at 17:00 h. Following IFN-gamma administration, a promoting effect of LPS mitotic response was detected at 01:00 h at the innervated side only. As far as the mitogenic responses to Con A, proliferation in innervated lymph nodes of vehicle-treated controls attained a maximum at 09:00 h. Such a daily variation in response to Con A was not detectable at the denervated side. IFN-gamma treatment increased significantly Con A activity by promoting a greater mitogenic response at 01:00 h. Sympathetic denervation of lymph nodes brought about a shift in the maximum in number of cells per mg of lymph node from 21:00 to 13:00 h. After IFN-gamma treatment, maxima in cell number occurred at 05:00 h at both the innervated and denervated side. The results indicate that IFN-gamma effects in rat submaxillary lymph nodes are under substantial modulation by local sympathetic nerves.

Failure of melatonin to increase serum prolactin levels in ovariectomized rats subjected to superior cervical ganglionectomy or pinelaectomy.

The effects of melatonin on serum prolactin levels were examined in ovariectomized rats primed with oestradiol and progesterone, and subjected to bilateral superior cervical ganglionectomy or pinealectomy. Ganglionectomy resulted in a significant depression of the serum prolactin concentrations, as well as in impairment of the prolactin release evoked by administration of steroid. Treatment with melatonin increased serum prolactin in control but not in ganglionectomized rats. Injection of melatonin protentiated the steroid-induced release of prolactin in control rats; this effect of melatonin was not detected in ganglionectomized rats. Pinealectomy did not affect basal prolactin levels, nor impair the release of prolactin evoked by steroid treatment; however, it was effective in blocking the melatonin-induced release of prolactin in vehicle-treated rats, as well as the potentiation of steroid-induced prolactin release by melatonin. Intracranial surgery by itself increased prolactin release. These results suggest that systemically administered melatonin needs an intact pineal gland to augment serum prolactin levels.

Basic aspects of melatonin action.

Melatonin is synthesized and secreted during the dark period of the light-dark cycle. Thus, melatonin has an obvious association with sleep, at least in diurnal animals. Rhythmic nocturnal melatonin secretion is directly generated by the circadian clock, located in mammals within the suprachiasmatic nuclei (SCN), and is entrained to a 24-h period by the light-dark cycle. The periodic secretion of melatonin may be used as a circadian mediator to any system than can "read" the message. In addition, direct effects of the hormone on the SCN could explain some of melatonin effects on the circadian system. Duration of melatonin nocturnal secretion is directly proportional to the length of the night and it has been demonstrated experimentally to be the critical parameter for photoperiod integration. The two main hypotheses to explain the action of melatonin are the duration hypothesis (supporting that night length is coded by the duration of the melatonin secretory phase) and the coincidence hypothesis (holding that physiological responses are linked to the existence of a diurnal rhythm in sensitivity to melatonin). The sites and mechanisms of action of melatonin for circadian and photoperiodic responses are far from being elucidated, but action through specific membrane receptor sites is well documented. In view of melatonin s lipophilic nature, interactions with specific intracellular proteins like calmodulin or tubulin, or with nuclear receptor sites, have also been considered, whereas the physiological significance of the documented antioxidant effect of melatonin remains to be settled. Melatonin seems to act as an "arm" of the circadian clock, giving a time-related signal to a number of body functions; one of these, the circadian organization of an organism's defence, is discussed in some detail as an example.

Changes of prolactin regulatory mechanisms in aging: 24-h rhythms of serum prolactin and median eminence and adenohypophysial concentration of dopamine, serotonin, (gamma-aminobutyric acid, taurine and somatostatin in young and aged rats.

Twenty-four hour rhythmicity of serum prolactin and median eminence and anterior pituitary content of dopamine (DA), serotonin (5HT), gamma-aminobutyric acid (GABA), taurine and somatostatin were examined in 2 months-old and 18-20 months-old Wistar male rats. The concentration of prolactin was higher in aged rats, with peaks in both groups of rats at the early phase of the activity span. Median eminence DA content of young rats attained its maximum at the middle of rest span and decreased as prolactin levels augmented while the lowest values of adenohypophysial DA were observed at the time of prolactin peak. DA rhythmicity disappeared in aged rats. GABA content of median eminence and adenohypophysis was lower in aged rats, with maximal values of median eminence GABA at light-dark transition in young rats and at the second half of activity span in aged rats. Serum prolactin correlated positively with median eminence GABA in young rats and negatively with pituitary GABA in young and aged rats. Median eminence somatostatin peaked at the beginning of the activity phase (young rats) or at the end of the rest phase (aged rats). Prolactin levels and somatostatin content correlated significantly in young rats only. Median eminence and pituitary 5HT and taurine content did not change with age. The results indicate disruption of prolactin regulatory mechanisms with aging in rats.