Lower Melatonin Concentrations Research Articles

Several antioxidant pathways are involved in astrocyte protection by melatonin.

Neuroprotection provided by melatonin has been shown to be more relevant in vivo than in neuronal cultures. Given the role of astrocytes in neuronal support and protection, studies were initiated to elucidate the possible protective effect of the antioxidant melatonin against oxidative stress in these cells. Both low and high concentrations of melatonin were able to protect astrocytes with even higher efficiency than the known antioxidant glutathione (GSH). The mechanisms involved may be different for high (1 mm) and low (100 nm) concentrations of the indole. The GSH cycling appeared not to be involved in the protection at high doses. High doses of melatonin neither influenced GSH levels nor gene expression for the several antioxidant enzymes studied; thus, melatonin's protective effect was likely because of its free radical scavenging action in this case. However, melatonin concentrations in the nanomolar range require the presence of GSH to be effective. No increase in GSH synthesis was found, but low doses of melatonin increased gene expression and activity of glutathione peroxidase. As this enzyme requires GSH as substrate to be active, this may be the reason why the effect of this melatonin concentration is GSH dependent. In vivo, melatonin levels exhibit a wide range of concentrations with much lower levels in the blood and significantly higher concentrations in other body fluids and within cells. Thus, melatonin may normally function as an indirect and direct antioxidant in vivo.

Physiological concentrations of melatonin inhibit the nitridergic pathway in the Syrian hamster retina.

In the present work, the effect of melatonin on the hamster retinal nitridergic pathway was examined. When the retinas were incubated in the presence of low concentrations (1 pM-10 nM) of melatonin for 15 min, a significant decrease of nitric oxide synthase (NOS) activity was observed. However, when crude retinal homogenates were preincubated with melatonin for 15 min, no changes in NOS activity were detected, despite the fact that under the same conditions trifluoperazine, a calmodulin inhibitor, significantly decreased enzymatic activity. Kinetic analysis showed that melatonin decreased the V(max) of retinal NOS without changes in the K(m). On the other hand, low concentrations (100 pM) of melatonin significantly reduced retinal L-arginine influx. A decrease in the V(max) of L-arginine uptake was observed in the presence of melatonin, whereas the K(m) remained unchanged. Melatonin significantly inhibited the accumulation of cyclic guanosine monophosphate (cGMP) levels induced by both L-arginine and sodium nitroprusside (SNP). In summary, the present results indicate that melatonin could be a potent inhibitor of the retinal nitridergic pathway.

Hypothalamic involvement in chronic migraine

OBJECTIVESChronic migraine (CM), previously called transformed migraine, is a frequent headache disorder that affects 2%-3% of the general population. Analgesic overuse, insomnia, depression, and anxiety are disorders that are often...

An endogenous 5‐HT7 receptor mediates pigment granule dispersion in Xenopus laevis melanophores

Melatonin (5-methoxy N-acetyltryptamine) and serotonin (5-HT) exert rapid, but opposite effects on pigment granule distribution in Xenopus laevis melanophores. Low concentrations of melatonin (10(-11) - 10(-9) M) cause a dramatic perinuclear aggregation of the melanin-containing granules, while 5-HT (10(-8) - 10(-5) M) disperses pigment granules throughout the cell. The present study found that pharmacological doses of melatonin (> or =10(-6) M) induced a time- and concentration-dependent pigment granule dispersion, which was mediated by an endogenous melanophore 5-HT receptor. 5-HT produced a concentration-dependent elevation of melanophore cyclic AMP, and 5-HT-induced dispersion was blocked by H89 (10(-4) M), an inhibitor of protein kinase A (PKA), but not by a PKC inhibitor (Ro 31-8220, 10(-5) M), indicating a vital role for cyclic AMP in 5-HT-induced dispersion. 5-HT-mediated dispersion was not blocked by antagonists selective for G(s)-coupled 5-HT(4) (GR113808) or 5-HT(6) (Ro 04-6790, Ro 63-0563, olanzepine) receptors, nor by 5-HT(1 - 3) (pindolol, ketanserine, metoclopramide, MDL72222, tropisetron) receptor antagonists, but was inhibited by a selective 5-HT(7) receptor antagonist, DR4004, and other antagonists with a high affinity for 5-HT(7) receptors. The rank order of antagonist potency was: risperidone (mean pK(B) 7.82)>methiothepin (7.43)>DR4004 (6.92)>mesulergine (6.83)>methysergide (6.60)>[+/-]-sulpiride (5.81)>spiperone (5.52). The agonist potency order [mean pEC(50), 5-CT (8.68)>5-HT (7.13)>5-MT (6.94)>8-OH-DPAT (4.79)>sumatriptan (<4)] was also consistent with an action on 5-HT(7) receptors. RT - PCR confirmed that melanophores express 5-HT(7) receptor mRNA. The pigment dispersing effect of high melatonin concentrations in melanophores is most likely mediated by activation of 5-HT(7) receptors. Conceivably some of the effects attributed to pharmacological doses of melatonin in mammals may be mediated by activation of 5-HT(7) receptors.

The influence of different concentrations of melatonin on the cell surface hydrophobic characteristics of Neisseria meningitidis.

The cell surface hydrophobicity of micro-organisms is a characteristic that has been associated with the colonization of mammalian epithelia and with their capacity to induce diseases. Melatonin is a hormone produced by the pineal gland that affects the immune response mechanism. This study investigated, as an expression of the virulence of Neisseria meningitidis, how its hydrophobic characteristics were affected by exposure to increasing concentrations of melatonin. An increase in the cell surface hydrophobicity of N. meningitidis was found at concentrations of 1 mmol l(-1), while lower concentrations of melatonin did not significantly affect this particular cell surface characteristic of the micro-organism. It may be concluded that melatonin clearly influences the cell surface hydrophobicity of N. meningitidis, a circumstance that should be taken into account in future studies to determine whether this hormone plays a role in the variable pathogenicity of the bacteria in different hosts.

The effect of melatonin on cellular activation processes in human blood.

The pineal hormone melatonin, due to its lipophilic nature, has access to every cell and every part of a cell in the body, suggesting that it could exert effects on blood immune cells. The regulation of the activation of monocytes may be important in a number of diseases, especially pathophysiological conditions associated with inflammatory reactions. Considering this, a study on the effect of melatonin on monocytes in whole blood was carried out. Melatonin added at a final concentration of 5 ng/mL to whole blood in vitro reduced lipopolysaccharide (LPS)-induced tissue factor (TF) activity in monocytes by 55% in blood from a group of subjects with melatonin-sensitive cells. At even lower concentrations of melatonin (20-50 pg/mL) and in the physiological range, a trend of suppressed LPS-induced TF activity by approximately 20% was seen. A further indication of a downregulation of LPS-stimulated monocytes by melatonin was shown by its reduction of LPS-induced tumor necrosis factor (TNF). Twenty to one hundred pg/mL melatonin caused a significant reduction of LPS-induced TNF production by approximately 25-30%. In contrast, melatonin at a final concentration of 10 pg/mL, added to whole blood incubated with LPS and also the phorbol ester, PMA, caused a significant rise of 25%; whereas 100 pg/mL enhanced LPS + PMA-induced TNF by approximately 80% as compared to LPS + PMA alone. These effects were not detectable during the winter darkness of Tromsø (70 degrees N), probably due to the high content of melatonin in the blood even at daytime. These results show that melatonin may have a beneficial effect by suppressing the expression of TF activity in LPS-stimulated monocytes. Furthermore, the results indicate that LPS-induced TF in monocytes of whole blood is independent of protein kinase C (PKC) activation. Melatonin is probably amplifying cellular activation reactions that are PKC-dependent. This may be physiologically important in upregulation of the immune system.

Effect of continuous melatonin infusions on steady-state plasma melatonin levels in rats under near physiological conditions.

It was the aim of this study to measure the actual amount of melatonin required for elevating the circulating hormone from low daytime levels to the 10-fold higher nocturnal steady-state concentrations in rats. For this purpose, escalating doses of melatonin were continuously infused into the right jugular vein and blood samples were repeatedly drawn from the left jugular vein for a period of 2 hr in freely moving catheterized rats. In order to achieve an about 10-fold elevation of the plasma melatonin concentration, 500 ng melatonin/hr had to be infused, i.e., about 300 times the normal nocturnal melatonin content of the pineal. Infusions of up to 61 ng melatonin/hr (equivalent to the melatonin content of 40 pineals at darkness) failed to cause a significant rise of the low daytime steady-state concentrations in the blood. If the dose of 500 ng melatonin/h was infused at night, a less-pronounced rise of the blood levels was observed, as compared to that caused by the infusion of the same dose during daytime. No differences were found in the rate of metabolism between daytime and nighttime. The results of this study indicate 1) that the low basal concentrations of melatonin in the blood are not affected by an increased melatonin supply up to a certain critical threshold, 2) that the rat pineal gland would have to release all its melatonin content almost every 10 sec in order to sustain the elevated steady-state level of melatonin in the circulation during the dark period, and 3) that significant day/night differences exist in the disposition of circulating melatonin if administered in near physiological amounts and under near physiological conditions.

Melatonin suppression of PC12 cell growth and death

Melatonin has previously been reported to influence cell differentiation and growth in a number of cell culture systems in vitro. In this paper, we describe the effects of high pharmacological and low physiological concentrations of melatonin on cell growth in rat pheochromocytoma cells (PC12 cells). Melatonin produced a biphasic response with respect to cell growth in PC12 cells. At low concentrations (1–10 nM) melatonin suppressed PC12 cell growth whereas at higher concentration (10 μM) it prevented cell death. Cultures treated with high concentrations of melatonin displayed an increase in cell number and a decreased release of lactic acid dehydrogenase (LDH) into the culture media, indicating that melatonin was enhancing cell survival as opposed to stimulating cell proliferation. Inhibition of cell death by high concentrations of melatonin was both time and concentration-dependent and did not require the continued presence of melatonin throughout the entire time of incubation. These studies suggest melatonin is preventing either apoptosis or programmed cell death. In contrast, concentrations of melatonin (1–10 nM) at or near the binding affinity for the nuclear receptor, RZR β, suppressed PC12 cell growth. At these concentrations, melatonin failed to inhibit forskolin-induced cAMP formation and process outgrowth as well as prevent forskolin suppression of cell growth. These data indicate that PC12 cells probably lack functionally active cell surface receptors for melatonin and suggest the interaction of melatonin with the nuclear receptor may be responsible for suppression of PC12 cell growth.

Case-control study of evening melatonin concentration in primary insomnia.

The function of melatonin is not fully established, but recent studies suggest that it plays a role in the regulation of sleep. Thus, physiological doses of melatonin given to healthy volunteers decreased the time taken to fall asleep,1 and the incidence of insomnia in the population rises during middle and old age,2 when serum concentrations of melatonin decline.3 Haimov et al found that elderly patients with insomnia had lower than normal peak urinary concentrations of the melatonin metabolite 6-sulphatoxy melatonin and a delayed onset to peak secretion.4 We investigated evening plasma melatonin concentrations in subjects with primary insomnia and matched controls and predicted that the subjects with insomnia would have lower melatonin concentrations. Cases and controls were recruited predominantly by advertisement, but two cases …

Determination of melatonin in blood plasma, using capillary gas chromatography and electron impact medium-resolution mass spectrometry.

Determination of the low concentrations (less than 10-100 pg ml-1) of melatonin in blood plasma by gas chromatography/mass spectrometry has previously required the use of negative ionization (electron capture) mass spectrometry, a technique particularly delicate to set up. We show that a general-purpose medium-resolution instrument gives adequate sensitivity in electron impact mode, when used with a capillary gas chromatographic column-switching device. A straightforward extraction procedure gives good specificity, and the limit of useful measurement is better than 10 pg per 1 ml sample.

Melatonin in serum and urine in patients with idiopathic pain syndromes

In 16 healthy volunteers, 16 patients with neurogenic pain syndromes, 37 patients with idiopathic pain syndromes, and 30 depressed patients, melatonin in serum was determined at 2 a.m. when the peak concentration was expected. In a somewhat larger series comprising 53 healthy volunteers, 14 patients with neurogenic pain syndromes, and 35 patients with idiopathic pain syndromes, melatonin was measured in urine collected during the night in a standardized manner. Chronic pain patients (with neurogenic or idiopathic pain disorders) and depressed patients had significantly lower melatonin in serum at 2 a.m. than healthy volunteers. Chronic pain patients also had significantly lower melatonin in urine than healthy volunteers, even when age, sex, and body weight were taken into account. The low melatonin concentrations were related to increased depressive symptomatology, especially sadness, bodily discomfort, inner tension, concentration difficulties, and pain. As low concentrations of melatonin in serum and urine also are found in patients with depressive disorders, the results are in line with the suggestion that the chronic idiopathic pain syndrome may be a variant of depressive disease, or the two syndromes may share a common pathogenic mechanism.

Beta-adrenergic binding is increased by melatonin and alpha-adrenergic compounds

Binding of the beta-adrenergic ligands [3H]dihydroalprenolol and [125I]cyanopindolol to pineal particulate fractions was increased 1- to 3.5-fold by addition of low concentrations of melatonin, alpha-adrenergic agonists, or alpha-adrenergic antagonists. Minimum concentrations of melatonin or alpha-adrenergic compounds which increased beta-adrenergic binding were between 1 pM and 0.1 nM. The increased binding of [3H]dihydroalprenolol caused by melatonin (0.1 muM) was attributed to a major increase in Bmax, which persisted in protein fractions after removal of melatonin. Melatonin enhancement of [3H]dihydroalprenolol binding was apparent after 5 to 7 min (30(0], was was optimal between 20 and 40 min, and decreased at longer times. Alpha-Adrenergic receptors are unchanged during beta-receptor enhancement.

Validation of a sensitive direct assay for melatonin for investigation of circadian rhythms in different species.

The role of melatonin in animals which do not show marked seasonal changes in reproduction is disputed, in part because of the wide variation in reported concentrations. One reason for this may be the difficulties associated with the measurements of low molar concentrations of melatonin and the presence of a wide variety of potentially cross-reacting substances. The availability of a high affinity antiserum has allowed an assay, with low cross-reactivity and good sensitivity, to be established for the direct measurement of melatonin in a wide range of biological fluids, in particular serum, plasma and follicular fluid from man and rat. The high affinity of the antiserum enabled a tritium label of high specific activity to be used, removing the problems associated with the iodination of a small molecular weight compound. Melatonin concentrations in the assay were evaluated by four different methods: UV absorbance, gas chromatography, comparison of the immunoreactive concentrations of the label with the expected concentration by dilution and by comparison with a previously established assay which uses the same antiserum. Melatonin was measured in serum from twelve healthy women over two 24-h periods; eight women with normal menstrual cycles and four taking the contraceptive pill. Concentrations were found to range from 19.8 to 215 pmol/l during the day in both groups. In women with normal menstrual cycles peak concentrations of 513.2 +/- 54.1 (S.E.M.) pmol/l were recorded at 04.00 h, whereas higher concentrations were found in women taking the pill, reaching a peak of 849.12 +/- 21.8 (S.E.M.) pmol/l at 04.00 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of daily melatonin injections on female reproduction in the white-footed mouse, Peromyscus leucopus

Seventy laboratory-reared (16L:8D at 21°) Peromyscus leucopus were injected daily for 7 weeks with either saline, 0.5, 5.0, or 50 μg of melatonin at either 2 or 12 hr into the photophase. Mice injected with 50 μg at 12 hr exhibited a substantial decrease ( P < 0.01) in relative reproductive tract weight (2.3 ± 0.4 mg/g body wt) when compared to saline injected animals (4.6 ± 0.9). Seven out of the nine melatonin-treated mice also exhibited an imperforate vagina. A lesser effect was noted at lower melatonin concentrations. No regression occurred in mice injected 2 hr into the photophase. Thermoregulatory characters (nesting, brown fat, winter molt) were not affected by melatonin injections. In a second experiment 49 mice were injected with 50 μg of melatonin at either 7.5, 10, 14, 15, 18, or 23 hr after lights on. A gradual antigonadal effect was observed with a peak during late photophase. Mice injected during early scotophase (18 hr) exhibited little antigonadal effects of melatonin, but animals injected at 23 hr had reproductive tract weights which were considerably less ( P < 0.05).

Melatonin in amniotic fluid during human parturition.

Concentrations of melatonin have been measured in amniotic fluid obtained during late pregnancy and labour. Samples obtained by amniotomy during labour contained higher concentrations of melatonin than samples taken at amniotomy for the induction of labour. Amniotic fluid obtained by amniocentesis in late pregnancy contained lower concentrations of melatonin than amniotic fluid obtained by amniotomy. The implications of these findings are discussed in relation to pineal influences on parturition.

The effect of light and dark background upon melatonin levels in larval Xenopus

1. 1. Body melanophore numbers and spectrophotofluorometric determinations of melatonin and 5-hydroxytryptamine were made on whole Xenopus laevis larvae reared in aquaria with light and dark surfaces. 2. 2. Light background reared larvae had lower melatonin concentrations and fewer melanophores. 3. 3. Dark background reared larvae had higher melatonin concentrations and more melanophores. 4. 4. 5-Hydroxytryptamine concentrations were not significantly different in the two groups.