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Biomedical subjects

J M Míguez

Publications and source records attributed to J M Míguez.

17 recordsLinked to original sources

Response of plasma and gastrointestinal melatonin, plasma cortisol and activity rhythms of European sea bass (Dicentrarchus labrax) to dietary supplementation with tryptophan and melatonin.

Melatonin is an effective antioxidant, immunostimulant, gonadal maturating regulator and antistress indoleamine that may be potentially useful for fish farmers. We have explored two possible ways of increasing plasma melatonin levels through the diet: direct melatonin supplementation (ME diet) and supplementation with the melatonin precursor tryptophan (TRP diet). To this end, a group of sea bass was fed a commercial diet (STD diet) at a regular time for 16 days, after which plasma, intestine, and bile samples were taken at four different time points: 120 min before, and 15, 180 and 480 min after feeding. Locomotor activity, intestinal and biliary melatonin, and plasma melatonin, serotonin and cortisol levels were measured. This same sampling process and analyses were also carried out after feeding sea bass TRP diet or ME diet for 1 week. Our results show that melatonin, but not tryptophan supplementation of the diet increases plasma, intestine and bile levels of melatonin. Plasma serotonin levels, on the other hand, were increased by dietary tryptophan, but not by melatonin, confirming the availability of supplemented tryptophan for serotonin synthesis. Both treatments were equally effective in reducing the high cortisol levels observed with the STD diet.

Animals↗

Glucagon effects on brain carbohydrate and ketone body metabolism of rainbow trout.

The levels of glycogen in brain, lactate and acetoacetate in brain and plasma, glucose in plasma and the activities of brain key enzymes of glycogen metabolism (glycogen phosphorylase, GPase, glycogen synthetase, GSase), gluconeogenesis (fructose 1,6-bisphosphatase, FBPase), and glycolysis (6-phosphofructo 1-kinase, PFK) were evaluated in rainbow trout, Oncorhynchus mykiss, from 0.5 to 3 hr after intraperitoneal injection of 1 ml/kg(-1) body weight of saline alone (controls) or containing bovine glucagon at three different doses: 10, 50, and 100 ng/g(-1) body weight. The results obtained demonstrate, for the first time in a teleost fish, the existence of changes in brain carbohydrate and ketone body metabolism following peripheral glucagon treatment. A clear stimulation of brain glycogenolytic potential was observed after glucagon treatment, as judged by the time- and dose-dependent changes observed in brain glycogen levels (up to 88% decrease), and GPase (up to 30% increase) and GSase (up to 42% decrease) activities. In addition, clear time- and dose-dependent increased and decreased levels were observed in brain of glucagon-treated rainbow trout for lactate (up to 60% increase) and acetoacetate (up to 67% decrease), respectively. In contrast, no significant changes were observed after glucagon treatment in those parameters related to glycolytic/gluconeogenic capacity of rainbow trout brain. Altogether, these in vivo results suggest that glucagon may play a role (direct or indirect) in the regulation of carbohydrate and ketone body metabolism in brain of rainbow trout.

Acetoacetates↗

Selective changes in the contents of noradrenaline, dopamine and serotonin in rat brain areas during aging.

This study examines the age-associated changes in noradrenaline (NA), dopamine (DA), 3,4-dihydroxyphenyl-acetic acid (DOPAC), serotonin (5-HT) and 5-hydroxy-3-indoleacetic acid (5-HIAA) in different brain areas of rats. DA and DOPAC concentrations in striatum increased at third month of age, remaining without significant variations until 12th month of age, and decreasing in 24-month-old rats. DA concentration dropped in hippocampus, amygdala and brainstem of 24-month-old-rats, whereas DOPAC levels decreased only in hippocampus. These changes suggest an age-dependent deficit of the dopaminergic system, presumably related to a reduced number/activity of DA nigrostriatal and mesolimbic neurons. An age-induced decline in NA content was found in the pons-medulla, the area containing NA neuronal bodies. Concentrations of 5-HT were reduced with aging in frontal cortex, showing a tendency to decrease in all brain areas examined. The increased 5-HIAA/5-HT ratio found in frontal cortex, amygdala and striatum suggests an age-related decreased synthesis and an accelerated 5-HT metabolism. The 5-HIAA content decreased in brainstem of the oldest rats. These findings point to a selective impairment of nigrostriatal and mesolimbic DA in aging rats, whereas reductions in NA were restricted to cell bodies region and 5-HT showed changes of different extent in areas of terminals and neuronal cell bodies.

Aging↗

Changes with age in daytime and nighttime contents of melatonin, indoleamines, and catecholamines in the pineal gland: a comparative study in rat and Syrian hamster.

Previous studies in rodents showed a severe deterioration of pineal physiology with aging. The present study investigated the age-related changes in the content of monoamines and metabolites in rat and Syrian hamster pineal gland. In addition to melatonin, the levels of 5-hydroxytryptophan (5HTP), serotonin (5HT), 5-hydroxyindoleacetic acid (5HIAA), N-acetylserotonin (N-Ac-SHT), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), and noradrenaline (NA) were measured by HPLC. Pronounced reductions were found in 5HT and 5HIAA contents during daytime in rats of 24 months, which had not been observed in animals of 12 months. In addition, nighttime pineal 5HIAA, N-Ac-5HT, and melatonin contents were decreased in the old rats, although a significant day:night variation persisted. Also a diurnal fluctuation in NA, DA, and DOPAC contents was present in young and middle-aged rats but not for NA and DOPAC in the oldest rats due to a decrease in the nighttime levels. Pineal DA levels were also reduced in 24-month-old rats during the night, although a marked day:night change was still found. In the Syrian hamster pineal, significant reductions in daytime 5HT and 5HIAA were found respectively at 12 and 18 months, while nighttime levels of these compounds were decreased from 18 months. The nocturnal content of N-Ac-5HT dropped gradually from 12 months, and melatonin was reduced by 74% and 86% in hamsters of 18 and 24 months, respectively. In all these compounds, a significant day:night variation was observed irrespective of age. However, neither a day:night variation nor an effect of aging was found in terms of pineal NA content. In contrast, pineal DA and DOPAC levels displayed a diurnal variation in hamsters of 1.5 and 6 months, but not in animals of 12 and 18 months due a reduced nighttime content. These data suggest that the decline of pineal melatonin with age is a consequence of a deficit in the pathway of serotonin utilization. This probably is explained by a reduced N-acetyltransferase activity, which may be linked to impaired pineal catecholaminergic neurotransmission.

3,4-Dihydroxyphenylacetic Acid↗

Synchronizing circadian rhythms in early infancy.

Entrainment to the 24-hour light-dark cycle is of adaptive significance to mammals. Human infants are no exception, but some postnatal care habits prevalent in developed countries can interfere with the physiological mechanisms underlying circadian synchronization. We describe the physiological mechanisms of entrainment to the light-dark cycle in fetuses and newborns, and some common parental care behaviors which subject the developing circadian system of the newborn to conflicting temporal cues. Improvements in parental care are proposed which may improve the circadian synchronization of newborns, and their parents or caregivers.

Animals↗

Melatonin effects on serotonin synthesis and metabolism in the striatum, nucleus accumbens, and dorsal and median raphe nuclei of rats.

This work examined the influence of the pineal gland and its hormone melatonin on the metabolism of serotonin (5-HT) in discrete areas of the forebrain, such as the striatum and the nucleus accumbens, and the midbrain raphe. The content of 5-HT and its major oxidative metabolite, the 5-hydroxyindoleacetic acid (5-HIAA), as well as the in-vivo tryptophan hydroxylation rate were examined after long-term pinealectomy (one month) and daily melatonin treatment (500 micrograms/kg; twice daily for ten days) in pinealectomized rats. Pinealectomy did not alter 5-HT content in any of these brain areas, but it significantly increased the content of 5-HIAA in striatum and the 5-HIAA/5-HT ratio in nucleus accumbens. The normal values of these parameters were recuperated after administration of exogenous melatonin, but it also increased the rate of tryptophan hydroxylation in both areas. In addition, melatonin treatment decreased the levels of 5-HIAA in dorsal raphe nucleus. These data suggest that the pineal gland, through the secretion of melatonin, modulates the local metabolism of 5-HT in forebrain areas by acting on the oxidative deamination. Moreover, melatonin injected in pinealectomized rats derives in a more extended effect than pinealectomy and induces a stimulation of 5-HT synthesis in the striatum, probably due to a pharmacological effect. These results point to the striatum as a target area for the interaction between pineal melatonin and the serotonergic function, and suggest a differential effect of the melatonin injected on areas containing serotonergic terminals and cell bodies, which may relevant for the mode of action of melatonin and its behavioral effects.

Animals↗

Melatonin receptors in the suprachiasmatic nuclei and pars tuberalis of testosterone induced photoresponsive rats.

The sexual axis of rats can be rendered photoresponsive by testosterone implants. We have studied in these conditions whether rat pars tuberalis (PT) melatonin receptor density would be decreased after exposure to short photoperiod (SP) like as observed in long day seasonal breeders. The answer is no, but we observed that testosterone induced a photoperiod independent decrease in PT melatonin receptor density. These results show that testosterone-induced photosensitivity in rat is not linked to an SP-induced decrease in PT melatonin receptor density. However, testosterone regulates PT melatonin receptors independently of the photoperiod.

Animals↗

Diurnal changes in the content of indoleamines, catecholamines, and methoxyindoles in the pineal gland of the Djungarian hamster (Phodopus sungorus): effect of photoperiod.

Previous studies in Syrian hamster have shown that the correlations between the daily fluctuations in the contents of pineal indoleamines and methoxyindoles are influenced by the photoperiod, and that dopamine may play a role in the regulation of pineal function. The present study investigated the 24 hour changes in the content of 5-hydroxytryptophan (5-HTP), serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), N-acetylserotonin (NAS), melatonin, 5-methoxytryptophol (5-MT), noradrenaline (NA), dopamine (DA), and 3,4-dihydroxyphenylacetic acid (DOPAC) in the pineal gland of female Djungarian hamsters exposed to long (LP; LD 16:8) or short (SP; LD 10:14) photoperiods for 10 weeks. Pronounced nocturnal increases of N-acetylserotonin and melatonin content were observed irrespective of the photoperiod regime. The content of 5-HT was markedly decreased during the first hour of the night in LP, which contrasted with the lack of changes in NAS and melatonin content at this time. In SP, an increased 5-HTP content and a less obvious decrease in 5-HT content was observed during the night, although melatonin and NAS content were even higher than in LP. Similar daily patterns as for 5-HT were observed in the 5-HT oxidative metabolites (5-HIAA and 5-MT). When considering values throughout the day, a poor correlation for 5-HT vs.. NAS and melatonin content was observed, which was particularly evident during the nighttime. These data indicate that the daily variation of pineal indoles may not only be dependent on changes in the N-acetyltransferase activity but also in other mechanisms regulating pinealocyte 5-HT availability. As previously reported in other species of hamster, pineal NA content did not show daily variations in LP conditions, although a nocturnal increase was detected in SP. In contrast, pronounced nocturnal increases were observed in the content of DA and its acid metabolite, DOPAC, irrespective of the photoperiod. These data indicate the existence of an increased dopaminergic turnover during the night and further support a role for DA in the regulation of melatonin synthesis and in the synchronization of the pineal functions.

Animals↗

Daily and photoperiodic melatonin binding changes in the suprachiasmatic nuclei, paraventricular thalamic nuclei, and pars tuberalis of the female Siberian hamster (Phodopus sungorus).

Using quantitative autoradiography, 2-(125)I-melatonin binding was investigated throughout the light:dark cycle in the suprachiasmatic nuclei (SCN), paraventricular nuclei (PVT), and pars tuberalis (PT) of adult female Siberian hamsters kept for 10 weeks in either long or short photoperiods (LP or SP, respectively). Plasma melatonin concentrations were measured by radioimmunoassay, and the sexual status of the animals was established by visual inspection of vaginal smears and by weighing uteri after sacrifice. The SCN displayed neither daily nor photoperiod-dependent variations in specific binding. Melatonin receptors in these nuclei would be regulated neither by plasma melatonin nor by the light:dark cycle or sexual steroids. By contrast, melatonin receptor density in the PT displayed both strong daily (maximal values during the first half of the light period and minimal values during the night) and photoperiod-dependent (maximal values in LP) variations. These variations dependent on changes in the maximal binding (Bmax) without differences in the dissociation constant (Kd). Daily melatonin receptor densities in the PT of LP- and SP-exposed animals might be regulated by the dark:light transition but not by melatonin. Daily profiles of 2-(125)I-melatonin-specific binding in the PT were independent of photoperiod. Factors underlying the photoperiod-related variations presently are unknown. Concerning the PVT, weak variations in specific binding were detected in SP only when time points were grouped according to the light or dark periods. It is not yet possible to conclude whether they have any physiological relevance. These results show clearly that the regulation of melatonin receptors varies among structures (SCN, PVT, and PT) in the Siberian hamster and is also totally different from that found in the rat.

Analysis of Variance↗

Changes in pineal indoleamines in rats after single melatonin injections: evidence for a diurnal sensitivity to melatonin.

We recently determined that melatonin stimulated serotonin (5-HT) secretion from rat pineal glands by increasing 5-HT release from the pinealocytes (microM melatonin concentrations) and by inhibiting 5-HT uptake in the pineal sympathetic nerve endings (mM melatonin concentrations). The present study investigated whether a single melatonin injection could alter the content of indoleamines in the rat pineal gland, as well as its possible dependence on the daytime of administration. Melatonin (150 micrograms/kg) was i.p. injected at 8 time points (11.00 h, 14.00 h, 17.00 h, 20.00 h, 23.00 h, 02.00 h, 05.00 h and 08.00 h) to rats kept in 12:12 h light:dark cycle (lights on at 07.00 h). Melatonin injections in the afternoon (17:00 h) and late in the nighttime (02.00 h and 05.00 h) decreased pineal 5-HT content 90 min later. The levels of 5-hydroxyindoleacetic acid (5-HIAA) were also decreased 90 min after the melatonin treatment at 14.00 h, 17.00 h and 02.00 h. The effect of melatonin on 5-HT content was a long-fasting effect (still evident after 180 min) only when injected at 02.00 h, whereas 5-HIAA levels were found to be decreased 180 min after melatonin treatment at 14.00 h and 23.00 h. No changes in these compounds were detected 240 min after melatonin treatment. Moreover, melatonin did not change 5-hydroxytryptophan levels at any of the daytime points studied. By contrast, 90 min after the injection of melatonin at 20.00 h, an increased content of pineal N-acetylserotonin was observed. This effect of melatonin could be mediated through a phase alteration of the pineal N-acetyltransferase activity rhythm by acting on the suprachiasmatic clock, although a direct melatonin effect on the pineal rhythmic function cannot be excluded. The effects of the hormone on 5-HT and 5-HIAA contents agree with previous findings on the inhibitory effect of pharmacological doses of melatonin on pineal 5-HT uptake, which presumably would result in a decreased intraneuronal content of 5-HT and its acid metabolite. These data point to an acute regulatory action of exogenous melatonin on the pineal melatonin synthesis pathway which seems to be limited to two daytime phases; the afternoon-early evening period and the second half of the night.

5-Hydroxytryptophan↗

Changes in the hypothalamic serotonergic function may mediate the endocrine effects of melatonin.

The effects of a single injection of melatonin on serum thyroxine, corticosterone and prolactin levels, and the associated changes in the metabolism of serotonin (5-HT) in several hypothalamic regions of male rats kept under a 14-h light 10-h dark cycle (lights on at 08:00 h), are investigated. Melatonin (500 micrograms/kg, s.c.) or replacing saline was injected at 18:00 h, and 8 animals from treated and control groups were killed 1 h (19:00 h), 12 h (06:00 h) and 18 h (12:00 h) later. Melatonin significantly reduced serum thyroxine, corticosterone and prolactin levels within 1 h of treatment, indicating the existence of an acute inhibitory effect on hormonal secretion. Serum prolactin levels were increased 18 h after treatment, suggesting the implication of a different long-term regulatory mechanism. Injected melatonin induced several acute changes in the metabolism of 5-HT in the hypothalamus. Thus, a significant increase of 5-HT content in the preoptic area-anterior hypothalamic (POA-AH) and medial hypothalamic regions of melatonin-treated rats was observed within 1 h of treatment. The content of 5-hydroxyindoleacetic acid (5-HIAA) increased in medial hypothalamus, and the ratio 5-HIAA/5-HT decreased in POA-AH. The results show that melatonin acutely modifies the serotonergic activity in POA-AH and medial hypothalamus, and simultaneously inhibits thyroid, adrenal and pituitary hormonal secretion. The results are discussed suggesting that the hypothalamic serotonergic system could be an ideal mechanism underlying short-term melatonin effects on endocrine function.

Animals↗

Effect of streptozotocin-induced diabetes mellitus on serotonin measures of peripheral tissues in rats.

The present study was conducted to examine whether experimental diabetes (streptozotocin-induced) promotes changes in serotonin (5HT) measures of peripheral tissue. Platelet-free plasma 5HT, tryptophan and 5-hydroxyindolacetic acid (5HIAA), whole blood 5HT and renal, liver, intestinal and lung 5HT and 5HIAA levels were measured in rats of four experimental groups: control, diabetic, diabetic+insulin and non-diabetic+insulin. Several serotonin measures were unaltered in all four experimental groups, i.e. plasma, liver and lung 5HT and 5HIAA levels. Whole blood 5HT levels descended about 50% in diabetic rats, then recovered their proper levels after 1 week of insulin therapy. Diabetic animals had a significantly greater intestinal 5HT concentration (+50% versus control), while intestinal 5HIAA levels did not achieve statistical significance despite a -26% decrement in their value. Both renal 5HT and 5HIAA levels were decreased in diabetic animals and recovered with insulin therapy. Peripheral tissue 5HT measures were not varied by insulin administration to non-diabetic animals. The results are consistent with a 5HT release, which is diminished in enterochromaffin cells and enhanced in platelet concomitantly to a minor platelet 5HT uptake, for explaining alterations of plasma/blood 5HT measures in experimental diabetes, and with a diminished synthesis of 5HT for explaining renal changes.

Animals↗

Platelet serotonin transport is altered in streptozotocin-induced diabetic rats.

The present work was conducted to examine whether experimental diabetes (streptozotocin-induced) promotes changes in mean platelet volume, and platelet serotonin (5HT) uptake and content. These variables were measured in from four experimental groups: control, diabetic, diabetic+insulin, and non-diabetic+insulin. Animals treated fifteen days before with streptozotocin had platelets with higher 5HT uptake affinity, 5HT content, and volume. The insulin therapy reestablished the control values of all of these three variables. Non-diabetic animals treated one week with insulin did not show any variations. The effects of in vitro application of insulin, hyperglycaemic incubation medium, and streptozotocin on platelet amine uptake and release were also examined. Only those platelets incubated with streptozotocin showed an altered platelet 5HT uptake. No changes were observed for spontaneous 5HT release. The results are consistent with: a) an increase of platelet uptake capacity, as a consequence of an increase in platelet turnover, for explaining alterations of intraplatelet 5HT contents in experimental diabetes; b) a non-direct effect of insulin and glucose levels on platelet 5HT uptake -for explaining its dysfunctions in experimental diabetes-; c) the contribution of alterations in platelet 5HT transport for explaining the higher incidence of vascular complications in diabetic patients; d) the suitability of platelet as a model for investigating neuronal 5HT reuptake.

Animals↗

Evidence for a regulatory role of melatonin on serotonin release and uptake in the pineal gland.

Melatonin has been proposed to exert some regulatory actions within the pineal gland itself. The present study examined the effect of melatonin on the release of serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) from rat pineal glands by using an in vitro perifusion system. Melatonin induced a concentration-dependent stimulatory effect on 5-HT secretion from 10(-6) M to 10(-3) M. Maximal effects were obtained with melatonin 10(-3) M and concentrations lower than 10(-6) M were without effect. The secretion of 5-HIAA was inhibited by melatonin 10(-3) and 10(-4) M, but it was increased when pineals were incubated with 10(-5) and 10(-6) M of melatonin. The indoleamine secretion was also studied on peripherally denervated rat pineal glands. Basal output of 5-HT from these glands was increased when compared with those from control rats. In contrast, the secretion of 5-HIAA was strongly reduced after removal of the sympathetic input to the pineal gland. Melatonin 10(-3) M failed to stimulate 5-HT release from denervated pineal glands, although it inhibited 5-HIAA secretion. In contrast, melatonin 10(-5) M enhanced 5-HT release without altering 5-HIAA output. Fluoxetine, a 5-HT uptake inhibitor, produced similar effects than mM concentrations of melatonin on the indoleamine secretion from control pineal glands, but it had no effect on glands taken from peripherally denervated rats. These data suggest that mM concentrations of the pineal hormone are able to stimulate 5-HT release from the pinealocyte, while mM concentrations of melatonin increase extracellular 5-HT by inhibiting its reuptake in the adrenergic nerve endings. These findings are discussed in relation to the possible role of melatonin regulating the intra- and extracellular availability of 5-HT in the pineal gland and its significance as an autocrine factor.

Animals↗

Melatonin effect on serotonin uptake and release in rat platelets: diurnal variation in responsiveness.

UNLABELLED: The present study was conducted to examine whether melatonin impairs serotonin (5HT) release and uptake in rat platelets. Exposure of platelet-rich plasma samples (PRP) to melatonin induced a concentration-dependent inhibition of 5HT uptake and the value of IC50 was 1.3 x 10(-3) M. We have also investigated the melatonin effect on the kinetic parameters of platelet 5HT uptake. Transport capacity was inhibited (Vmax; CONTROL: 2.28 +/- 0.52, Melatonin: 0.74 +/- 0.13 pmol/10(7) platelet.min; p < 0.05) while the affinity of 5HT for its uptake carriers remained unaltered, thus indicating a non-competitive effect. Studies carried out to determine the existence of a differential morning (8:00h)-evening (21:00h) melatonin effect showed a higher platelet uptake sensitivity at 8:00h (two-way ANOVA, p < 0.001). Spontaneous 5HT release was not impared by the hormone and no daily variation in sensitivity was detected. The possible mechanism of action of melatonin on platelet transport is discussed, and the results support the suitability of the platelet model for studying sensitivity changes in target cells to the hormone.

Analysis of Variance↗

Analysis of adrenergic regulation of melatonin synthesis in Siberian hamster pineal emphasizes the role of HIOMT.

Seasonal variations of environmental factors are translated into annual fluctuations in synthesis and release of melatonin, which in turn acts as a neuroendocrine messenger for the synchronization of annual functions. So far, most studies performed to understand the regulation of melatonin synthesis have used the non seasonal laboratory rat. It was demonstrated that nocturnal melatonin synthesis depends on alpha- and beta-adrenergic activation of the enzyme arylalkylamine N-acetyltransferase (AA-NAT). In this study, we investigated the mechanisms of melatonin synthesis in the Siberian hamster, a seasonal species with marked photoperiodic variation in melatonin peak duration and amplitude. A beta-adrenergic receptor agonist alone markedly stimulated AA-NAT activity and melatonin synthesis and release. An alpha-adrenergic receptor agonist, while having no effect per se, potentiated the beta-adrenergic stimulation of AA-NAT activity both in vitro and in vivo. Strikingly, the potentiation of AA-NAT activity did not result in a potentiation of melatonin synthesis, suggesting that the rate of melatonin production is limited downstream in the metabolic pathway, most probably at the level of hydroxyindole-O-methyltransferase (HIOMT). HIOMT presented a constitutively high activity that was not acutely (within hours) stimulated by beta-adrenergic agonist, but was rather up-regulated by chronic application of the agonist. This long-term beta-adrenergic regulation may explain the reported large photoperiodic variation of HIOMT activity that drives the photoperiodic variation in melatonin peak.

Acetylserotonin O-Methyltransferase↗