Desensitization of pigment granule aggregation in Xenopus leavis melanophores: melatonin degradation rather than receptor down-regulation is responsible.

Xenopus laevis melanophores express a high density (B(max) 1224 fmol/mg protein) of high-affinity (K(d) 37 pm) cell membrane melatonin receptors. Treatment of melanophores with melatonin resulted in a loss of membrane melatonin receptors reaching a maximum (approximately 60%) by 6 h. In addition to receptor loss, a decline in the potency of melatonin to produce pigment aggregation was observed on prolonged treatment. However, the loss of potency (3.8-fold in 24 h and 162-fold in 96 h) was much slower than loss of receptors, and was completely prevented by inclusion of eserine (100 microm), an inhibitor of melatonin deacetylation in the culture medium. Incubation of melanophores with [(3)H]-melatonin showed that eserine prevented metabolism of melatonin to 5-methoxytryptamine. These results indicate that although receptor density does decline on prolonged treatment, this is not responsible for the diminishing melatonin potency, which is entirely due to degradation of melatonin by deacetylation and subsequent deamination in melanophores.

Metabolism of serotonin to N-acetylserotonin, melatonin, and 5-methoxytryptamine in hamster skin culture.

Biotransformation of [3H]serotonin by cultured hamster skin to 3H-metabolites corresponding to N-acetylserotonin (NAS), melatonin, and 5-methoxytryptamine (5-MT) was demonstrated. This process was time-dependent, with the highest production of radioactive NAS and melatonin metabolites after 3 and 5 h of incubation followed by a decrease in the rate of metabolite release into the media. Conversely, the formation of radioactive metabolite corresponding to 5-MT increased gradually during skin culture, reaching the highest level after 24 h of incubation. The production of 3H-metabolites, corresponding to NAS, melatonin, and 5-MT, was stimulated by forskolin with a maximum effect of forskolin at 10 microM concentration. The gas chromatographic/mass spectroscopy analysis of the fraction eluting at the retention time of NAS standard material showed that it contained NAS, further confirming production and release of NAS into the media by hamster skin. Therefore, we conclude that mammalian skin can acetylate serotonin to NAS and postulate that the NAS is further metabolized by the skin to form melatonin which is subsequently transformed to 5-MT.

[Epiphyseal-thyroid interrelations: effect of calcitonin on indole metabolism in health and in the presence of an excess of thyroid hormones]. / Epifizarno-tireoidnye vzaimootnoshenie: vliianie kal'tsitonina na metabolizm indolov v norme i na fone izbytka tireoidnykh gormonov.

A new regularity was established in a study of epiphyseal thyroid interrelationships: the opposite effect of the thyroid hormones (thyroxin and calcitonin) on the formation of methoxyindoles (melatonin, 5-methoxytryptamine) in the epiphysis. Thyroxin was shown to stimulate and calcitonin to inhibit O-methylation of serotonin in the pineal gland, and combined administration of both hormones levelled their action on one another. The inhibitory effect of calcitonin was more marked if a high basal level of melatonin was stimulated by preliminary administration of thyroxin. The authors assume that such a double control of the thyroid is "protective" for the epiphysis, preventing rapid depletion of reserves in case of excessive saturation of the body with thyroid hormones and maintaining its function in case of a sharp rise of the blood level of calcitonin.

Effect of different photoperiods on the diurnal rhythm of 5-methoxytryptamine in the pineal gland of golden hamsters (Mesocricetus auratus).

This study tested the photo-dependency of the rhythmic synthesis of 5-methoxytryptamine (5-MT) in the pineal gland of golden hamsters. After pargyline administration, pineal 5-methoxytryptamine and melatonin were measured by HPLC in male golden hamsters kept under short and long photoperiod. In both photoperiodic regimes, a clear 5-MT rhythm was observed which fitted a sinusoidal function with high values occurring during the daytime and low values occurring during the night-time. The duration of the low night-time levels was clearly proportional to the length of the dark phase. A marked rhythm of melatonin synthesis was also seen with low daytime levels and high night-time values. An inverse relationship between 5-MT and melatonin levels was observed. Thus, after pargyline administration, the rhythms of 5-MT and melatonin in the pineal gland of golden hamsters are photoperiod-dependent and show a reciprocal relationship.

Day-night rhythm of 5-methoxytryptamine biosynthesis in the pineal gland of the golden hamster (Mesocricetus auratus).

5-Methoxytryptamine is a potent agonist of presynaptic 5-hydroxytryptamine autoreceptors modulating serotonin release in the central nervous system. This methoxyindole can be synthesized in the pineal gland, but its presence in vivo is still controversial, probably because of rapid catabolism by monoamine oxidase. An improved high-pressure liquid chromatography method, with coulometric detection, has been developed for the simultaneous measurement of melatonin, 5-methoxytryptamine, 5-methoxytryptophol and 5-methoxyindolacetic acid. We have demonstrated a day-night rhythmicity in the amount of 5-methoxytryptamine in the pineal gland of golden hamsters (Mesocricetus auratus) maintained under a long photoperiod (14 h light: 10 h darkness) and pretreated with the monoamine oxidase inhibitor pargyline. Levels of 5-methoxytryptamine were highest at 16.30 h and lowest at 00.30 h. The rhythm for 5-methoxytryptamine appears to be the same as for serotonin (opposite in phase to that of melatonin). The identification of 5-methoxytryptamine has been confirmed by analysis with gas chromatography-mass spectrometry.

Rhythmic synthesis of various 5-methoxyindoles in the pineal gland of male adult golden hamsters, kept under the same artificial conditions throughout the year.

Until now the day/night and seasonal rhythmicity in the synthesis of 5-methoxyindoles (MI) is thought to be regulated by environmental factors, especially photoperiod and temperature. Endogenous factors are also implicated in the generation of N-acetyltransferase and hydroxyindole-O-methyltransferase activity rhythms. In the present experiments seasonal rhythmicity in the synthesis of MI in the pineal gland was investigated in hamsters kept under the same artificial conditions throughout the year. Though the environmental conditions were the same, day/night and seasonal rhythmicity in the production of MI in the pineal were observed indicating the existence of endogenous factors influencing the rhythmicities. In November, most of the MI showed the highest synthesis, MA and ML excepted, which were especially produced in July and September. The results obtained sustain the hypothesis that aMT is synthesized from MT rather than from aHT. Moreover, the rhythmicities in aMT synthesis are not identical to those found in aMT concentration as described in the literature. This indicates that synthesis and concentration of a compound are not comparable. At the end of the light period, when aMT injections have an antigonadotropic effect, a peak of aMT synthesis was always present. Although MI synthesis showed seasonal rhythmicity, no reproductive cycle occurred in the hamsters. At present, the concept that the pro- and/or antigonadal effects of the pineal are mediated by aMT seems to be the most acceptable. The present results, however, indicate that aMT and perhaps other MI, often regarded as factors influencing gonadal growth in golden hamsters, are not the only factors involved.

Circadian information and messages in the modified photoreceptor cells of the avian pineal organ: retrospect and prospect.

The modified photoreceptor cell (MP), homologous to the cone-like photoreceptor, is the principal cell type of the avian pineal organ. Two types of secretions (indoles and specific proteins-- pinealins ) are produced in this polarized cell. The synthesis of indoles undergoes striking circadian and circannual changes. MP appear to be receptoneuroendocrine cells which transduce circadian information into 5- methoxyindolic (melatonin, 5-methoxytryptophol, 5-methoxytryptamine) and possibly peptidic neurohormones. Taking into account different stages of differentiation and/or different physiological states (due to cyclic changes) for MP, several mechanisms of transduction of photoperiodic inputs into circadian hormonal outputs have been hypothesized.

The influence of some pteridines on pineal 5-methoxyindole synthesis in male Wistar rats periodically exposed to either white or green light.

The pineals of 28 days old male Wistar rats, in December periodically exposed to either white or green light, were incubated with pterin-6-aldehyde or reduced neopterin. In white light the rhythm of synthesis of 5-methoxytryptophan and of 5-methoxyindole-3-acetic acid was clearly influenced by the pteridines mentioned. In green light a change in rhythmicity of 5-methoxytryptophan, 5-methoxytryptamine and of melatonin/5-methoxytryptophol synthesis by the pteridines was observed. In white light both pteridines increased 5-methoxytryptophan and decreased 5-methoxyindole-3-acetic acid synthesis. Reduced neopterin stimulated 5-methoxytryptamine synthesis and inhibited melatonin/5-methoxytryptophol synthesis. Pterin-6-aldehyde showed an opposite effect. In green light both pteridines decreased 5-methoxytryptophan synthesis, but increased 5-methoxyindole-3-acetic acid and 5-methoxytryptamine synthesis. Melatonin/5-methoxytryptophol synthesis was decreased by reduced neopterin and increased by pterin-6-aldehyde. The results suggest an indolic metabolic pathway leading from 5-methoxytryptophan via 5-methoxytryptamine to melatonin, while pteridines and light of different wave lengths are correlated in regulating indole metabolism.

The pineal gland of the mole (Talpa europaea L.). VII. Activity of hydroxyindole-O-methyltransferase (HIOMT) in the formation of 5-methoxytryptophan, 5-methoxytryptamine, 5-methoxyindole-3-acetic acid, 5-methoxytryptophol and melantonin in the eyes and the pineal gland.

Using a method in which no substrate is added to the incubation medium, the capacity of HIOMT to synthesize 5-methoxytryptophan, 5-methoxytryptamine, 5-methoxyindole-3-acetic acid, 5-methoxytryptophol and melatonin has been determined in the pineal and the eyes of the mole, a mammal having an atrophied visual system. The results demonstrate that the indoleamine metabolism in the retina is similar to the indoleamine metabolism in the pineal. Moreover, in all animals studied both eyes together synthesize 2 to 30 times more of 5-methoxyindoles than the pineal, a result which proves again that the pineal is not the only and not always the most important source of 5-methoxyindoles. With the exception of 5-methoxyindole-3-acetic acid, the synthesis of which is practically neglectable, the production of all other 5-methoxyindoles in the pineal as well as in the retinae is always larger than that of melatonin. In the pineal, 5-methoxytryptophan, for example, is synthesized in a quantity which is 60 to 170 times larger than that of melatonin, while in the retinae the synthesized amount of 5-methoxytryptophan is even 60 to 1000 times larger than that of melatonin.

Seasonal variations in HIOMT activity during the night in the pineal gland of 21 day old male Wistar rats.

In the pineal of 21-day old male Wistar rats hydroxyindole-O-methyltransferase (HIOMT) activities involved in the synthesis of several 5-methoxyindoles were determined during the night in April, June, October and January. A high HIOMT activity for the synthesis of melatonin/5-methoxytryptophol was determined in the months of January and April. In June and October a decrease was observed. The activity maxima coincide with peaks of activity found for the synthesis of 5-methoxytryptophan. Synthesis of 5-methoxytryptamine occurred only in June and October, whereas the synthesis of 5-methoxyindole-3-acetic acid occurred only in January. From these results it may be concluded that January and April are the most active months of those tested for the melatonin/5-methoxytryptophol synthesis in the rat pineal gland. A possible physiological role of the 5-methoxyindoles other than melatonin and 5-methoxytryptophol is discussed.

Turnover of 3H-5-hydroxytryptamine to 3H-5-hydroxyindoleacetic acid and the 3H-5-methoxyindoles in nondeprived and 24 hr food deprived rats.

Serotonin turnover in the lateral in hypothalamus (LH) was determined in nondeprived and 24 hr food deprived rats. The LH was infused with 0.5 muCi of 3H-5-hydroxytrptamine 1 hr prior to push-pull perfusion. The percentage of nCi/muCi of radioactivity was analyzed by thin layer chromatography and liquid scintillation spectrometry. There was significantly more 5-hydroxyindoleacetic acid and 5-methoxytryptamine formed in the 24 hr food deprived rats. These results indicate a faster 5-hydroxytryptamine turnover rate in the LH of 24 hr food deprived rats than in nondeprived rats.