Structure-activity studies of melatonin analogues in prepubertal male rats

Comparison has been made between the activity of the pineal hormone melatonin, and several analogues and metabolites in inhibiting sexual development in a protein-restricted prepubertal rat model. Eleven melatonin analogues or metabolites were tested with the aim of evaluating the model as a test of the hypothesis that melatonin acts as a prohormone and that the ring schism metabolites (kynurenamines) mediate many of the effects attributable to melatonin. Although the hypothesis could not be confirmed, modification of the melatonin structure by lengthening the acrylamide side chain or by replacing the 5 methoxy function with fluorine resulted in loss of biological potency. Modification of the melatonin structure to block the two known points of metabolism resulted in no significant alteration in biological activity. Thus 6-chloromelatonin (blocking 6-hydroxylation) and 2,3-dihydromelatonin (blocking oxidative cleavage of the C2-C3 bond) and 6-chloro-2,3-dihydromelatonin remained biologically active. The metabolic products of brain indoleamine-2,3-dioxygenase, N-acetyl-N2-formyl-5-methoxy kynurenamine (aFoMK) and N-acetyl-5-methoxy kynurenamine (aMK), paradoxically were also biologically active.     

Injections of alpha-melanocyte stimulating hormone affect pineal serotonin, melatonin and N-acetyltransferase activity

To determine if exogenously administered alpha-melanocyte stimulating hormone (alpha-MSH) affects nighttime pineal N-acetyltransferase activity, pineal levels of 5-hydroxytryptophan, serotonin and melatonin, and plasma prolactin levels, adult male hamsters were injected at 1900 hr (lights out 2000-0600 hr) with two doses of the peptide and killed at 0300 hr. The low dose of alpha-MSH (200 ng) produced a significant fall in pineal serotonin, pineal NAT activity and plasma prolactin values. The high dose of the peptide (20 micrograms) increased circulating prolactin titers and pineal serotonin levels and caused a concomitant decrease in pineal melatonin levels.     

Effect of neonatal melatonin administration on sexual development in the rat

In order to study the mechanisms by which melatonin modulates sexual development, 5-day-old female Wistar rats have been treated with a single s.c. injection of melatonin, 3 h before the darkness onset. Criteria for sexual development were the age of vaginal opening and the circulating levels of prolactin, LH, FSH and estradiol. Also, pineal melatonin content was measured. There was a precocious puberty (P less than 0.01) in melatonin-treated rats measured by the age of the vaginal opening. An increase in the number of estrous smears over the whole period studied was observed in melatonin-treated animals as compared to controls. Along with these modifications, there was decrease in pineal melatonin content and serum prolactin levels, on day 21 of life (P less than 0.05), with an increase in both parameters on day 30 of age, in melatonin-treated rats as compared to controls, with no modifications at any other time studied. No differences were detected for serum LH levels considering the whole period studied for both groups. There was a faster decrease in plasma FSH levels with age in melatonin-treated animals than in controls. Serum estradiol levels were decreased in the peripubertal period in melatonin-treated rats as compared to controls. All these data suggest that the modifications induced by neonatal melatonin administration on prolactin, FSH and estradiol could be responsible for the precocious puberty shown in this study.     

A novel melatonin antagonist, N-(2,4-dinitrophenyl)-5-methoxytryptamine neutralizes some effects of melatonin in the female Syrian hamster

In this present study we evaluated the ability of a recently synthesized melatonin antagonist, N-(2,4-dinitrophenyl)-5-methoxytryptamine (ML-23), to antagonize the effects of afternoon injections of melatonin on the reproductive and thyroid axes in the female Syrian hamster. Thirty-six animals were divided into four groups and treated daily for 13 weeks with an afternoon injection of melatonin (25 micrograms/injection) or saline diluent. ML-23 was given via the drinking water to both melatonin- and saline-treated groups. The experiment was continued until 78% of melatonin-treated animals exhibited acyclicity. The results show that ML-23 partially reversed the effects of melatonin on pituitary follicle-stimulating hormone concentrations but was without effect on the decreased pituitary and plasma prolactin concentrations induced by melatonin treatment. Furthermore, ML-23 antagonized the effects of melatonin on plasma thyroxine levels and significantly increased plasma triiodothyronine concentrations and the free triiodothyronine index when used in combination with melatonin. The decrease in ovarian weight and plasma estradiol, but not progesterone, obtained with melatonin treatment also was reversed by ML-23. Our data suggest that ML-23 prevents the effects of melatonin treatment on ovarian weight, pituitary follicle-stimulating hormone levels, plasma estradiol, and thyroxine concentrations in the female Syrian hamster. Since ML-23 did not prevent the effects of melatonin on pituitary weight, plasma luteinizing hormone and prolactin, and pituitary prolactin concentrations, the actions of ML-23 may involve only peripheral sites of action of melatonin. Alternatively, the dose of ML-23 may not have been optimal to prevent all of the central effects of the indoleamine.     

Effects of melatonin treatment on the onset of ovarian activity,reproductive parameters and PRL plasma levels of immature ewes

The aims of this study were to investigate the effect of chronic treatment with melatonin on prolactin plasma profiles, the onset of ovarian activity and the fertility of immature ewes. Beginning in late June, 30 maiden ewes were administered 2.5 mg melatonin (i.m.) daily until mid-September. Progesterone and prolactin plasma concentrations were determined by validated radioimmunoassays on samples collected every 5 days from 17 June until 31 October. Prolactin (PRL) plasma concentrations in the control animals were highest at the beginning of the experiment and lowest towards its conclusion. In melatonin-treated ewes, PRL levels dropped just after treatment began and were similar to those observed in the control animals at the end of the experimental period. The ovarian activity was advanced by approximately one month by the administration of melatonin, while the mean date of lambings was advanced by about two weeks compared with the controls. Fertility in the treated animals was very similar to that of the controls; the prolificacy (lambs/pregnant ewe) was 2 in the treated and 1.62 in the control ewes.     

Effects of shortened daylength and melatonin treatment on plasma prolactin and melatonin levels in pinealectomised and sham-operated ewes

Comparisons have been made between the effects of shortened daylength and melatonin treatment on plasma prolactin and melatonin levels in pinealectomised (Px) and sham-operated (Sh) ewes. Twenty-two anoestrous Merino crossbred ewes, maintained under normal grazing conditions, were assigned to four groups for a period of 9 weeks. Group 1 remained untreated (control), Group 2 was herded into a dark shed at 1600 h each day until dark (approx 4 h), ewes in Group 3 were injected with 100 μg melatonin s.c. at 1600 h each day and ewes in Group 4 were implanted with a melatonin capsule releasing 125–200 μg/day. Another group (Group 5) of 4 Px and 4 Sh ewes from the same flock was maintained in an animal house and subjected to shortened daylength (10. 5 h L : 13. 5 h D, lights off 1600 h). Three weeks after the treatments began, ewes in Groups 1–4 were exposed to a fertile ram and ewes in Group 5 to a vasectomised ram and the day of mating noted. No differences were evident between Groups 1–4 in the ewes' response to the ram, time taken to conceive, duration of gestation or number of lambs born. In untreated Px ewes no plasma melatonin (< 20 pg/ml) was found in either day or night samples, whereas intact animals showed the characteristic night-time rise. The silastic implants produced stable daytime blood levels of 90–120 pg/ml, whereas a single injection of 100 μg melatonin caused a transitory (2–3 h) rise. Shortened daylength (Group 2) or a single daily injection of melatonin (Group 3) lowered prolactin levels but only in ewes with an intact pineal gland, whereas melatonin implants (Group 4) caused a reduction in plasma prolactin in both Px and Sh sheep. The results indicate that light-induced alterations in prolactin production in sheep involve both the pineal gland and melatonin. Continuous melatonin release from implants caused changes in plasma prolactin levels similar to those seen following exposure to short days.     

Studies on the minimal dosage of melatonin required to inhibit pineal antigonadotrophic activity in male golden hamsters.

Blinding adult male golden hamsters was followed by atrophy, within 12 weeks, of the testes and accessory sex organs (seminal vesicles and coagulating glands) and by a significant reduction in pituitary prolactin levels. In experiment 1 blind hamsters received subcutaneously implanted melatonin-beeswax (1:24 mg) pellets at the following intervals: once per week, per 2, 3, 4, 6 weeks, or only one pellet during the 12-week experimental period. The melatonin-beeswax pellets, regardless of the frequency of implantation, overcame completely the inhibitory effects of blinding on reproduction and nearly completely the depressant action of light deprivation on pituitary prolactin levels. In the second study the melatonin-beeswax pellets were implanted subcutaneously into blind hamsters every 2 weeks. The pellets contained either 1 mg, 500, 100, 50, or 1 mug melatonin. With the exception of the 1-mug dosage, melatonin again negated almost totally the inhibitory action of darkness on the gonads and accessory organs and also, for the most part, prevented the drop in pituitary prolactin levels. Based on these studies, when melatonin is chronically administered subcutaneously in a beeswax pellet the minimal dosage of melatonin required to counteract the inhibitory effect of darkness on reproduction seems to be less than 3.6 mug/day. The effects of chronic melatonin treatment are similar to those of pinealectomy.     

Plasma prolactin variations and onset of ovarian activity in lactating anestrous goats given melatonin

This experiment was carried out in order to investigate the effects of melatonin treatment on prolactin plasma levels and the onset of cyclicity in anestrous goats. Fifty lactating goats were treated daily with 2.5 mg melatonin (i.m.) from 6 June to 18 September and fifty goats served as a control. The progesterone and prolactin plasma concentrations were determined by radioimmunoassays on blood samples collected once a week until the end of September. The onset of ovarian activity (monitored by the plasma progesterone determination) was advanced in the melatonin-treated does by about one week compared with the controls. Prolactin plasma levels decreased in the control animals from the highest concentrations observed in June to the lowest near the end of the experiment. In the melatonin-treated goats, the prolactin plasma levels dropped after the beginning of the treatment and remained significantly lower than those of the untreated does for about two months; from then onwards, no differences were observed between the two groups until the end of the experiment.     

Restoration of fertility in light-deprived female hamsters by chronic melatonin treatment

Summary Blinding young adult female hamsters was followed by functional involution of the ovaries and uteri and by the cessation of cyclic vaginal phenomena. Light deprivation was also accompanied by elevated plasma and pituitary levels of luteinizing hormone and depressed levels of prolactin in both the blood and the pituitary gland. Only one of 15 blinded hamsters became pregnant when they were exposed to fertile males for 30 days. Both pinealectomy or chronic melatonin treatment (1 mg melatonin implanted subcutaneously per week in beeswax) prevented the changes in the reproductive organs and in pituitary hormone levels attendant on light-deprivation. Both treatment also returned vaginal cycles to normal and restored plasma prolactin titers. Unlike hamsters that were blinded only, light deprived hamsters that were either pinealectomized or melatonin treated were capable of reproducing when they were caged with fertile males. The reproductive capability (i.e., percent of animals that become pregnant and the sizes of their litters) of these animals was equivalent to that of the untreated control hamsters. This is the first report that chronic melatonin treatment restores fertility in blinded female hamsters.Supported by Grant GB-43233X from the National Science Foundation     

Effects of melatonin and natural and synthetic analogues of arginine vasotocin on plasma prolactin levels in adult male rats

A significant elevation in plasma prolactin was observed 10 min following the intravenous injection of 100 microgram of melatonin into either estrogen-progesterone (EP) primed or into nonsteroid-treated male rats. 60 min postinjection in the EP primed rat, the groups treated with 100 microgram or 10 mg of melatonin had signficantly elevated plasma prolactin levels while no effect was observed with these same doses in the nonsteroid-treated rats. Compared to diluent-treated controls, a significant elevation in plasma prolactin was observed at 10, 20 and 60 min following the intravenous injection of either 1 microgram arginine vasotocin (AVT) or 1 mg melatonin into EP primed male rats. A consistent rise in plasma prolactin was also evident after the injection of 1 microgram of either arginine vasopressin, lysine vasopressin or AVT. Oxytocin had no effect on plasma prolactin values. The intravenous administration of 1 microgram of (deamino-1,6 dicarba, 8-arginine)-vasotocin caused a significant elevation of plasma prolactin 10 and 20 min after injection. However, the injection of another analogue of AVT, (4-leucine, 8-arginine)-vasotocin, had no effect on prolactin release at the time points measured.     

Inhibitory action of exogenous melatonin, 5-methoxytryptamine, and 6-hydroxymelatonin on sexual maturation of male rats: activity of 5-methoxytryptamine might be due to its conversion to melatonin

The effect on sexual maturation of 6 different pineal indoles, including melatonin, and of the metabolite 6-hydroxymelatonin was studied in the male rat after daily injections from 20 to 40 days of age. Only 5-methoxytryptamine (5MT) and 6-hydroxymelatonin (6M), in addition to melatonin, inhibited the neuroendocrine-reproductive axis during sexual maturation. Their potencies when injected in the afternoon were in the range of one-twentieth to one-fifth that of melatonin. Like melatonin these two indoles had no effect when injected in the morning. N-acetylserotonin, serotonin, 5-hydroxytryptophol and 5-methoxytryptophol did not influence sexual maturation either when injected in the morning or in the afternoon. Chromatographic separation was performed on plasma extracts from rats injected daily with the biologically active indoles and killed 10-120 min after the last injection. This procedure confirmed that 6M injections did not increase plasma melatonin levels. In contrast, plasma melatonin levels in 5MT-treated rats were increased 1 h after the 5MT injection. These results suggest that 5MT or part of it might be acetylated to melatonin; thus inhibition of sexual maturation might be mainly due to melatonin. These results indirectly support the contention that melatonin is the principal pineal indoleamine playing a role during sexual maturation.     

Effects of long-term treatment with melatonin or melatonin plus ectopic pituitary transplants on testicular LH/hCG and prolactin receptors in juvenile Syrian hamsters (Mesocricetus auratus)

Juvenile hamsters were injected daily with melatonin and some were also given transplants of 2 pituitaries under the kidney capsule. Weights of the testes and the accessory reproductive glands were reduced after 8 and after 12 weeks of melatonin treatment, but remained unaltered in animals treated with ectopic pituitary transplants. Levels of testicular LH/hCG receptors were significantly reduced by daily melatonin injections for 8 and 12 weeks. The presence of pituitary transplants in melatonin-injected hamsters prevented these reductions, and increased LH/hCG receptors above control levels. These changes in testicular LH/hCG receptors were closely related to alterations in serum prolactin concentration induced by melatonin and pituitary transplants. After 8, but not after 12 weeks of treatment, testicular prolactin receptor levels were reduced by melatonin and maintained by the presence of pituitary transplants. We conclude that: juvenile male hamsters become sensitive to the effects of daily melatonin injections when they reach maturity; daily melatonin injections can reduce the levels of testicular LH/hCG and prolactin receptors; and the effects of melatonin on LH/hCG and prolactin receptors are probably due to suppression of endogenous prolactin release.     

Circadian variations in plasma concentrations of melatonin and prolactin during breeding and non-breeding seasons in yak (Poephagus grunniens L.)

Circadian variations of plasma melatonin and prolactin concentrations were determined during breeding as well as non-breeding seasons in yak. Blood samples (5 ml) were collected during different phases of estrous cycle, viz. early (0-6 days), mid (7-12 days) and late luteal (13-19 days) at 2 h interval for 24 h from eight yaks during one breeding month (November); the same yaks were bled at 2 h interval during one non-breeding month (February) for 24 h. Plasma melatonin concentrations rose sharply (P < 0.01) after sunset to record peak concentrations between midnight and 2 a.m. declining sharply thereafter in both breeding as well as non-breeding seasons. Basal melatonin concentrations were recorded between 0600 and 1600 h. Stage of luteal phase did not influence the diurnal hormone change (P < 0.01). In the breeding season, mean plasma prolactin concentrations displayed circadian variations with maximum value at 0400 h (41.22+ /- 1.5 ng/ml) and minimum at 1400 h (12.0 +/- 4.02 ng/ml). In the non-breeding season plasma prolactin concentrations showed circadian variation with maximum value at 0000 h (59.9 +/- 10.5 ng/ml) and minimum at 1200 h (32.13 +/- 3.2 ng/ml). A positive correlation in breeding (r = 0.75) and in non-breeding season (r = 0.65) between circadian changes in mean plasma prolactin and melatonin concentrations were seen. Circadian changes of mean plasma melatonin concentrations during breeding and non-breeding seasons were not different (P > 0.05). However, mean plasma prolactin concentrations were found to be higher (P < 0.01) in the non-breeding season. Three conclusions were drawn from the study: (i) melatonin and prolactin concentrations followed a circadian pattern of secretion (ii) melatonin and prolactin secretion may be closely interrelated and (iii) higher prolactin concentrations during the non-breeding season could be due to nutritional and environmental stress and hence might be contributing to lack of cyclicity.     

Nocturnal enhancement of plasma melatonin could be suppressed by benzodiazepines in humans

Plasma melatonin levels were determined every 20 and 30 min for 24 hours on the last day of repeated oral administrations (1 or 2 mg a day for 8 or 9 days) of a benzodiazepine derivative (450191-s), which is known to be metabolized to active benzodiazepines after administration. In one of the two subjects, the nocturnal enhancement of plasma melatonin which was obvious on a control day with placebo was diminished almost completely. In the other subject, observed were not only the diminishment of its nocturnal enhancement but also its increase during the daytime almost to the nocturnal levels on a control day, which may indicate a rebound increase in melatonin synthesis or a shift in its day-night rhythmicity. Such suppressing effects of benzodiazepines on the nocturnal plasma melatonin levels were also examined in the case of a single administration of 2 mg of 450191-s or flunitrazepam in the second series of experiments. Even a single flunitrazepam seemed to have lowered nocturnal plasma melatonin levels, which then recovered to the usual levels following the administration of 5 mg of a benzodiazepine antagonist, Ro 15-1788, given 6 hours after the flunitrazepam. However, single 450191-s did not show any remarkable effects. Thus, it has been suggested that benzodiazepines could suppress the nocturnal levels of plasma melatonin or shift its day-night rhythmicity at least when administered repeatedly. The possible action site of benzodiazepines may be the central nervous system, since melatonin synthesis has been though to be under strongly regulated by the central nervous pathway from the retina to the pineal body. Therefore, these effects of benzodiazepines may provide a method for investigating the physiological role of melatonin and its day-night rhythmicity as well as to further clarify the system regulating melatonin synthesis in humans.     

Effects of photoperiod, melatonin implants and castration on molting and on plasma thyroxine, testosterone and prolactin levels in the European badger (Meles meles)

1. The seasonal molt, which lasts six months in the badger, begins in mid-July and ends at the beginning of winter. It occurs under natural long-day conditions, following the seasonal drop in plasma testosterone levels, concomitant with high levels of thyroxine and prolactin. 2. To examine the role of the different factors involved (day length, prolactin, thyroxine, testosterone), different groups of badgers, divided into subgroups of castrated or intact animals, were subjected to the influence of long days (20L: 4D), short days (4L:20D) or the effect of subcutaneous melatonin implants. 3. In all cases, castration resulted in a significantly earlier onset of molting 1-3 months, depending on the group, regardless of the experimental conditions (20L:4D, 4L:20D, melatonin). 4. However, molting started earliest in animals subjected to long days, irrespective of whether they were castrated or intact. 5. In the melatonin-implanted badgers, molting started either early (castrated animals), or late or not at all (intact animals). 6. Lastly, in castrated badgers subjected to experimental photoperiods (short days or long days) or melatonin implants, the period of molting was shortened from 6 months (intact outdoor animals) to 4 months. 7. The advance in shedding was always related to an early drop in testosterone (or an absence of testosterone in the castrated animals) and to a higher or earlier increase in thyroxine levels.(ABSTRACT TRUNCATED AT 250 WORDS)     

Increased REM sleep associated with melatonin deficiency after pinealectomy: a case study

The objectives of the investigation were to assess hypersomnia, which progressively appeared in a young patient after a pinealectomy, chemotherapy, and radiotherapy for a typical germinoma, as well as the potential benefit of melatonin administration in the absence of its endogenous secretion. 24 h ambulatory polysomnography and the Multiple Sleep Latency Test (MSLT) were performed; in addition, daily plasma melatonin, cortisol, growth hormone, prolactin, and rectal temperature profiles were determined before and during melatonin treatment (one 2 mg capsule given nightly at 21:00 h for 4 weeks). MSLT showed abnormal sleep latency and two REM sleep onsets. Nighttime total sleep duration was lengthened, mainly as a consequence of an increased REM sleep duration. These parameters were slightly modified by melatonin replacement. Plasma melatonin levels, which were constantly nil in the basal condition, were increased to supraphysiological values with melatonin treatment. The plasma cortisol profile showed nycthemeral variation within the normal range, and the growth hormone profile showed supplementary diurnal peaks. Melatonin treatment did not modify the secretion of either hormone. The plasma prolactin profile did not display a physiological nocturnal increase in the basal condition; however, it did during melatonin treatment, with the rise coinciding with the nocturnal peak of melatonin concentration. A 24 h temperature rhythm of normal amplitude was persistent, though the mean level was decreased and the rhythm was dampened during melatonin treatment. The role of radiotherapy on the studied parameters cannot be excluded; the findings of this case study suggest that the observed hypersomnia is not the result of melatonin deficiency alone. Overall, melatonin treatment was well tolerated, but the benefit on the sleep abnormality, especially on daytime REM sleep, was minor, requiring the re-introduction of modafinil treatment.     

Seasonal changes in ovulatory activity, plasma prolactin, and melatonin concentrations, in mouflon (Ovis gmelini musimon) and Manchega (Ovis aries) ewes

Seasonal changes in ovulatory activity, plasma prolactin and melatonin concentrations were monitored in a wild (Mouflon) and a domesticated (Manchega) breed of sheep, both originating and living under similar latitudes (40 degrees N). Mouflons express ovarian cycles significantly later than Manchega ewes (October vs. July, P < 0.001); however, they ended cycling one month later than Manchegas (April vs. March, P < 0.05). While prolactin concentrations were high when Manchega ewes started to cycle, they were at their lowest concentrations when Mouflons started cycling. Overall, mean prolactin concentrations were higher (P < 0.001) in Mouflons than Manchegas throughout most of the year. Within the limits of sampling frequency, the duration of melatonin secretion was similar in both groups during the solstice and equinox periods; however, the amplitude was lower (P < 0.01) in Mouflons than Manchegas during the solstice periods. The significant breed differences in the seasonal hormonal changes may be attributed to a genetic influence in the endocrine responses to the same photoperiodic cues.     

The decrease in plasma melatonin at metamorphic climax in Rana catesbeiana (bullfrog) tadpoles is induced by thyroxine.

Melatonin decreases in the plasma of Rana catesbeiana (bullfrog) tadpoles at the climax of metamorphosis when the thyroxine (T(4)) level peaks. Since melatonin inhibited thyroid function in vitro, it would be of interest to determine if the decline in plasma melatonin permits greater thyroid hormone secretion, or if the increasing levels of T(4) cause the climactic decrease in plasma melatonin. The reciprocal effects of administering T(4) or melatonin just prior to metamorphic climax were examined in tadpoles kept at 22 degrees C on an 18L:6D cycle. If melatonin functions as a thyroid antagonist at later metamorphic stages, administration of melatonin should decrease plasma T(4), whereas if T(4) causes the decline in plasma melatonin, T(4) treatment of tadpoles prior to climax should induce the climactic melatonin decrease prematurely. Once daily injection of 40 microg melatonin for 5 days at 19.30 h had no effect on metamorphic progress, or on plasma T(4) or melatonin levels, except for a transient rise in melatonin just after the injection. Immersion in 2.2x10(-4) M melatonin for 6 days accelerated metamorphosis and decreased plasma melatonin, but had no effect on plasma T(4). Administration of T(4) by injection of 0.2 microg, or immersion in a 6.3x10(-8) M solution accelerated metamorphosis more than melatonin immersion, raised plasma T(4) to climax levels, and induced a decrease in plasma melatonin. We conclude that rapid clearance of exogenous melatonin from the circulation in these experiments did not allow it to affect plasma T(4), and that there is clear evidence that the rise in T(4) induces the climax decrease in plasma melatonin. The finding that immersion in a high level of melatonin can lower plasma melatonin and accelerate metamorphosis, whereas a single daily injection does not, provides an explanation for some of the contradictory reports in the literature concerning melatonin's effect on tadpole metamorphic progress.     

Protective effects of melatonin against carbon tetrachloride hepatotoxicity in rats

Melatonin is an indolamine, mainly secreted by the pineal gland into the blood of mammalian species. The potential for protective effects of melatonin on carbon tetrachloride (CCl(4))-induced acute liver injury in rats was investigated in this work. CCl(4) exerts its toxic effects by generation of free radicals; it was intragastrically administered to male Wistar rats (4 g kg(-1) body weight) at 20 h before the animals were decapitated. Melatonin (15 mg kg(-1) body weight) was administered intraperitoneally three times: 30 min before and at 2 and 4 h after CCl(4) injection. Rats injected with CCl(4) alone showed significant lipid and hydropic dystrophy of the liver, massive necrosis of hepatocytes, marked increases in free and conjugated bilirubin levels, elevation of hepatic enzymes (alanine aminotransferase and aspartate aminotransferase) in plasma, as well as NO accumulation in liver and in blood. Melatonin administered at a pharmacological dose diminished the toxic effects of CCl(4). Thus it decreased both the structural and functional injury of hepatocytes and clearly exerted hepatoprotective effects. Melatonin administration also reduced CCl(4)-induced NO generation. These findings suggest that the effect of melatonin on CCl(4)-induced acute liver injury depends on the antioxidant action of melatonin.     

Effects of melatonin implants on plasma concentrations of testosterone, thyroxine and prolactin in the male silver fox (Vulpes vulpes)

Melatonin administration in the form of slow-release implants advanced breeding activity in silver fox males when treatment began in June. Plasma testosterone concentrations were significantly higher in treated animals than in controls from September to November, whereas in February and March they were significantly lower. Plasma prolactin concentrations were significantly reduced immediately following melatonin treatment in June but increased to greater levels than control values and 'peaked' after 7 months. This 'peak' was associated with a rapid decrease in testosterone secretion. The normal seasonal spring rise in prolactin secretion was prevented by melatonin administration. Thyroxine values decreased and were significantly lower after 2 months of melatonin treatment.