Effect of environmental temperature and photoperiod on the melatonin levels in the pineal, lateral eye, and plasma of the frog, Rana perezi: importance of ocular melatonin

Day-night melatonin concentrations were studied in the pineal body, lateral eye, and plasma of the frog Rana perezi in animals maintained in February and July under long (18L:6D) or short (6L:18D) photoperiod and high (25 +/- 1 degree) or low (6 +/- 1 degree) temperature in order to evaluate the influence of these environmental factors. When frogs were kept under short photoperiod and low temperature in February, no melatonin rhythm was observed in the pineal, ocular tissue, and plasma. High temperature at this period of the year induced a day-night rhythm of melatonin levels in the lateral eye and plasma. In July, under long photoperiod and high temperature, animals showed pronounced rhythms of melatonin in the pineal, eye, and plasma. A decrease of environmental temperature in this season abolished the melatonin rhythm. When animals were maintained in August under high (25 +/- 1 degree) temperature and long (18L:6D) or short (6L:18D) photoperiod, the duration of high night time ocular melatonin levels was correlated to the length of the dark phase. In all experiments the high ocular melatonin concentrations and the close parallelism observed between ocular and circulating melatonin profiles suggest that in this species melatonin could be released from the eyes in the general circulation.     

Effect of Pinealectomy, Eye Enucleation, and Melatonin Treatment on Ovarian Activity and Vitellogenin Levels in the Catfish Exposed to Short Photoperiod or Long Photoperiod

To investigate the relative importance of pineal, and eyes, and melatonin treatment on ovarian activity, catfish. Heteropneustes fossilis subjected to pinealectomy, eye enucleation, or both were exposed to short (LD 9:15) or long (LD 14:10) photoperiod during the different stages of the annual reproductive cycle. During the preparatory period, pinealectomy accelerated ovarian recrudescence under long photoperiod, but no effects of pinealectomy were observed under short photoperiod. Pineal has no influence on ovarian activity during the prespawning and spawning periods of the reproductive cycle. In the late postspawning period, ovarian recrudescence is accelerated after pinealectomy in catfish maintained under short photoperiod (LD 9:15 at 25 degrees C). But under long photoperiod (LD 14:10 at 25 degrees C), pinealectomy delayed ovarian recrudescence. An investigation into the role of the eyes revealed that eye enucleation nullifies the inhibitory effects of LD 9:15 and represses the stimulatory effects of LD 14:10. Combined surgery inhibited ovarian development under both the regimes. It is significant that pinealectomy and/or blinding neither counteracted nor delayed the postspawning ovarian regression. The findings suggest that the role of the pineal in catfish reproduction is variable and depends upon the photoperiod to which they are exposed as well as on the time of the year and the stage of the reproductive cycle. It is concluded that extraretinal and extrapineal photoreceptors also are involved in influencing seasonal reproduction. Further, no effects of melatonin treatment on ovarian activity or on vitellogenin levels during the preparatory or prespawning periods were observed, indicating that the pineal effects on gonadal activity in catfish may not be mediated through the secretion of melatonin.     

Photoperiod influences growth rate and plasma insulin-like growth factor-I levels in juvenile rainbow trout, Oncorhynchus mykiss

The effect of different photoperiod regimes and the subsequent influence of melatonin on growth and insulin-like growth factor-I (IGF-I) were assessed in juvenile rainbow trout. In Experiment 1, triplicate groups of all female underyearling rainbow trout were exposed to one of three photoperiods; simulated natural photoperiod (SNP), constant short-days (LD 8:16), or constant long-days (LD 18:6) from June to December 2000 under ambient water temperatures. Fish exposed to LD 18:6 grew to a significantly heavier mean weight than the other treatments. Regression analysis showed a strong correlation between circulating plasma IGF-I, growth rate and temperature. Furthermore, it was apparent that fish exposed to LD 18:6 expressed significantly higher circulating levels of IGF-I. In a second experiment, duplicate groups of all female yearling trout were exposed to one of three photoperiods; SNP, LD 8:16, or constant light (LL), with sub groups receiving either a slow-release melatonin implant (18 mg), sham implant or left intact (control). LL increased growth rate in controls, reaching a significantly greater weight than SNP or LD 8:16 photoperiods but did not affect circulating IGF-I levels. Melatonin implants reduced growth rate in all photoperiod treatments below that of their respective controls but again did not affect circulating IGF-I levels. No differences in growth rate were found in implanted fish between photoperiods suggesting that a diel cycle of melatonin is necessary for the perception of daylength. These results would indicate that extended photoperiods (LD 18:6) may cause direct photostimulation of growth through up-regulation of IGF-I production. In contrast, in the absence of a changing diel melatonin signal, growth appeared to be maintained by a possible underlying endogenous rhythm, which was phase advanced under LL, as such plasma IGF-I levels simply reflected growth rate rather than photostimulation of the somatotropic axis. Overall, these findings indicate that measuring plasma IGF-I may be a useful tool for studying environmental influences on growth in rainbow trout.     

Environmental control and adrenergic regulation of pineal activity in the diurnal tropical rodent, Arvicanthis ansorgei

Like nocturnal rodents, the diurnal tropical rodent Arvicanthis ansorgei shows a daily rhythm in pineal melatonin content. Seasonal and photoperiodic variations in the biosynthetic activity of the pineal gland: arylalkylamine-N-acetyltransferase (AA-NAT), hydroxyindole-O-methyltransferase (HIOMT) activities and melatonin content were measured in male and female A. ansorgei captured near Samaya, Mali, and kept either under artificial laboratory photoperiods [light-dark (LD) cycles: LD 14:10, LD 12:12 or LD 10:14 or caught in the field in Mali and killed at four different times of the year (January, April, June and November). Under artificial photoperiod, the duration of the nocturnal peak of AA-NAT activity and melatonin content increased with the duration of the dark period while the amplitude did not significantly change. In the field, annual variations in the amplitude of the nocturnal melatonin peak were observed with a maximum in April (highest temperature, low humidity and no grass availability, only seeds) and a minimum in November (high humidity, maximum green grass availability). The variations in the amplitude of the melatonin peak were not correlated with changes in AA-NAT HIOMT activities, suggesting that seasonal variations in the amplitude of the melatonin peak are not driven by these enzymes. Daytime injections of the beta-adrenergic agonist, isoproterenol, stimulated melatonin synthesis in January, April and June, but not in November. The annual differences in the amplitude of the melatonin peak as well as the seasonal differences in the response to an adrenergic stimulation suggest that environmental factors other than photoperiod, such as temperature, humidity and consequent food availability, could be important in the regulation of the annual variations in the pineal biosynthetic activity in this species.     

Seasonal and daily plasma melatonin rhythms and reproduction in Senegal sole kept under natural photoperiod and natural or controlled water temperature

The melatonin daily rhythm provides the organism with photoperiod-related information and represents a mechanism to transduce information concerning time of day. In addition, the duration and amplitude of the nocturnal elevation gives information about duration and thus the time of year. In this study, we investigate the existence of an annual rhythm of plasma melatonin in the Senegal sole. Differences in plasma melatonin levels between fish kept at a controlled temperature (17-20 degrees C) and those exposed to the environmental temperature cycle (11.5-25 degrees C) were also examined throughout the year. Spawning was registered in both groups to determine the time of year in which reproductive rhythms occurred. Our results pointed to the existence of an annual rhythm of plasma melatonin at mid-darkness (MD), with the highest levels (203 +/- 44 pg/mL) observed when water temperature reached 25 degrees C. Water temperature influenced nocturnal, but not diurnal melatonin. Daily melatonin rhythms showed seasonal differences, with higher mean nocturnal levels during the summer solstice (138 +/- 19 pg/mL) and autumn equinox (149 +/- 49 pg/mL). When animals were kept at a constant temperature throughout the year, plasma melatonin levels differed from those observed in fish exposed to the environmental temperature cycle. Regarding the reproductive rhythms, spawning was observed at the end of spring in sole kept under natural temperature conditions, whereas no spawning at all was registered in sole reared at a constant temperature. In short, both photoperiod and temperature affected melatonin production in the Senegal sole, transducing seasonal information and controlling annual reproductive rhythms.     

Roles of melatonin in gonadal maturation of underyearling precocious male masu salmon

Testicular maturation of underyearling precocious male masu salmon (Oncorhynchus masou) is affected by photoperiod. It is accelerated by a short photoperiod (light-dark cycles of 8:16 h; LD 8:16) and delayed by a long photoperiod (LD 16:8). Circulating melatonin levels are high during the night and low during the day:the duration of the nocturnal elevation is longer under a short than under a long photoperiod, suggesting mediation of photoperiodic signals by melatonin. This study examined whether melatonin administration mimics short photoperiodic effects and whether it accelerates the testicular development of underyearling male masu salmon reared under a long photoperiod. Fish were randomly selected in June and were divided into two groups. They were reared under LD 16:8 (lights on 04:00-20:00 h) and fed pellets sprayed with melatonin (0.5 mg melatonin/kg body weight/day) or vehicle once a day at 11:00 h until October. The plasma melatonin profile of the melatonin-treated group was similar to that expected under a short photoperiod. Melatonin treatment had a stimulatory effect on the gonadosomatic index and pituitary gonadotropin (GTH) I contents. Plasma testosterone levels were significantly higher in the melatonin-treated group than in the control group in August. However, spermiation was observed in October in both groups and no significant differences were observed in GTH II contents in the pituitary in the two groups throughout the experiment. These results suggest that mimicking a short photoperiod by melatonin administration stimulated testicular development but did not completely activate the brain-pituitary-gonadal axis in precocious male masu salmon. Thus, melatonin is suggested to be one of the factors that mediates the transduction of photoperiodic information to the brain-pituitary-gonadal axis.     

Keeping track of time under ice and snow in a sub-arctic lake: plasma melatonin rhythms in Arctic charr overwintering under natural conditions

Abstract:  Although photoperiod is considered as a major environmental cue for timing of seasonal events in fish, little is known about the photic information perceived by fish in different aquatic environments. The strongly seasonal Arctic charr, Salvelinus alpinus , reside in lakes covered by thick ice and snow throughout the dark winter in the north. In the present study, we have measured diel changes in their plasma melatonin concentrations from September to June in Lake Storvatnet (70°N), northern Norway. In addition, we have measured the in vitro melatonin production of Arctic charr pineal glands held at experimental light conditions. From September to April a diel profile in plasma melatonin was seen in the charr in Lake Storvatn, with highest concentrations at night. This profile reflected the prevailing above-surface photoperiod, even in February when there were minimal changes in sub-surface irradiance between day and night. In June, plasma melatonin was low throughout the 24-hr cycle, despite there being a marked sub-surface difference in irradiance between night and day. At this time the irradiance in night probably remained above the threshold for suppression of melatonin production. The in vitro experiments revealed no endogenous rhythm in the pineal melatonin secretion, supporting the conclusion that the diel profile seen in the Arctic charr in their natural habitat was driven by ambient photoperiod. In conclusion, the Arctic charr appear to keep track of time even under the extreme conditions of high latitudes during winter, when lakes have thick ice and snow cover.     

A test of the coincidence and duration models of melatonin action in Siberian hamsters: the effects of 1-hr melatonin infusions on testicular development in intact and pinealectomized prepubertal Phodopus sungorus

The pineal hormone melatonin is known to play an important role in mediating photoperiodic messages to the reproductive system in seasonal breeding animals. Our goal was to test, in a single experimental paradigm, two hypotheses that have been forwarded to describe how the circadian rhythm of pineal melatonin transmits photoperiodic information to the reproductive system: 1) induction, i.e., a short-day effect, occurs when secreted melatonin and a circadian rhythm of sensitivity to melatonin coincide in time; 2) induction occurs following exposure to elevated circulating melatonin levels for a prescribed duration. In order to determine the relative validity of these hypotheses, we investigated the testicular maturation response to 1-hr daily infusions of 10, 25, and 50 ng of melatonin in pinealectomized intact and prepubertal Siberian hamsters (Phodopus sungorus). Animals received, beginning on day 15 of life, programmed subcutaneous infusions of melatonin or vehicle at one of five time points (19:00-20:00, 20:00-21:00, 21:00-22:00, 24:00-01:00, and 03:00-04:00 hr) for 15 days. In animals gestated and raised in a long photoperiod (LD16:8 = 16L, where L is the duration of light in hours, and D that of dark), melatonin infusion right after lights off (20:00-21:00 hr) significantly retarded gonadal maturation; this dose was ineffective at other times tested. Doses of 10 and 25 ng melatonin were ineffective at all time points. Identical results were obtained in prepubertal hamsters gestated in a short photoperiod (LD10:14 = 10L) and raised in 16L; these results were independent of the presence or absence of the pineal gland. In animals gestated and raised in 10L, melatonin infusions failed to suppress testicular development beyond that induced by the photoperiod; testicular development was maximally suppressed in all groups. The results of these investigations are best explained under the experimental conditions employed here: 1) the photoperiodic gonadal response in juvenile Siberian hamsters is regulated by the coincidence in time of exogenously administered melatonin with an intrinsic rhythm of sensitivity to melatonin, which, under the constraints imposed by our experimental design, occurred at 20:00-21:00 hr; and 2) the duration of the melatonin signal alone, equal in all groups, cannot explain the results.     

Effects of photoperiod and temperature on the binding of follicle-stimulating hormone (FSH) to testicular preparations and plasma FSH concentration in the Djungarian hamster, Phodopus sungorus

The effects of artificial photoperiod and temperature on testicular FSH binding and plasma FSH levels were studied in adult male Djungarian hamsters. In Exp I, hamsters were transferred to long day (LD) photoperiods (16-h light, 8-h dark) after 8 weeks of adaptation in short day (SD) photoperiods (8-h light, 16-h dark), but the ambient temperature was maintained at 25 C throughout the experiments. A marked increase in the total FSH binding per two testes occurred between 10 and 47 days after transfer to LD, a change that was accompanied by testicular growth. Binding of FSH per testicular weight decreased during the same period. Scatchard plot analyses of the parameters indicated that LD decreased both the concentration of FSH binding sites and the equilibrium dissociation constant (Kd). Plasma FSH levels increased between 10 and 47 days after transfer to LD. In contrast, when hamsters reared under LD for 12 weeks were transferred to SD (Exp II), FSH binding per unit weight basis increased 19 weeks after transfer to SD, but the total binding per two testes decreased markedly. In Exp III, sexually mature male hamsters were subjected to different ambient temperature and photoperiods. There were no significant effects of different ambient temperatures on the testicular weight and testicular FSH binding in animals exposed for 8 weeks to either LD or SD. However, plasma FSH levels of hamsters maintained at 25 C under LD was significantly higher than FSH levels at 7 C under LD. A similar effect of temperature on plasma FSH levels was observed in hamsters under SD. The present study indicates that photoperiod is a more important environmental factor than temperature for the regulation of FSH receptor in the Djungarian hamster.     

Pineal and circulating melatonin rhythms in the box turtle, Terrapene carolina triunguis: effect of photoperiod, light pulse, and environmental temperature

Pineal and circulating melatonin concentrations have been measured throughout the 24-hr cycle in the box turtle, Terrapene carolina triunguis, under different conditions of photoperiod and temperature. An obvious effect of photoperiod on the duration of the night rise of pineal and circulating melatonin is observed; the period of elevated melatonin is 4.30 hr in long photoperiod (18L:6D) and 11.00 hr in short photoperiod (8L:16D). A single pulse of 1 hr illumination beginning 1.30 hr after the onset of darkness, in a 16L:8D cycle, has no effect on pineal or circulating melatonin levels. A clear effect of environmental temperature on the amplitude of the day-night rhythm of melatonin production is observed. A possible role of the pineal of poikilotherms in the transduction of several environmental factors, via the daily pattern of melatonin secretion, is hypothesized.     

Old rats are more sensitive to photoperiodic changes. A study on pineal melatonin

After having previously demonstrated that beta-adrenergic stimulation of melatonin under a standard light:dark (LD) cycle regimen of 12:12 is more effective in young than in old pineal glands, we have now studied how the daylength change LD 18:6 affects pineal melatonin secretion and its regulation by the beta-adrenergic system. Young (10 weeks) and old (22 months) male Wistar rats were synchronized with either a standard LD 12:12 for 4 weeks, or acclimatized under the same LD conditions for 4 weeks, then subjected to a long LD 18:6 photoperiod for 1 week. The rats were sacrificed at three time samplings: 0, 4, and 7h after dark onset (HADO) for LD 12:12 or 0, 2, and 3.5 HADO for LD 18:6. Pineal glands were collected and perifused for 480 min. Isoproterenol (10(-4)M) was infused for 20 min, 4h 10 min after the beginning of the perifusion. Basal levels of melatonin production in the young rats displayed a 1.5-2.5-fold increase compared to those in the old rats. Interestingly, mean basal melatonin levels in old rats under standard LD 12:12 conditions were significantly higher (P<0.05) than mean levels at the same relative dark phase intervals under LD 18:6 conditions. Isoproterenol stimulated melatonin production in both young and old rat pineal glands, regardless of time sampling or photoperiodic conditions. The magnitude of the response to 10(-4)M isoproterenol infusion in old pineals was approximately half that found in young glands (P<0.001), and tended to be higher under LD 12:12, in both young and old rat pineal glands, although no significant difference was found in melatonin response between the two photoperiods (P>0.05). This study shows that basal pineal melatonin levels in old rats are more sensitive to photoperiod changes than in young rats. These results also demonstrate that isoproterenol can stimulate both young and old rat pineal glands irrespective of time or photoperiod and confirm previous findings, showing that the melatonin response to isoproterenol is age-dependent and that pineal gland response to isoproterenol is not photoperiod-dependent, at least under our experimental conditions.     

Influences of season and of temperature, photoperiod, and subcutaneous melatonin infusion on the glomerular filtration rate of ewes

Abstract: Influences of season and of temperature, photoperiod, and subcutaneous melatonin infusion on glomerular filtration rate (GFR) were measured in ewes. There was a seasonal difference of GFR between summer (June-August) and winter (December-February) in Tokyo (35°, 35°N); GFR was significantly ( P < 0.05) higher in summer (4.2 ± 0.3 ml/ min/kg) than in winter (3.0 ± 0.2 ml/min/kg). GFR was measured after exposure to three types of photoperiod, 24L: 0D, 12L: 12D, and 0L: 24D, for 7 to 8 days. The value for GFR obtained at 20°C was significantly lower ( P < 0.05) with 0L: 24D than with the other two photoperiodic conditions. GFR obtained during subcutaneous melatonin infusion (20μg/hr for 16 hr/ day for 7 days) with 24L: 0D conditions was significantly ( P < 0.05) lower (2.5 ± 0.1 ml/min/kg) than without infusion (3.8 ± 0.3 ml/ min/kg) and was about the same as that obtained in animals under 0L: 24D conditions. At 30°C, GFR exhibited no difference between the 3 photoperiodic conditions and was always lower than that found at 20°C. Possible influences of melatonin on GFR are discussed.     

Studies on the Kinetics of Melatonin and N-Acetylserotonin in the Rat at Mid-Light and Mid-Dark

The distribution and disappearance of melatonin and N-acetylserotonin after intravenous injection were studied at mid-light and mid-dark. Male rats were adapted under a photoperiod of 12 h light and 12 h dark cycle for a minimum of 1 week. After the intravenous injection of melatonin (10 micrograms), N-acetylserotonin (10 micrograms), or 3H-melatonin (76 ng), blood samples were collected at 1, 3, 5, 10, 20, 30, 45, 60, 75, 90, and 120 min. The concentrations of indoles in the plasma samples were determined by radioimmunoassay, while the radioactivity of 3H-melatonin was determined by scintillation counting. The distribution of melatonin and N-acetylserotonin followed a two-compartment model, showing an initial distribution phase followed by an elimination phase. The biological half-life of melatonin at mid-light and mid-dark were similar but the volume of distribution (V beta and Vc) and metabolic clearance rate (C1s) were greater at mid-dark than at mid-light. The estimated melatonin secretory rate at mid-dark was fivefold greater than that at mid-light. When 3H-melatonin was used, the half-life was significantly longer. A diurnal difference in the distribution was also evident in the N-acetylserotonin studies. The volume of distribution (V beta and Vc) was greater at mid-dark while the C1s was not significantly faster at mid-dark compared with that at mid-light. However, the estimated N-acetylserotonin secretory rate at mid-dark was eightfold greater than at mid-light.     

Melatonin and maturation pace in female three-spined stickleback, Gasterosteus aculeatus

In many animals sexual maturation is controlled by the photoperiod. In mammals, the photoperiodic message is mediated via melatonin, but it is unclear whether this also applies to fishes. Administration of melatonin via the water on a schedule aimed at mimicking a short nonstimulatory photoperiod cycle has been found not to inhibit maturation in male three-spined sticklebacks, Gasterosteus aculeatus, kept under stimulatory long photoperiods. To study whether melatonin affects maturation pace, adult female sticklebacks kept under stimulatory photoperiodic regimes of Light:Dark (LD) 24:0 h (Exp. 1) or LD 16:8 h (Exp. 2) and 18 degrees were treated with melatonin (0, 20, or 80 microg/L water) via the water for 16 h per day. In addition, females were also kept under a nonstimulatory short photoperiod (LD 8:16). The time at which full maturation was achieved (running roe) was noted and the ovaries of nonovulated fish were studied histologically. Most fish under LD 24:0 and LD 16:8 matured (maturation rates in Exp. 1 and 2, respectively: control, 100 and 86%; low-dose melatonin, 83 and 93%; high-dose melatonin, 90 and 75%), whereas almost all females kept under LD 8:16 remained immature (maturation rates: 0% in Exp. 1 and 3% in Exp. 2). There was no difference in maturation pace, proportion of fish maturing, or relative ovarian weights between controls and melatonin-treated fish kept under LD 16:8. Furthermore, there was no difference in proportion of fish maturing or relative ovarian weights between controls and melatonin-treated fish kept under LD 24:0. However, LD 24:0 controls matured significantly earlier than fish receiving the high melatonin dose. Thus, there was an inhibitory effect of the high melatonin dose on maturation pace under LD 24:0. Nevertheless, this effect was small compared to the inhibitory effect of LD 8:16 treatment, suggesting that at least in this season, the major part of the photoperiodic effects in the stickleback is mediated via mechanisms other than circulating melatonin.     

Circadian rhythms of melatonin secretion from superfused goldfish (Carassius auratus) pineal glands in vitro

A flow-through, whole-organ culture (superfusion) system was developed, and goldfish pineal glands were maintained at 25 degrees under light-dark (LD) 12:12 cycles, reversed LD 12:12 cycles, continuous dark (DD), or continuous light (LL) conditions for 48 hr. Under LD 12:12 and reversed LD 12:12 cycles, superfused pineal glands showed a rhythmic melatonin secretion: Scotophase was associated with high titers and photophase with low titers. The melatonin secretion rhythms persisted for two cycles under DD conditions, whereas nocturnal rises were suppressed under LL conditions. After the transition from LL to DD conditions on the third day, melatonin showed a nocturnal increase. These results indicate that melatonin secretion from the superfused goldfish pineal gland is directly photosensitive and that the goldfish pineal gland harbors a circadian oscillator which generates melatonin secretion rhythms.     

The endogenous rhythm of plasma melatonin and its regulation by light in the highveld mole-rat (Cryptomys hottentotus pretoriae): a microphthalmic, seasonally breeding rodent

Abstract:  The day- and night-time levels of plasma melatonin were measured in adult male and female highveld mole-rats, Cryptomys hottentotus pretoriae . This study aimed to assess whether melatonin secretion in this nocturnal, strictly subterranean but seasonally breeding rodent has a day-night rhythm and whether that rhythm is circadian and can be modified by photoperiod. In experiment 1, a day-night rhythm of plasma melatonin was found in all animals housed on a 12L:12D schedule, with significantly higher concentrations in the dark (D) compared with the light (L) phase. The increment of plasma melatonin concentration at night was the same on days 1 and 2 for animals in the control group and animals transferred to constant dark. The animals transferred to constant light substantially reduced the amplitude of the melatonin rhythm on day 2. This suggests that the endogenous melatonin rhythm in C. h. pretoriae has a circadian pattern, which can be synchronized by photoperiod and inhibited by exposure to light at night. In experiment 2, the concentration of plasma melatonin in animals kept under 14L:10D (long day, LD) conditions differed significantly from animals on 10L:14D (short day, SD). This finding supports the notion that C. h. pretoriae is sensitive to changes in day length.     

Alloxan-Induced Diabetes and the Pineal Gland: Differential Effects on the Levels of Pineal N-Acetylserotonin, Pineal Melatonin, and Serum Melatonin

Effects of alloxan treatment on the levels of pineal melatonin, pineal N-acetylserotonin, and serum melatonin were investigated. Male rats were housed under a photoperiod of 12 h light: 12 h darkness and a temperature of 23 +/- 3 degrees C. Three weeks after alloxan (170 mg/kg) or carrier injection (s.c.), the animals were killed at mid-light (1200 h) and mid-dark (2400 h). Pineal and serum indoles were extracted and quantified by radioimmunoassays. It was found that pineal levels of N-acetylserotonin in the diabetic rats were significantly higher (P less than 0.05) than those of the controls. Conversely, pineal and serum levels of melatonin in the control rats were significantly higher (P less than 0.05) than those of the alloxan-induced diabetics. Our results suggest that alloxan-induced diabetes may decrease pineal melatonin synthesis in rats by reducing the activity of hydroxyindole-O-methyltransferase, resulting in a decrease in pineal melatonin secretion.     

Effects of photic environment on ocular melatonin contents in a labrid teleost,the wrasse Halichoeres tenuispinnis

The wrasse Halichoeres tenuispinnis is a labrid teleost that exhibits robust circadian rhythms in locomotor activity under constant light (LL). This fish buries itself in the bottom sand during the subjective-night, thereby suggesting that behaviorally it adjusts its circadian clock to avoid photoreception. In this study, we determined ocular melatonin contents of the wrasse under various photic environments and used ocular melatonin to indicate photoreception. Under light-dark (LD) cycles, ocular melatonin contents of the wrasse exhibited a daily rhythm, with higher levels during the dark phase than those during the light phase. The duration of nocturnal melatonin elevation was longer under LD 9:15 than under LD 15:9. Acute exposure to 2-h light during the dark phase resulted in a significant decrease in ocular melatonin at mid-dark in an intensity-dependent manner. However, acute exposure to different intensities of light for 2h during the light phase had only a small effect on ocular melatonin contents at mid-day. Under LL, ocular melatonin contents in the wrasse reared with bottom sand present exhibited circadian rhythms and were significantly higher than those with transluscent acryl pellets on the bottom. These results indicate that the ocular melatonin rhythm in the wrasse is driven both by the photic environment and by a circadian clock, and that the wrasse that buries itself in the bottom sand can perceive low intensity of light.     

Influence of temperature and photoperiod on plasma melatonin in the mudpuppy, Necturus maculosus.

A melatonin (MEL) radioimmunoassay employed previously only in mammals was used to estimate plasma MEL in a salamander, Necturus maculosus. Validation procedures included thin-layer chromatography of plasma extracts, parallel inhibition curves of authentic MEL and serially diluted plasma and plasma extracts and quantitative recovery of authentic MEL added to pooled Necturus plasma. A diel cycle of plasma MEL was demonstrated in mudpuppies acclimatized for a minimum of 3 weeks under a 12L:12D photoperiod and 15 +/- 1 degrees and sampled in late March. The MEL cycle persisted under a reversed photoperiod (lights on, 1800 hr), but the amplitude of the MEL peak was diminished, and the peak was more than 180 degrees out of phase with the corresponding peak under a normal photoperiod. In animals acclimated to 5 degrees in mid-June under a 12L:12D photoperiod, the diel cycle of plasma MEL continued, but both midphotophase and midscotophase concentrations were reduced compared with 15 degrees controls. The diel cycle was also present in animals acclimated to 25 degrees, but the decrease in MEL concentrations was less marked than that in 5 degrees animals. Photoperiod is apparently the primary cue for cycles in plasma MEL in Necturus, but the cycle can be influenced by temperature.     

Indoleamines and 5-methoxyindoles in trout pineal organ in vivo: daily changes and influence of photoperiod

This study describes the diel rhythms in several indoleamines, melatonin, and related 5-methoxyindoles in the pineal organ of rainbow trout in vivo. In addition, the effect of different photoperiod conditions was evaluated. Melatonin levels displayed clear daily rhythms in the pineal organ of rainbow trout kept experimentally under long (LD 16:08), neutral (LD 12:12), and short (LD 08:16) photoperiods. Duration of melatonin signal was dependent on the night length of prevailing photoperiod, while peak amplitude was higher when lengthening the photoperiod. Significant daily rhythms in 5-HT content, the precursor of melatonin synthesis, were found in neutral and short photoperiod with increases of the amine content just after the light-dark interphase and decreases in the middle of the night, which were more important under short photoperiod. In contrast, no significant 24-h cyclic variation was found in pineal 5-HT content under long photoperiod. Daily profiles in the content of the main 5-HT oxidative metabolite, the 5-hydroxyindoleacetic acid (5-HIAA), outlined those of the amine precursor. The chronograms of both aminergic compounds contrast with those of 5-hydroxytryptophan content, which displayed a net tendency to increase at night. This study also provides evidence for the existence of daily cyclic changes in the content of 5-methoxytryptamine (5-MT), 5-methoxyindoleacetic acid (5-MIAA), and 5-methoxytryptophol (5-MTOL) in trout pineal organ, which were also dependent on photoperiod. The 24-h profiles in 5-MT content correlated well with those of 5-HT, showing a peak at the first hour of darkness in all photoperiodic conditions, and a decay at midnight only in both neutral and long photoperiods. Similarly, the content of 5-MTOL also displayed high values during the day-night transition in trout kept under neutral and long photoperiods, followed by a slow decay all along the night. Finally, levels of 5-MIAA increased in all photoperiods when lights were turned off, being this nocturnal increase maximal in fish kept under LD 16:08. These results suggest that light-dark cycle modulates daily rhythms in pineal indoles and non-melatonin 5-methoxyindoles by acting mainly through the melatonin synthesis activity, which limits the availability of 5-HT for the oxidative and direct methylation pathways. In addition, it seems that a nocturnally increased synthesis of 5-HT might be a requirement for the optimal formation of melatonin and other 5-methoxyindoles in the pineal organ when trout remain under short photoperiods.