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European hamsters (Cricetus cricetus) show pronounced seasonal changes in their physiology and behavior. The present study provides a detailed analysis of the temporal relationship between seasonal cycles of reproduction and body mass and seasonal changes of two circadian parameters, i.e., locomotor activity and 6-sulphatoxymelatonin (aMT6s) excretion, in individual animals kept under natural light conditions. Our results demonstrate a characteristic pattern of locomotor activity and aMT6s excretion observed around the summer solstice, i.e., from mid-May to mid-July. During this time, locomotor activity was characterized by a high level of activity and an early activity onset, while the nightly elevation of melatonin was reduced to baseline levels. These seasonal changes in aMT6s excretion and locomotor activity were only loosely related to changes in the reproductive status of the animals, but correlated well with a period of the annual cycle during which the animals were sensitive to short days. They may therefore reflect a specific state of the circadian pacemaker system within the SCN and can thus be a valuable tool to further characterize molecular and physiological mechanisms of photoperiodic time measurements in European hamsters.

Long term changes in the secretion of FSH, LH and testosterone, and the size of the testis were measured in groups of hypothalamo-pituitary disconnected Soay rams (HPD rams, n = 8) and control Soay rams (HPD-sham operated and unoperated, total n = 8) while exposed to an artificial lighting regimen of alternating 16-weekly periods of long days (16L: 8D) and short days (8L: 16D), and when treated with a constant-release implant of melatonin given under long days (total study: 136 weeks). Short term provocation tests using NMDA (glutamate receptor agonist), GnRH and LH were used to assess functionality of the hypothalamus, pituitary gland and testis, respectively. Control rams expressed normal photoperiod-induced cycles in the reproductive axis. Blood concentrations of FSH, LH and testosterone were significantly increased under short days, and decreased under long days with parallel changes in testicular diameter. Treatment with implants of melatonin under long days mimicked the effect of short days and induced rapid reactivation of the reproductive axis. In the HPD rams the blood concentrations of FSH, LH and testosterone declined immediately after the HPD surgery and values remained close to the lower limit of detection of the radioimmunoassays throughout the experiment. LH pulses were absent in the HPD rams and NMDA failed to induce LH secretion consistent with functional disconnection of the pituitary gland from the hypothalamus. The testes regressed to a significantly smaller size in the HPD rams compared with controls even at the nadir of the sexual cycle (testis diameter: 30.2 +/- 0.7 vs 41.3 +/- 0.8 mm, HPD vs control rams, respectively). A low amplitude cycle in testicular diameter (peak to nadir: 5.0 +/- 0.7 mm) persisted in the HPD rams with a temporal pattern opposite to the controls (growth under long days instead of short days; 'hamster like'). In the HPD rams, the treatment with melatonin blocked the long day-associated increase in testicular size, without effects on FSH, LH and testosterone secretion, pituitary responsiveness to GnRH (LH increment) or testicular responsiveness to LH (testosterone increment). This was in contrast to the cyclical changes in all parameters induced by melatonin in the control rams. At post-mortem, the reproductive tract in HPD rams was markedly regressed compared with the controls. The efficiency of spermatogenesis was reduced with few germ cells maturing beyond primary spermatocytes. Immunocytochemical staining, however, revealed the maintenance of androgen receptor expression in Sertoli cells, pertibular cells and Leydig cells, and steroid activity as measured by 17 alpha-hydroxylase expression in Leydig cells. Overall, the absence of photoperiod-induced changes in gonadotrophin secretion in the HPD rams illustrates the dependence on regulation by the hypothalamus, presumably through the secretion of GnRH. The residual cycle in the size of the testes in the HPD rams was closely correlated with the photoperiod-induced changes in prolactin secretion which persisted in these animals (summary of previous published data included). The combined results support the view that melatonin acts in the hypothalamus to mediate effects of photoperiod on gonadotrophin secretion and in the pituitary gland to mediate effects on prolactin secretion (dual site hypothesis), and that FSH, LH and prolactin act synergistically to regulate the long-term cycle in testicular activity in the ram.

The density of high-affinity melatonin receptors in the pars tuberalis of different long day breeders exhibits a seasonal variation, with high values in spring-summer and low values in autumn-winter. In all the studies, this decrease in receptor density was always associated with both a short photoperiod and sexual inactivity. Moreover, during the hibernation period, the reduction in receptor number was more marked in hypothermic than in normothermic animals. These seasonal changes in melatonin receptors were restricted to the pars tuberalis, which suggests that this structure represents an important target site for mediation of the photoperiodic effect of melatonin on seasonal functions. The mechanism of melatonin action at this level is discussed.

Day-night variations in pineal and/or circulating melatonin and 5-methoxytryptophol (5-ML) concentrations were measured monthly throughout the year in female European hamsters, Cricetus cricetus, maintained under natural conditions. Pronounced seasonal variations in the day-night rhythm of both melatonin and 5-ML were observed. As previously reported for melatonin, the daily rhythm of both methoxyindoles disappeared in spring and early summer, while a clear day-night rhythm occurred in autumn, winter, and early spring. The amplitude of the day-night variations appeared to be maximum from October until January. An inverse relationship existed between the rhythms of melatonin and 5-ML.

Hamsters exposed to short days undergo gonadal collapse followed by recrudescence and insensitivity to the regressive effects of such photoperiods. This refractoriness may be due to exhaustion of the pineal gland or desensitization of its target. Hamsters whose gonads had spontaneously recrudesced were injected with melatonin (25 micrograms per injection) once daily (known to induce regression in intact hamsters) or twice daily (reported to arrest reproduction in pinealectomized hamsters) for 7 weeks. In neither case did refractory hamsters respond to melatonin treatment. The gonads of intact hamsters treated with melatonin for 21 weeks regressed and spontaneously recrudesced along a normal time course. These results indicate that gonadal refractoriness is due to insensitivity of the target tissues of the pineal gland and imply that melantonin participates in photoperiodic regulation of reproduction in the golden hamster.

A progonadal role of the pineal in both adult and juvenile male Djungarian hamsters has been demonstrated which confirms and extends previous reports. In short day-raised hamsters, stimulation of testicular development was achieved by transfer to long days. This stimulatory effect of long days was blocked by pinealectomy. Gonadal stimulation could be elicited in pinealectomized juveniles by timed daily infusions of melatonin. When given for 4 or 6 h daily at a dose rate of 0.83 or 83 ng/h, melatonin infusions stimulated testicular development and body growth. Measurement of plasma FSH and PRL indicated that this pubertal response resulted at least in part from a stimulatory effect of melatonin on the secretion of these hormones. Longer duration infusions (8 or 12 h) did not stimulate testicular development or body growth, and plasma FSH and PRL levels remained low. The efficacy of the short duration infusions did not appear to depend upon the time of day of administration. These findings are in close agreement with those of a previous study which investigated the antigonadal effect of long duration melatonin infusions. The present results demonstrate that melatonin administration can mimic the progonadal effects of the pineal in this species and add further support to the hypothesis that the pineal influences the reproductive system through changes in the duration of daily melatonin production.

Continuous exposure of male hamsters to short day lengths induces testicular regression. This is followed many weeks later by spontaneous recrudescence of the testes with reinitiation of spermatogenesis and function of the accessory sexual glands. Hamsters at this stage of the annual reproductive cycle are refractory to short photoperiods--even continuous darkness will not induce another bout of testicular regression. Animals refractory to short days are also refractory to the pineal hormone melatonin and a number of investigators attribute spontaneous recrudescence and photo and melatonin refractoriness to a developed target cell insensitivity to endogenous melatonin from the pineal. Refractoriness is terminated by exposure to long days for at least 11 weeks. The pineal gland is reported to be essential for this process. We report here the effects of pinealectomy, daily melatonin injections, and constant-release melatonin implants on the ability of male hamsters to recover from the refractory state. In the absence of the pineal gland, refractory male hamsters did not discriminate (count?) 15 weeks of long days to terminate refractoriness. Daily melatonin injections at 1900 h, but not at 1200 h (lights 0600-2000 h) during the 15 weeks of long-day exposure blocked the recovery from refractoriness. Constant-release melatonin implants abolished the animals ability to measure 12 and 15 weeks of long days to terminate refractoriness. These results demonstrate that general target tissue insensitivity to melatonin cannot account for the refractory state in hamsters, that a multiplicity of target tissues may exist for melatonin to account for its varied roles throughout the annual reproductive cycle in hamsters, and that the pineal gland is intimately involved in the animals' ability to measure a prescribed duration of long days to terminate refractoriness.

The pineal melatonin rhythm has been proposed to represent a physiological signal which mediates the effects of photoperiod in a variety of seasonal breeding mammals. Studies in other species have led to the hypothesis that an endogenous circadian mechanism controls pineal melatonin content and can adapt the melatonin rhythm to reflect shifts in the prevailing light-dark schedule. In this report, photoperiodic regulation of the pineal melatonin content was investigated in the male Djungarian hamster, a species that is a long day breeder and photo-periodic from early life. In the first study, a daily pineal melatonin rhythm persisted after 6-8 weeks of constant dark conditions in a specific phase relationship to the onset of wheel running activity. Pineal melatonin levels increased around the time of activity onset, and high values persisted for 14-17 h. In subsequent studies, adult males [previously maintained under long days (16 h of light and 8 h of darkness)] were subjected to one of three conditions: 1) a 3-h advance in the onset of darkness, 2) a delay in the time of light onset, or 3) both alterations in the photoperiod. The acute 3-h advance in onset of the dark period caused an earlier rise in pineal melatonin production by approximately 1.5 h compared to that in controls. In contrast, the falling phase of the pineal melatonin rhythm was not altered by extension of the dark period. Therefore, aspects of the pineal melatonin pattern, i.e. the rising phase and peak duration, reflected changes in the photoperiod, while the falling phase anticipated the onset of the light period predicted by the previous light-dark schedule. These observations support the hypothesis that melatonin production by the pineal is an endogenous circadian rhythm and, as a physiological signal, may convey information about the daylength.

Light deprivation by blinding in female hamsters was followed by a regression of the reproductive organs, an elevation of pituitary LH concentration and a depression of pituitary prolactin levels. Pinealectomy negated almost completely the effects of light deprivation on the neuroendocrine-reproductive axis. Weekly subcutaneous implants of a melatonin:beeswax pellet completely prevented the pineal gland from inhibiting reproductive physiology in blinded hamsters. The findings suggest that melatonin is not pineal antigonadotrophic factor in female golden hamsters. Melatonin implanted hamsters also had higher than normal levels of plasma prolactin.

Pineal glands of male Syrian hamsters stimulated in vivo with isoproterenol (ISO) for 4 h before the onset of darkness showed a 4-h advance in the timing of the nighttime increases in both N-acetyltransferase activity and melatonin levels. When ISO (1 mg/kg) was administered every 2 h to animals kept in light during the night, a significant increase in melatonin synthesis was observed after 4-6 h. The results suggest that the Syrian hamster pineal gland can respond in vivo to continuous beta-adrenergic stimulation, but a lag period of 4-6 h is required before there is an increase in melatonin synthesis.

Plasma levels of thyroxine (T4), triiodothyronine (T3) and growth hormone (GH) as well as the free T4 and free T3 index (FT4I, FT3I) in 20-month-old female Syrian hamsters kept under long daily photoperiod [light:dark (LD) cycles of 14:10] were significantly depressed compared to the values of 2-month-old animals kept under the same photoperiodic conditions. Cholesterol levels in old females were significantly elevated compared to those in young animals. In a subsequent experiment, young (4 month) and old (15 month) female hamsters were either placed in a short photoperiod (10:14 LD) or a 14:10 LD cycle and given daily afternoon injection of either diluent or 25 micrograms melatonin. After 8 weeks, the old females, regardless of treatment or photoperiod, had significantly lower T4 levels and higher cholesterol and triglyceride levels than did young animals. In 14:10 LD, FT4I and FT3I did not differ between young and old animals, possibly because of the reduced age gap in this experiment. Although the FT4I and FT3I in young females after 8 weeks of treatment with 10:14 LD or melatonin were not significant, the old females did demonstrate a significant suppression of their FT4I and FT3I values.

Peripubertal reproductive development of Siberian hamsters is controlled by maternally derived photoperiodic information and the ambient photoperiod present after weaning. Previous experiments suggested that the maternally derived information is transferred during gestation, not during lactation. Development was examined in several photoperiods following manipulation of gestational and lactational photoperiods; development was influenced by the gestational, but not lactational, photoperiod. Second, effects of the gestational photoperiod were observed in young reared in constant light (LL) from Day 15. Depriving the young of ambient photoperiodic information after Day 15 allows a more direct assessment of the signal received from their dams. Finally, melatonin injections to long-day dams, at certain times of day, caused transmission of a short-day signal to young, as evidenced by their development in LL and light-dark cycles. Thus, a maternally derived signal that is dependent on melatonin influences reproductive development of the young during gestation; the maternally directed pattern of development can subsequently be modified by the youngs' own response to ambient photoperiods after weaning.

The age dependency of the time delay preceding a gonadal response to short photoperiod, blinding, and daily evening melatonin injection in young male and female hamsters has been examined to determine why the reproductive system of prepubertal golden hamsters is apparently unaffected by short-day treatment. Anestrous occurred in 50% of the female hamsters during the 10th week after blinding whether or not blinding occurred at 3, 5, 7, or 9 weeks of age. A similar latency was found following initiation of melatonin injections although the variance was greater (50% became anestrous during the 8th, 11th, 9th, and 8th week after initiation at ages of 3, 5, 7, and 9 weeks, respectively). After transfer to short days, however, females were 18 to 22 weeks old before 50% had become anestrous and the latency intervals were 19, 13, 11, and 10 weeks for transfer at 3, 5, 7, and 9 weeks of age, respectively. In males, mean testicular weights decreased to 50% of control values during the 9th and 10th week following blinding regardless of the age at which blinding occurred; the latency following initiation of melatonin injection or transfer to short days ranged between 8 and 11 weeks. These data support the conclusion that prepubertal male and female golden hamsters 3 weeks of age and older respond to light deprivation and melatonin treatment in the same manner as adults, but that the latency of the response is such that sexual maturity is attained before regression occurs. The response of 3-week-old female hamsters to short photoperiods, however, is apparently confounded by factors not governed by the pineal gland.

Male Djungarian hamsters, reared under long (16L/8D) or short (10L/14D) days, were sacrificed at various ages during the day or night, or at night following a 30-min light pulse. The pineal melatonin rhythm matured similarly under long and short days by 20 days of age. The results are discussed in context of the hypothesis that melatonin mediates the photoperiod effects which forestall puberty in short-day reared hamsters.

To determine which parameter of the day/night pattern of pineal melatonin secretion is the critical component signaling daylength information in the Djungarian hamster, we have developed a method for giving timed sc melatonin infusions in pinealectomized juvenile males. When given for 12 h daily, as little as 10 ng melatonin (14 pg/min) consistently induced testicular regression within 12 days. However, 10 ng melatonin infused for 4 or 6 h daily did not inhibit gonadal development. The effects of these infusions on the reproductive system did not depend on the time of day at which melatonin was administered. In complementary experiments, the minimal daily infusion duration and the critical daylength for induction of testicular regression were determined. The critical length of infusion (7-8 h) was in close agreement with the estimated duration of melatonin secretion during the critical scotophase. These findings support the hypothesis that melatonin mediates the pineal-antigonadal effects of short day exposure in the Djungarian hamster. Furthermore, the data strongly suggest that duration is the feature of nighttime melatonin release that is most important for photoperiodic time measurement in this species.

Male golden hamsters were exposed to short photoperiod at either 20 degrees C or 5 degrees C. After 4 weeks a complete gonadal inhibition was observed in animals kept at 5 degrees C while in animals kept at 20 degrees C such an inhibition was much less. No significant difference in the pattern of pineal and plasma melatonin concentrations was observed between hamsters kept at 20 degrees C and 5 degrees C. If in the golden hamster pineal melatonin secretion is implicated in the transduction of the photoperiodic information, it is probably not implicated in the transduction of thermal information.

Djungarian hamsters which did not respond physiologically to short day conditions were injected daily with melatonin. Hamsters responded to this treatment with typical body weight alterations and molt. Therefore, we concluded that the lack of short day adjustments is not based on insensitivity to melatonin in this species. Pretreatment with short days affected the timing of melatonin-induced body weight loss and molt. Hamsters became refractory to melatonin injections earlier for both traits if pretreated with short days. Low body weight level was maintained for a shorter period of time, whereas duration of molt was not affected. These results might indicate differences in the control of melatonin-induced body weight adjustments and molt.

In Siberian hamsters, transference of photoperiodic information from dam to fetus influences pubertal testicular development of the young when reared either in constant light (LL) or postnatal photoperiods of intermediate length (i.e. 14L:10D). The effects of short photoperiods during gestation can be mimicked by administering melatonin to pregnant females. This experiment examined whether there exists a daily pattern of sensitivity to melatonin when it is administered to pineal-intact pregnant females housed on a long photoperiod. Groups of pregnant and lactating females received melatonin at each hour of the day. The young were not treated with exogenous melatonin. At the approximate time of maturation of their endogenous pineal melatonin rhythm (Day 15), the young were placed in LL to suppress pineal melatonin secretion. Young males were killed at 28 days of age. Afternoon (1200 h-2000 h) and late night (0400 h) injections of melatonin into females caused their male young to develop as though gestation occurred on a short photoperiod. Melatonin injections at other times were ineffective. The daily pattern of effectiveness of exogenous melatonin administration to pregnant females resembles that observed in adult males of this and other hamster species and is consistent with the hypothesis that a daily rhythm in sensitivity to melatonin is involved in the transduction of photoperiodic signals.

Pineal melatonin rhythm in golden hamsters was abolished during hibernation. After arousal in darkness, pineal melatonin increased rapidly regardless of whether the arousal was induced during the day or at night. Rapid increase of pineal melatonin after arousal was markedly diminished in animals exposed to light. In hamsters aroused at midnight, the melatonin rhythm in constant darkness ran with the reversed phase relative to hamsters aroused at noon. Since after arousal the melatonin rhythm obviously starts anew from the same phase, we conclude that the circadian pacemaker driving the rhythm might be arrested during hibernation at the day-time phase.

Exposure to short photoperiod or melatonin treatment brings about gonadal regression in Syrian hamsters. The possible influence of these treatments on the sympathetic nervous system (SNS) in these animals was investigated. Male Syrian hamsters were exposed to either long or short photoperiod or subjected to administration of melatonin or its vehicle solution. Exposure of hamsters to 10 weeks of short photoperiod, significantly reduced the noradrenaline (NA) turnover in the heart. Daily administration of melatonin for 8 weeks also resulted in a similar suppression of NA turnover in the heart. Hamsters that were treated with melatonin maintained a lowered metabolic rate as well, at and below thermoneutral temperature. These findings suggest that in a deep hibernator, short photoperiod could suppress the peripheral sympathetic activity and that melatonin may act as the endogenous mediator.

Daily injections of melatonin were found to retard testicular regression in hamsters exposed to LD 10:14, if the injections occurred in the morning (i.e., 0.5 h after lights on), but not if they occurred in the afternoon (i.e., 6 h after lights on). These results indicate that appropriately times injections of melatonin can at least partially block the inhibitory effects of short days on gonadal activity in the photoperiodic hamster.

Since previous studies have shown that an active pineal gland exerts an inhibitory effect on circulating levels of thyroxin in the Syrian hamster, a study was conducted to determine whether the histology and ultrastructure of the thyroid gland supported the conclusions drawn from the hormone data. The ultrastructure of thyroid glands of blinded male Syrian hamsters was compared to that of intact controls kept under a 14L/10D photoperiod, to that of blinded hamsters also pinealectomized, and to that of blinded hamsters receiving 80 micrograms/ml of melatonin in the drinking water. Serum thyroxin (T4) and serum thyrotropin (TSH) concentrations were determined by radioimmunoassay. After 10 weeks serum thyroxin concentrations were less than 50% of controls and concentrations were significantly reduced. EM examination revealed that blinded hamsters had an increased number of follicular cells with flattened epithelium and nondilated endoplasmic reticulum compared to intact controls. In blinded hamsters that were pinealectomized or treated with melatonin in the drinking water, the ultrastructure of the thyroid was not different from controls and serum thyroxin concentrations were restored to near normal. These ultrastructural data support the conclusion that the pineal gland is required to obtain inhibition of the pituitary-thyroid axis in blinded hamsters and that melatonin has a counter-inhibitory effect when administered via the drinking water.

Specific, sensitive and direct radioimmunoassays have been used to determine the daily patterns of 5-methoxytryptophol (ML) and melatonin in the pineal glands of Syrian hamsters kept in different photoperiods: 8 h light: 16 h darkness (8L:16D), 14L:10D and 16L:8D. A rhythm in pineal ML was evident in animals in all the photoperiods, with high daytime levels (641 +/- 35 (S.E.M.) fmol/gland; n = 162) which dropped to 119 +/- 16 fmol/gland (n = 44) 7.1-7.5 h after lights out. The duration of low night-time ML levels was proportional to the length of the dark phase (1.2 h in 16L:8D, 5.4 h in 14L:10D and 8.4 h in 8L:16D). A marked daily rhythm in melatonin was also present in hamsters in the different photoperiods, with daytime levels of 323 +/- 34 fmol/gland (n = 129) and night-time peak concentrations of 3676 +/- 336 fmol/gland (n = 22). The duration of high nocturnal melatonin levels was dependent upon the length of the dark phase (4.1 h in 16L:8D, 4.5 h in 14L:10D and 12.5 h in 8L:16D). Linear regression analysis revealed a statistically significant inverse relationship between pineal ML and melatonin levels in 8L:16D (P less than 0.001), 14L:10D normal (P less than 0.05) and 14L:10D shifted (P less than 0.001) photoperiods. After advancing the lighting schedule by 10 h (14L:10D, lights off at 04.00 h), pineal ML and melatonin rhythms became entrained to the new lighting regimen. The daily rhythms in pineal ML and melatonin in the Syrian hamster thus depend on the prevailing photoperiod, a reciprocal relationship existing between pineal ML and melatonin concentrations.

Twenty-four groups of adult male Djungarian hamsters (Phodopus sungorus sungorus) housed on 16L:8D received daily injections of melatonin, each group at a different hour of the day. Injections (5 micrograms/0.1 ml ethanolic saline 1:10, s.c.) continued for 15 wk, after which the animals were killed and the testes weighed to assess the efficacy of the injections in causing testicular regression. Two periods of melatonin sensitivity were identified. The first was a single time point 5 min before lights on (0455 h). The second was a 5-h period throughout the late afternoon (1600-2100 h; lights out 2100 h). Regression was complete in the 1700- and 1800-h groups only. A few animals failed to respond or regressed very slowly in the 1900- and 2000-h groups, and in the 1600- and 2100-h groups only partial regression was observed after 15 wk of injection. There was no demonstrable effect of exogenous melatonin administration on the endogenous rhythm of pineal melatonin; the rhythm in injected hamsters was of identical duration and amplitude to that in uninjected controls on the same photoperiod.

Daylength (photoperiod) influences the rate of reproductive development in the juveniles of some photoperiodic species. Recent studies show that daylength during the prenatal period is perceived by the fetus and that this perception can profoundly influence postnatal reproductive and somatic development. Using the photoperiodic Djungarian hamster, we assessed the role of the maternal pineal gland and its hormone, melatonin, in this prenatal perception of daylength. Maternal pinealectomy eliminated the influence of prenatal photoperiod on testicular and body weights of male pups, suggesting that a product from the maternal pineal gland communicates daylength to the fetus. Infusion of the pineal hormone melatonin into pinealectomized dams for various durations during gestation mimicked the effect of varying the prenatal photoperiod on both testicular and body weights. These results indicate that pineal melatonin is involved in this novel form of communication from mother to fetus.

The annual cycle of reproductive activity in the Syrian hamster, Mesocricetus auratus, is the result of interaction between seasonal changes in daylength (photoperiodism) and seasonal changes in responsiveness to daylength (seasonality). The present experiment was designed to investigate the role of the pineal gland and its hormone, melatonin, in the alternation of seasonality (scotosensitivity and scotorefractoriness). Male hamsters were maintained on short daylengths (10L:14D) to establish scotorefractoriness, and then they were transferred to long daylengths (14L:10D) for conversion to scotosensitivity (sensitive to short daylengths). Before transfer to long daylengths, some of the hamsters were pinealectomized and others were sham-operated or unoperated. Some of the pinealectomized hamsters received single daily melatonin or saline injections while on long daylengths. After 14 wk on long daylengths, the hamsters were transferred to short daylengths for 10 wk to test for conversion to scotosensitivity. Pinealectomized hamsters were given three daily melatonin injections while on short daylengths. Such treatment is known to promote gonadal regression in scotosensitive but not in scotorefractory hamsters. Examination of testes after the short daylength interval revealed that exposure of nonpinealectomized hamsters to long daylengths had reestablished scotosensitivity (regressed testes). Pinealectomized hamsters that received no melatonin replacement while on long daylengths remained scotorefractory (enlarged testes), whereas those that received single daily injections of melatonin during long daylengths were found to be scotosensitive. These results indicate that a daily pulse of melatonin during expsoure to long daylengths has an important role in reestablishing responsiveness (scotosensitivity) to short daylengths.