Daily patterns of mRNA expression of two core circadian regulatory proteins, Clock2 and Per1, and two appetite-regulating peptides, OX and NPY, in goldfish (Carassius auratus)

The aim of this study was to examine the daily hypothalamic mRNA expression profiles for two core circadian regulatory proteins, CLOCK2 and PER1, and for two neuropeptides that regulate wakefulness and food intake, OX and NPY, in goldfish. The profiles were determined for fish at different nutritional states (i.e. fed or unfed on sampling day) and held at different photoperiods (i.e. 16L:8D photoperiod vs. constant light LL). Our results show that under a 16L:8D photoperiod, both fed and unfed goldfish exhibit clear antiphasic daily rhythms of hypothalamic Clock2 and Per1 mRNA expression levels, whereas under LL, daily Clock2 rhythms are seen in both fed and unfed fish while significant rhythms of Per1 mRNA expression only persist in unfed fish. In fish held under 16L:8D, but not under LL, there was significantly higher Per1 expression in fed fish at feeding time than in unfed fish. Daily variations in hypothalamic OX mRNA expression levels with peaks observed prior to both feeding time and the onset of darkness, were displayed under a 16L:8D photoperiod, whereas exposure to LL resulted in lower expression levels with no significant daily variations. Fish held under LL, but not under 16L:8D, showed significant daily variations in NPY mRNA expression with a peak prior to feeding time. Taken together, our results suggest that the mRNA expression of both appetite-regulating and circadian proteins display daily variations and that these patterns can be affected by external cues such as feeding and photoperiod.     

The daily melatonin pattern in Djungarian hamsters depends on the circadian phenotype

Djungarian hamsters (Phodopus sungorus) bred at the Institute of Halle reveal three different circadian phenotypes. The wild type (WT) shows normal locomotor activity patterns, whereas in hamsters of the DAO (delayed activity onset) type, the activity onset is continuously delayed. Since the activity offset in those hamsters remains coupled to "light-on," the activity time becomes compressed. Hamsters of the AR (arrhythmic) type are episodically active throughout the 24 h. Previous studies showed that a disturbed interaction of the circadian system with the light-dark (LD) cycle contributes to the phenomenon observed in DAO hamsters. To gain better insight into the underlying mechanisms, the authors investigated the daily melatonin rhythm, as it is a reliable marker of the circadian clock. Hamsters were kept individually under standardized laboratory conditions (LD 14:10, T=22°C±2°C, food and water ad libitum). WT, DAO (with exactly 5 h delay of activity onset), and AR hamsters were used for pineal melatonin and urinary 6-sulfatoxymelatonin (aMT6s) measurement. Pineal melatonin content was determined at 3 time points: 4 h after "light-off" [D+4], 1 h before "light-on" [L-1], and 1h after "light-on" [L+1]). The 24-h profile of melatonin secretion was investigated by transferring the animals to metabolic cages for 27?h to collect urine at 3-h intervals for aMT6s analysis. WT hamsters showed high pineal melatonin content during the dark time (D+4, L-1), which significantly decreased at the beginning of the light period (L+1). In contrast, DAO hamsters displayed low melatonin levels during the part of the dark period when animals were still resting (D+4). At the end of the dark period (L-1), melatonin content increased significantly and declined again when light was switched on (L+1). AR hamsters showed low melatonin levels, comparable to daytime values, at all 3 time points. The results were confirmed by aMT6s data. WT hamsters showed a marked circadian pattern of aMT6s excretion. The concentration started to increase 3?h after "light-off" and reached daytime values 5 h after "light-on." In DAO hamsters, in contrast, aMT6s excretion started about 6?h later and reached significantly lower levels compared to WT hamsters. In AR animals, aMT6s excretion was low at all times. The results clearly indicate the rhythm of melatonin secretion in DAO hamsters is delayed in accord with their delayed activity onset, whereas AR hamsters display no melatonin rhythm at all. Since the regulatory pathways for the rhythms of locomotor activity and melatonin synthesis (which are downstream from the suprachiasmatic nucleus [SCN]) are different but obviously convey the same signal, we conclude that the origin of the phenomenon observed in DAO hamsters must be located upstream of the SCN, or in the SCN itself.     

Pineal melatonin rhythms in female Turkish hamsters: effects of photoperiod and hibernation

Daily rhythms of pineal and serum melatonin content were characterized for adult female Turkish hamsters (Mesocricetus brandti) exposed to long days (16L:8D, 22 degrees C) or after transfer to short days (10L:14D, 22 degrees C). The nocturnal peak of pineal melatonin content was found to be approximately 3 b greater in duration on short than on long days. Changes in levels of serum melatonin closely paralleled those of pineal melatonin. Thus, an effect of photoperiod on synthesis and secretion of pineal melatonin was demonstrated. In a separate experiment, female hamsters were induced to hibernate by exposure to a short-day, cold environment (10L:14D, 6 degrees C). During the 4 to 5-mo hibernation season, Turkish hamsters are known to display 4 to 8-day hours of torpor (body temperature = 7-9 degrees C) alternating with 1 to 3-day intervals of euthermia (body temperature = 35-37 degrees C). Little evidence of nocturnal synthesis or secretion of pineal melatonin was detected in females sampled during torpor. However, animals sampled during the first day after arousal from a torpor bout displayed melatonin rhythms no different in phase or amplitude from those seen in females held at 22 degrees C. Thus, despite the absence of pineal melatonin output during torpor, the pineal gland of hibernating Turkish hamsters produces an appropriately phased, rhythmic melatonin signal during intervals of euthermia.     

Daily scheduling of the golden spiny mouse under photoperiodic and social cues.

A most important function of the circadian system is to ensure that behaviors and metabolism are appropriately timed with respect to the light/dark cycle and photoperiod. Ecological constraints can perturb the daily schedules; would they also impair photoperiodic adaptations? A natural model exists in the golden spiny mouse (Acomys russatus), which is nocturnal, but driven into diurnal activity when sharing the habitat with its congener, A. cahirinus. We show here that the presence of A. cahirinus alters the diurnal rhythms of body temperature and urine volume, delays excretion of the major melatonin metabolite, 6-sulfatoxymelatonin (6-SMT), and increases 2-deoxyglucose uptake by the suprachiasmatic nuclei in A. russatus. Nevertheless, a clear photoperiod effect on urine volume and 6-SMT rhythms was observed. These results indicate that the circadian system can adapt to major changes in daily scheduling without impairing daylength measurement, and consequently seasonal adaptation.     

The thalamic intergeniculate leaflet mediates locomotor activity-induced reversal of phenotype in photoperiod nonresponsive Siberian hamsters

The role of the intergeniculate leaflet of the thalamus (IGL) in photoperiod responsiveness was examined in a laboratory-selected line of photoperiod nonresponsive (NR) Siberian hamsters. NR hamsters fail to exhibit typical winter-type responses (i.e., gonadal regression and development of winter-type pelage) when exposed to short day lengths (e.g., 10 h of light/day). Earlier studies revealed that NR hamsters will exhibit winter-type responses when exposed to short photoperiod if they are given free access to a running wheel. The present study tested the hypothesis that this locomotor activity-induced reversal of phenotype is dependent on the IGL. Male NR hamsters underwent destruction of the IGL prior to being housed in short day lengths in cages equipped with running wheels. Activity rhythms were monitored for 8 weeks, after which time pelage response and paired testes weights were obtained. In contrast to sham-operated NR animals given access to running wheels, IGL-ablated animals showed no increase in the duration of nocturnal running wheel activity and became active later in the night than sham-lesioned animals. Lesioned animals also failed to exhibit the typical short photoperiod-induced gonadal regression and pelage molt. The results implicate the IGL in the mechanism by which running wheel activity can influence photoperiodic responses.     

The Daily Melatonin Pattern in Djungarian Hamsters Depends on the Circadian Phenotype

Djungarian hamsters (Phodopus sungorus) bred at the Institute of Halle reveal three different circadian phenotypes. The wild type (WT) shows normal locomotor activity patterns, whereas in hamsters of the DAO (delayed activity onset) type, the activity onset is continuously delayed. Since the activity offset in those hamsters remains coupled to “light-on,” the activity time becomes compressed. Hamsters of the AR (arrhythmic) type are episodically active throughout the 24?h. Previous studies showed that a disturbed interaction of the circadian system with the light-dark (LD) cycle contributes to the phenomenon observed in DAO hamsters. To gain better insight into the underlying mechanisms, the authors investigated the daily melatonin rhythm, as it is a reliable marker of the circadian clock. Hamsters were kept individually under standardized laboratory conditions (LD 14:10, T?=?22°C?±?2°C, food and water ad libitum). WT, DAO (with exactly 5?h delay of activity onset), and AR hamsters were used for pineal melatonin and urinary 6-sulfatoxymelatonin (aMT6s) measurement. Pineal melatonin content was determined at 3 time points: 4?h after “light-off” [D?+?4], 1?h before “light-on” [L???1], and 1?h after “light-on” [L?+?1]). The 24-h profile of melatonin secretion was investigated by transferring the animals to metabolic cages for 27?h to collect urine at 3-h intervals for aMT6s analysis. WT hamsters showed high pineal melatonin content during the dark time (D?+?4, L???1), which significantly decreased at the beginning of the light period (L?+?1). In contrast, DAO hamsters displayed low melatonin levels during the part of the dark period when animals were still resting (D?+?4). At the end of the dark period (L???1), melatonin content increased significantly and declined again when light was switched on (L?+?1). AR hamsters showed low melatonin levels, comparable to daytime values, at all 3 time points. The results were confirmed by aMT6s data. WT hamsters showed a marked circadian pattern of aMT6s excretion. The concentration started to increase 3?h after “light-off” and reached daytime values 5?h after “light-on.” In DAO hamsters, in contrast, aMT6s excretion started about 6?h later and reached significantly lower levels compared to WT hamsters. In AR animals, aMT6s excretion was low at all times. The results clearly indicate the rhythm of melatonin secretion in DAO hamsters is delayed in accord with their delayed activity onset, whereas AR hamsters display no melatonin rhythm at all. Since the regulatory pathways for the rhythms of locomotor activity and melatonin synthesis (which are downstream from the suprachiasmatic nucleus [SCN]) are different but obviously convey the same signal, we conclude that the origin of the phenomenon observed in DAO hamsters must be located upstream of the SCN, or in the SCN itself. (Author correspondence: weinert@zoologie.uni-halle.de)     

Body temperature and behaviour of golden hamsters (Mesocricetus auratus) and ground squirrels (Spermophilus tridecemlineatus) in a thermal gradient

Golden hamsters and thirteen-lined ground squirrels were maintained individually in a thermal gradient (14°C to 33°C) for several weeks under a 14L: 10D light-dark cycle. Animals of both species showed robust daily rhythms of body temperature and locomotor activity with acrophases consistent with the habits of the species (diurnal acrophases in the diurnal squirrels and nocturnal acrophases in the nocturnal hamsters). Hamsters showed a robust daily rhythm of temperature selection 180° out of phase with the rhythms of body temperature and locomotor activity. Squirrels did not show a daily rhythm of temperature selection. These results raise the hypothesis that a daily rhythm of temperature selection is exhibited by nocturnal but not by diurnal endotherms.     

Effect of chronic hyperprolactinemia on daily changes of glutamate and aspartate concentrations in the median eminence and different hypothalamic areas of male rats

The 24h changes of glutamate (GLU) and aspartate (ASP) were studied in the median eminence (ME) and hypothalamic areas. It was analyzed whether prolactin may change their daily patterns. The hypothalamic concentration of these amino acids was measured by high-performance liquid chromatography (HPLC) with fluorometric detection. Plasma prolactin levels increased over the 24h light-dark cycle after pituitary grafting compared to controls, and its circadian rhythm was disrupted. In controls, aspartate and glutamate in the hypothalamic areas studied followed a specific daily variation or showed no rhythmicity. In the median eminence, hyperprolactinemia seem to phase advance the aspartate or glutamate peaks from 16:00 to 12:00. In the mediobasal hypothalamus, hyperprolactinemia altered daily changes of aspartate and significantly decreased its concentration. Also, it seems to delay the nocturnal glutamate peak compared to controls. In the posterior hypothalamus, hyperprolactinemia did not change aspartate and glutamate concentrations and their daily changes, although it increased the glutamine concentration. These data show the existence of 24h changes of amino acid concentration in three of the hypothalamic regions studied. Increased plasma prolactin levels differentially affected these patterns depending on the hypothalamic area analyzed. (Chronobiology International, 17(5), 631-643, 2000)     

60 Hz magnetic field exposure and urinary 6-sulphatoxymelatonin levels in the rat

Four separate experiments were carried out to investigate the effect of extremely low frequency magnetic field (MF) exposure (60 Hz, 1 mT rms) on urinary 6-sulphatoxymelatonin (aMT6s) levels in Sprague-Dawley rats. In the first experiment, immature male rats maintained under a regular 12 h daily photoperiod (white fluorescent light) were exposed to a 20 h daily MF exposure for 6 weeks. The second experiment was similar to the first, except that the MF exposure was limited to 10 days. In the third experiment, adult male rats acclimated to a combination of continuous dim red light and regular 12 h daily photoperiod (white fluorescent) were subjected to a single 1 h exposure to intermittent MF (1 min on and 1 min off cycles), 2 h before fluorescent lights went off. The fourth experiment was similar to the third, except that the animals received 2 consecutive days of 20 h daily exposure to intermittent MF, beginning 1 h before the fluorescent lights went off each day. In all four experiments, the circadian profile of urinary aMT6s was examined before, during, and after the MF exposure. No significant effect of 1 mT MF on indoleamine metabolism was observed in any of the above experiments. However, in one of the experiments (no. 4), both the control and the MF groups showed a lower aMT6s level during the exposure days, when compared with that of pre- and post-exposure days, suggesting that the existence of possible effects with lower field strengths at the range of stray field cannot be ruled out. Bioelectromagnetics 19:172–180, 1998. © 1998 Wiley-Liss, Inc.     

Search for seasonal rhythmicity of pineal melatonin production in rats under constant laboratory conditions: spectral chronobiological analysis, and relation to solar and geomagnetic variables

Earlier we reported that in a number of experiments pineal melatonin production in rats under constant laboratory conditions displayed seasonal rhythms but subsequently were not always able to confirm this. Since there was no indication under which conditions such rhythms may be present, we performed four consecutive identical experiments with untreated female Sprague-Dawley rats within the same animal room during 1997-2006. Nocturnal urine samples (19-23, 23-3, 3-7?h) were collected at monthly intervals over 494-658?d with 12 animals each in experiments I and II (1997-1999, 1999-2000), 30 animals in experiment III (2002-2004), and 15 in experiment IV (2005-2006). 6-Sulfatoxymelatonin (aMT6s) was measured by ELISA. The excreted aMT6s at each time interval as well as total nocturnal aMT6s-excretion (19-7?h) was submitted to standard statistical analyses as well as to a spectral chronobiological analysis to determine the period lengths of the components involved which was followed by processing with the single cosinor method. Seasonal rhythm components (circannual period length: 360 ± 60?d) were detected in experiment III (2002-2004) for the overall nocturnal excretion as well as for two sub-intervals (23-3 and 3-7?h) and in one night interval of experiment II (23-3?h). Multiple components with mostly short period lengths of around 100?d and some long ones of 500-650?d were found in the other experiments. Systematic MESOR and amplitude variations were observed during the experiments, being highest in experiment II (19-7?h, also 23-3?h and 3-7?h) and lowest in experiments I and IV. These results illustrate that seasonal melatonin rhythms are not a general phenomenon in female laboratory rats indicating an involvement of unknown environmental cues. As an extension of our earlier hypothesis regarding a seasonal Zeitgeber function of the horizontal intensity H of the geomagnetic field showing circannual variations, we assume further modulation by the 11-yrs' sunspot cycle which leads to geomagnetic disturbances and could facilitate seasonal aMT6s rhythmicity during specific years. (Author correspondence: christian.bartsch@uni-tuebingen.de ).     

Synchronizing Effect of Photoperiod on the Dual Phasing of Demand-Feeding Rhythms in Sea Bass

Under natural environmental conditions, sea bass feeding rhythms are nocturnal in winter and diurnal the rest of the year. In this paper we describe the effect of contracting and expanding photoperiods and two skeleton photoperiods (SP) on the dual feeding rhythms of sea bass ( Dicentrarchus labrax L. ). To this end, twelve animals were held individually with access to self-feeders. First, the lights on and lights off were progressively delayed and advanced respectively by one hour in group 1 (G1), and conversely in group 2 (G2), so that the fish were exposed from a light/dark (LD) 12L:12D cycle to 2:22 LD (G1) and DL (G2) cycles and finally 0.25:23.75 LD (G1) and DL (G2). In the second experiment two SP's were used involving two light pulses separated by 12 hours, each pulse lasting 0.25 hours during the first two weeks and one hour during the succeeding two weeks. The results showed that diurnal and nocturnal sea bass tended to confine their feeding phase following the contraction of the LD cycle. Both SP's failed to simulate a complete photoperiod. In conclusion, the LD cycle appeared to drive the daily feeding rhythms but, the photoperiod length did not itself control the inversions of nocturnal and diurnal fish, so that other factors, in addition to photoperiod, may be involved in the control of the annual rhythms of phase inversions in sea bass.     

Daily Rhythms of Metabolic Rate and Body Temperature of Two Murids from Extremely Different Habitats

Daily circadian rhythms of body temperature (T_b) and oxygen consumption (VO₂) were measured in two murid species, which occupy extremely different habitats in Israel. The golden spiny mouse (Acomys mssalus) is a diurnal murid distributed in arid and hot parts of the great Syrio-African Rift Valley, while the broad-toothed field mouse (Apodeinns mystacinus) is a nocturnal species that inhabits the Mediterranean woodlands. In both species, the daily rhythms of T_b and VO₂ are entrained by the photoperiod. Under laboratory experimental conditions (ambient temperature T_a = 33^oC and photoperiod regime of 12L: 12D), Acomys russatus exhibits a tendency towards a nocturnal activity pattern, compared to the diurnal activity displayed by this species under natural conditions. Under the same photoperiod regime and at T_a = 28^oC, Apodemus mystacinus displays nocturnal activity, as observed under natural conditions. The maximal values of T_b were recorded in Acomys russatus at midnight (23:50 h), while the maximal values of VO₂ were recorded at the beginning of the dark period (18:20 h). In Apodemus mystacinus, the maximal values of T_b and VO₂ were recorded at 23:40 and 20:00 h, respectively. The ecophysiological significance of these results is discussed further.     

Differential effects of multiple short day lengths on body weights of gonadectomized siberian hamsters

Siberian hamsters (Phodopus sungorus) maintained under simulated natural photoperiods exhibit marked reductions in body weight as day lengths decrease in summer and fall. This experiment assessed whether the component of the seasonal body weight rhythm that is independent of gonadal hormones exhibits a graded dependence on decreasing day lengths or whether the entire program of weight loss is triggered by the crossing of a single critical day length in late summer. Male hamsters born into a photoperiod with 13 h light and 11 h dark (i.e., 13L : 11D) were castrated and transferred to simulated natural photoperiod for early April at 40 degrees N latitude. At the summer solstice (15L : 9D) some hamsters remained on that photoperiod whereas others experienced gradual decreases in day length. Three additional groups were moved to static photoperiods when day lengths had subsequently declined to 13L : 11D, 11L : 13D, or 9L : 15D, respectively. Day lengths decreasing to 13L : 11D were sufficient to suppress body weight but were less inhibitory than further decreases in day length. Hamsters identified as photononresponsive on the basis of daily activity rhythms increased body weight monotonically. These results establish that steroid-independent modulation of body weight depends on photoperiod in a graded fashion.     

Photic and Pineal Modulation of Food Anticipatory Circadian Activity Rhythms in Rodents

Restricted daily feeding schedules entrain circadian oscillators that generate food anticipatory activity (FAA) rhythms in nocturnal rodents. The location of food-entrainable oscillators (FEOs) necessary for FAA remains uncertain. The most common procedure for inducing circadian FAA is to limit food access to a few hours in the middle of the light period, when activity levels are normally low. Although light at night suppresses activity (negative masking) in nocturnal rodents, it does not prevent the expression of daytime FAA. Nonetheless, light could reduce the duration or magnitude of FAA. If so, then neural or genetic ablations designed to identify components of the food-entrainable circadian system could alter the expression of FAA by affecting behavioral responses to light. To assess the plausibility of light as a potential mediating variable in studies of FAA mechanisms, we quantified FAA in rats and mice alternately maintained in a standard full photoperiod (12h of light/day) and in a skeleton photoperiod (two 60 min light pulses simulating dawn and dusk). In both species, FAA was significantly and reversibly enhanced in the skeleton photoperiod compared to the full photoperiod. In a third experiment, FAA was found to be significantly attenuated in rats by pinealectomy, a procedure that has been reported to enhance some effects of light on behavioral circadian rhythms. These results indicate that procedures affecting behavioral responses to light can significantly alter the magnitude of food anticipatory rhythms in rodents.     

Circadian and photoperiodic time measurement in male Syrian hamsters following lesions of the melatonin-binding sites of the paraventricular thalamus.

Autoradiographic studies using [125I]iodomelatonin in several species, including the Syrian hamster, have revealed that the rostral region of the anterior paraventricular nucleus of the thalamus (aPVT) contains a very high density of binding sites for melatonin. In two studies, small or large bilateral electrolytic lesions of the aPVT were made in adult male hamsters maintained on long days (LD 16:8). The hamsters were then transferred to short days (LD 8:16) to test whether testicular regression could occur in response to a decrease in photoperiod. Serum prolactin concentrations were measured as a second photoperiodic response. All unoperated control hamsters showed the typical short-day photoperiodic response: A decrease in serum luteinizing hormone (LH) and prolactin concentrations and testicular regression all occurred within 6 weeks in short days, followed by the development of scotorefractoriness. Lesions of the aPVT did not significantly affect the rate or the degree of the short-day-induced decline in serum levels of LH or prolactin, nor the pattern of testicular regression and the subsequent expression of refractoriness. To enable us to determine whether the aPVT might be involved in the entrainment or the expression of circadian rhythms, locomotor activity was monitored continuously in lesioned and control groups in Experiment 2, prior to and following the switch to short days. The reduction in photoperiod (involving an 8-hr advance in the time of lights-off and an 8-hr extension of the dark phase) caused a decompression of the nocturnal activity bout of control animals, so that after 2 weeks in short days, activity onset had also advanced to regain its phase relationship to the timing of lights-off. A similar pattern of reentrainment was observed in lesioned animals, and no differences were observed between treatment groups in the rate of entrainment and decompression. In addition, both intact controls and animals bearing large bilateral lesions of the aPVT exhibited robust free-running circadian rhythms of locomotor activity when held under constant dim red light. In summary, the integrity of the aPVT is not necessary for the seasonal response of the reproductive axis and prolactin secretion to photoperiod, nor for photic entrainment of activity rhythms, in the Syrian hamster.     

Photoperiodic influences on ultradian rhythms of male siberian hamsters

Seasonal changes in mammalian physiology and behavior are proximately controlled by the annual variation in day length. Long summer and short winter day lengths markedly alter the amplitude of endogenous circadian rhythms and may affect ultradian oscillations, but the threshold photoperiods for inducing these changes are not known. We assessed the effects of short and intermediate day lengths and changes in reproductive physiology on circadian and ultradian rhythms of locomotor activity in Siberian hamsters. Males were maintained in a long photoperiod from birth (15 h light/day; 15 L) and transferred in adulthood to 1 of 7 experimental photoperiods ranging from 14 L to 9 L. Decreases in circadian rhythm (CR) robustness, mesor and amplitude were evident in photoperiods ≤14 L, as were delays in the timing of CR acrophase and expansion of nocturnal activity duration. Nocturnal ultradian rhythms (URs) were comparably prevalent in all day lengths, but 15 L markedly inhibited the expression of light-phase URs. The period (τ'), amplitude and complexity of URs increased in day lengths ≤13 L. Among hamsters that failed to undergo gonadal regression in short day lengths (nonresponders), τ' of the dark-phase UR was longer than in photoresponsive hamsters; in 13 L the incidence and amplitude of light-phase URs were greater in hamsters that did not undergo testicular regression. Day lengths as long as 14 L were sufficient to trigger changes in the waveform of CRs without affecting UR waveform. The transition from a long- to a short-day ultradian phenotype occurred for most UR components at day lengths of 12 L-13 L, thereby establishing different thresholds for CR and UR responses to day length. At the UR-threshold photoperiod of 13 L, differences in gonadal status were largely without effect on most UR parameters.     

Seasonal variations in circadian rhythms coincide with a phase of sensitivity to short photoperiods in the European hamster

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.     

Light-Dark Entrainment of Circadian Activity Rhythms of the Diurnal Indian Palm Squirrel (Funambulus pennanti)

A recent focus of chronobiological studies has been to establish diurnal models as alternatives to the more frequently used nocturnal rodents. In the present study, light-dark (LD) entrainment characteristics were examined in one diurnal species, the Indian palm squirrel (Funambulus pennanti). Palm squirrels showed strongly diurnal locomotor activity rhythms (? 88 percent) under light-dark (LD) cycles, with activity bimodally distributed during the L phase. In comparison to a dim LD cycle, exposure to a bright LD cycle caused a phase advance in the onset of activity, an increase in daily activity levels and an increase in the duration of activity. Percentage diurnality, however, did not vary between bright and dim LD cycles. Activity rhythms reentrained in significantly fewer days after an 8 hour phase delay of the LD cycle compared to an 8 hour phase advance. In both cases, the direction of reentrainment followed the direction of the LD shift. When exposed to single light pulses (1 hour) presented at the same time each day, 6/7 squirrels entrained. Under a skeletal photoperiod cycle (2 x 1 hour light pulses each day), 6/8 squirrels showed stable entrainment. The remaining squirrels exhibited rhythm splitting, with each component synchronising in an unstable manner with one of the light pulses. Under entrainment to single light pulses and to the skeletal photoperiod cycle, the phase angle of entrainment was negatively correlated with t. Finally, when exposed to a skeletal scotoperiod cycle (2 x 1-hour dark pulses each day), only 3/8 squirrels entrained, while the others free-ran. Two of the entrained squirrels showed spontaneous phase reversals during entrainment. As with other species, the activity rhythm of palm squirrels appears to be controlled by two separate self-sustaining oscillators. The strongly diurnal nature of palm squirrels make them a promising diurnal model for studies examining endogenous and exogenous influences on circadian functioning.     

The pineal and melatonin in the regulation of pituitary-thyroid axis

Studies of thyroid physiology in rats and hamsters support the view that the pineal gland has an anti-thyrotropic action. While chronic exposure of hamsters to short photoperiod, darkness, or blindness results in a depression of plasma thyroxin and plasma TSH, removal of the pineal gland, which synthesizes melatonin, prevents these effects. Melatonin administration, in the form of daily injections given late in the photoperiod, also results in inhibition of plasma thyroxin and plasma TSH. These anti-thyrotropic effects are similar to the anti-gonadotropic effects of melatonin. The results of a variety of experiments are consistent with the view that melatonin acts on a neuroendocrine control mechanism influencing synthesis or release of hypothalamic thyrotropin releasing hormone (TRH).