Expression of circadian rhythmicity in Djungarian hamsters under constant light: Effects of light intensity and the circadian system's state

Summary Djungarian hamsters (Phodopus sungorus), were exposed to constant light with increasing intensities (20, 60, 350 lux), and wheel running activity was recorded. With increasing light intensity the percentage of hamsters showing a split in their daily activity pattern increased and the free running period was lengthened for both the unsplit and the split state. The fact that the free running period of both states depended on the light intensity together with the observation that the highest incidence of acircadian activity occurred under 350 lux, provoked the idea that the emergence of splitting or acircadian rhythmicity is a direct consequence of the light induced lengthening of the free running period. However, analysis of the data failed to support the idea that emergence of a split or acircadian activity is a threshold phenomenon with respect to the free running period.Due to differences in circadian function some Djungarian hamsters do not exhibit photoinduction following short day exposure. In these individuals splitting also occurred but required exposure to a higher light intensity than in photo-responsive hamsters. This observation is in accordance with the idea that the two phenotypes differ in the interaction of the two component oscillators underlying circadian rhythmicity.Abbreviations LD long day photoperiod - LL constant light - SD short day photoperiod - free running period     

Non-parametric photic entrainment of Djungarian hamsters with different rhythmic phenotypes

To investigate the role of non-parametric light effects in entrainment, Djungarian hamsters of two different circadian phenotypes were exposed to skeleton photoperiods, or to light pulses at different circadian times, to compile phase response curves (PRCs). Wild-type (WT) hamsters show daily rhythms of locomotor activity in accord with the ambient light/dark conditions, with activity onset and offset strongly coupled to light-off and light-on, respectively. Hamsters of the delayed activity onset (DAO) phenotype, in contrast, progressively delay their activity onset, whereas activity offset remains coupled to light-on. The present study was performed to better understand the underlying mechanisms of this phenomenon. Hamsters of DAO and WT phenotypes were kept first under standard housing conditions with a 14:10 h light–dark cycle, and then exposed to skeleton photoperiods (one or two 15-min light pulses of 100 lx at the times of the former light–dark and/or dark–light transitions). In a second experiment, hamsters of both phenotypes were transferred to constant darkness and allowed to free-run until the lengths of the active (α) and resting (ρ) periods were equal (α:ρ = 1). At this point, animals were then exposed to light pulses (100 lx, 15 min) at different circadian times (CTs). Phase and period changes were estimated separately for activity onset and offset. When exposed to skeleton-photoperiods with one or two light pulses, the daily activity patterns of DAO and WT hamsters were similar to those obtained under conditions of a complete 14:10 h light–dark cycle. However, in the case of giving only one light pulse at the time of the former light–dark transition, animals temporarily free-ran until activity offset coincided with the light pulse. These results show that photic entrainment of the circadian activity rhythm is attained primarily via non-parametric mechanisms, with the “morning” light pulse being the essential cue. In the second experiment, typical photic PRCs were obtained with phase delays in the first half of the subjective night, phase advances in the second half, and a dead zone during the subjective day. ANOVA indicated no significant differences between WT and DAO animals despite a significantly longer free-running period (tau) in DAO hamsters. Considering the phase shifts induced around CT0 and the different period lengths, it was possible to model the entrainment patterns of both phenotypes. It was shown that light-induced phase shifts of activity offset were sufficient to compensate for the long tau in WT and DAO hamsters, thus enabling a stable entrainment of their activity offsets to be achieved. With respect to activity onsets, phase shifts were sufficient only in WT animals; in DAO hamsters, activity onset showed increasing delays. The results of the present paper clearly demonstrate that, under laboratory conditions, the non-parametric component of light and dark leads to circadian entrainment in Djungarian hamsters. However, a stable entrainment of activity onset can be achieved only if the free-running period does not exceed a certain value. With longer tau values, hamsters reveal a DAO phenotype. Under field conditions, therefore, non-photic cues/zeitgebers must obviously be involved to enable a proper circadian entrainment.     

Evidence for independence of circadian characters and extent of photoresponsiveness in the Djungarian hamster, Phodopus sungorus.

Djungarian hamsters (Phodopus sungorus) exposed to a short-day photoperiod generally respond with a syndrome of physiological and behavioral changes, such as body weight loss and molt to a white pelage. The extent of the short-day-induced responses differs among individuals. Furthermore, some hamsters show no photoresponse. In this study, we sought to determine whether variation in the photoresponse would be associated with circadian function: whether phase angle or free-running period (tau) would differ between responsive and nonresponsive hamsters; and whether changes in these circadian characters would correlate with the extent of weight loss and molt (and the timing of molt onset) in photoresponsive hamsters. Adult hamsters were kept in a short-day photoperiod (9 hr light, 15 hr dark) for 14 weeks, during which time body weight and molt were measured biweekly. Hamsters were then transferred to cages equipped with running wheels; we measured the phase angle of activity onset under a short-day photoperiod and tau in constant dark. Hamsters exhibiting a short-day-induced molt had a significantly shorter tau and a less negative phase angle than nonmolting animals. Hamsters that exhibited weight loss also had a significantly less negative phase angle, but no difference in tau. No significant Pearson's or Spearman's correlation coefficients were found between extent (or timing) of the photoresponse and the circadian characters in responsive hamsters. Although these results indicate that threshold for photoresponsiveness is related to circadian function, the extent (and timing) of the photoresponse may not be.     

Photobiology of the Gonyaulax circadian system. I. Different phase response curves for red and blue light

Phase responses to red and blue light pulses were measured at different times during the circadian cycle (phase response curves, PRC) in the marine unicellular dinoflagellate Gonyaulaxpolyedra Stein. Pulses were given during a 24-h period of darkness; thereafter, cultures were released into constant dim red light for the assessment of phase and period. The results confirmed earlier findings that the Gonyaulax circadian system receives light signals via two distinct input pathways. During the subjective day and for the first 3 h of the subjective night, red and blue light pulses led to identical phase responses. For the rest of the circadian cycle, however, phase responses to pulses of either red or blue light differed drastically both in their amplitude and direction (advances or delays). Thus, the Gonyaulax light PRC is generated by two distinct light responses. One of these represents responses via a light input that is responsive both to red and blue light mainly producing small delays. The other represents responses of a primarily blue-sensitive input system leading to large advances restricted to the subjective night. Via feed-back, the blue-sensitive light input appears to be under the control of the circadian system. Received: 27 November 1996/Accepted: 30 January 1997     

Circadian phase response curves for dark pulses in the hamster

Summary Hamsters maintained under constant illumination were exposed to 2- or 6-h pulses of darkness at various phases of their circadian activity rhythms. When presented around the time of activity onset, the pulses resulted in phase advances, and when presented toward the end of daily activity, they resulted in phase delays. Since others have shown that light pulses presented at the same phases in constant darkness cause phase shifts in the opposite directions, these results indicate that phase response curves for light and dark pulses are mirror images.Dark pulses also caused phase-dependent changes, both transient and long-lasting, in the period of the free-running rhythms, and a few pulses were immediately followed by splitting of the activity rhythms into two components. Such effects may reflect a differential responsiveness of two coupled oscillators to dark pulses.Abbreviations CT circadian time - DD constant dark - LD lightdark - LL constant light - PRC phase response curve - SD subjective day - SN subjective night - period of a circadian rhythm Supported by grants from the NSERC of Canada to B. Rusak and to G.V. Goddard. We are grateful to Dr. Goddard for his support and encouragement     

Circadian and photoperiodic effects of brief light pulses in male Djungarian hamsters

The effects of brief light pulses (1-60 min in duration) on the circadian rhythm of locomotor activity and/or the neuroendocrine-gonadal axis was investigated in male Djungarian hamsters. Exposure of hamsters free-running in constant darkness to a single 1-h pulse of light induced phase-dependent phase shifts in the rhythm of locomotor activity. The general shape of the "phase-response curve" was similar to that observed in other animals; phase-delays and phase-advances were induced by light pulses delivered in the early and late subjective night, respectively, while light pulses during the subjective day induced little or no phase-shift in the activity rhythm. Animals exposed for 7 days to 1-min of light during the night in animals otherwise exposed to 6L:18D resulted in increased levels of serum FSH and testicular weight. Daily exposure to two 1-h or two 10-min pulses of light (but not two 1-min pulses) for 10 days resulted in stable entrainment of the activity rhythm as well as testicular weight gains and serum FSH increases. When two 10-min pulses of light were presented 8 and 16 h apart, some animals showed a short-day entrainment pattern (i.e., locomotor activity confined to the long period of darkness) while other animals showed a long-day entrainment pattern (i.e., locomotor activity confined to the short period of darkness). Importantly, the stimulatory effects of light on neuroendocrine-gonadal activity were clearly dependent on the phase-relationship between the light pulses and the circadian rhythm of locomotor activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Short-Day Response in Djungarian Hamsters of Different Circadian Phenotypes

In Djungarian hamsters (Phodopus sungorus) bred at the authors' institute, a certain number of animals show activity patterns incompatible with proper entrainment of their endogenous circadian pacemaker to the environmental light-dark (LD) cycle. Even though the activity-offset in these animals is stably coupled to “light-on,” activity-onset is increasingly delayed, leading to a compression of the activity time (α). If α falls below a critical value, the circadian rhythm in these so called delayed activity-onset (DAO) hamsters starts to free-run and finally breaks down. Animals then show an arrhythmic activity pattern (AR hamsters). Previous studies revealed the mechanisms of photic entrainment have deteriorated (DAO) or the suprachiasmatic nucleus (SCN) does not generate a rhythmic signal (AR). The aim of the present study was to investigate the consequences that these deteriorations have upon photoperiodic time measurement. Animals were bred and kept under standardized housing conditions with food and water ad libitum and a 14L/10D (long day, LD) regimen. Locomotor activity was recorded continuously using passive infrared motion detectors. Body mass, testes size, and fur coloration were measured weekly or biweekly to further quantify the photoperiodic reaction. In a first experiment, adult male wild-type (WT), DAO, and AR hamsters were transferred initially to a 16L/8D cycle. After 3–4 wks, the light period was shortened symmetrically by 8?h. After 14 wks, none of the DAO and AR hamsters, and only 1 of 8 WT hamsters showed short-day (SD) traits. Therefore, in a second experiment, hamsters were transferred to SD conditions (8L/16D cycle) for 8 wks directly from standard LD conditions. In 6 of 7 WT hamsters, activity time expanded, body mass and testes size decreased, and fur coloration changed from summer to winter pelage. In contrast, none of the DAO and AR hamsters displayed an SD response. In a third experiment, DAO and AR hamsters were kept in constant darkness (DD) for 8 and 14 wks. After 8 wks, DAO hamsters showed a similar photoperiodic reaction to WT hamsters that had been kept for 8 wks under SD conditions. However, the level of adaptation was still less compared to WT hamsters, but this difference was not apparent after 14 wks. In contrast, AR animals did not display any photoperiodic reaction, even after 14 wks in DD. Type VI phase response curves (PRCs) were constructed to better understand the mechanism behind the SD response. In WT hamsters, the photosensitive phase, where light pulses induce phase shifts, was lengthened in SD condition. In DAO hamsters, in contrast, the PRCs were similar under LD and SD conditions with a compressed photosensitive phase corresponding to α. Also, “light-on” induced only weak phase advances of activity-onset, insufficient to compensate for the long endogenous period. The results show that physiological mechanisms necessary for seasonal adaptation are working in DAO hamsters and that it is the inadequate interaction of the LD cycle with the SCN that prevents the photoperiodic reaction. AR hamsters, on the other hand, are incapable of measuring photoperiodic time due to a complete disruption of circadian rhythmicity.     

Light-induced c-Fos expression in the SCN and behavioural phase shifts of Djungarian hamsters with a delayed activity onset

C-Fos expression in the suprachiasmatic nucleus (SCN) and phase shifts of the activity rhythm following photic stimulation were investigated in Djungarian hamsters (Phodopus sungorus) of two different circadian phenotypes. Wild-type (WT) hamsters display robust daily patterns of locomotor activity according to the light/dark conditions. Hamsters of the DAO (delayed activity onset) phenotype, however, progressively delay the activity onset, whereas activity offset remains coupled to “light-on”. Although the exact reason for the delayed activity onset is not yet clarified, it is connected with a disturbed interaction between the light/dark cycle and the circadian clock. The aim was to test the link between photoreception and the behavioral output of the circadian system in hamsters of both phenotypes, to get further insight in the underlying mechanism of the DAO phenomenon. Animals were exposed to short light pulses at different times during the dark period to analyze phase shifts of the activity rhythm and expression of Fos protein in the SCN. The results indicate that the photosensitive phase in DAO hamsters is shifted like the activity onset. Also, phase shifts were significantly smaller in DAO hamsters. At the same time, levels of Fos expression did not differ between phenotypes regarding the circadian phase. The results provide evidence that the shifted photosensitivity of the circadian system in DAO hamsters does not differ from that of WT animals, and lead us to conclude that processes within the SCN that enable light information to reset the circadian pacemaker might offer an explanation for the DAO phenomenon.     

A short red light pulse during dark phase of LD-cycle perturbs the hamster's circadian clock

In this study we investigated the influence of red light, which naturally occurs during dawn and dusk, on locomotor activity and body temperature rhythms of Djungarian hamsters (Phodopus sungarus). A single weak red light pulse given 2 h before regular lights on had acute as well as long-term effects persisting for several days following exposure. The hamsters immediately stopped their locomotor activity, accompanied by a drop in body temperature. In the following undisturbed nights (LD 168) the nocturnal activity stopped earlier than usual. This lasting effect of the light pulse was more pronounced than the acute effect. The activity phase compressed gradually during 3 to 5 days after the light pulse was administered while time of activity onset was almost unaffected. It took 6 to 11 days for complete recovery of the original activity phase. The maximal activity compression and the recovery period depended on the duration of the single red light pulse and its intensity. Red light pulses of 15 min duration were about twice effective as 1 min pulses; and the effect of a red light pulse of 130 mW/m² was about 1.5 times stronger than a 30 mW/m² red light pulse. The maximal value of activity phase compression reached in this experiment was 2.5+0.2 h with a recovery period of 11.1±0.3 days following a given red light pulse of 90 mW/m² and 15 min. The morning oscillator seems to be persistently affected. This indicates a very high photosensitivity of the Djungarian hamster's circadian system to red light.Abbreviations T _b body temperature - DD constant darkness - LD light:dark cycle - LL constant light - duration of activity phase - CT circadian time - PRC phase response curve - SCN suprachiasmatic nuclei     

c-Fos expression in the brains of behaviorally "split" hamsters in constant light: calling attention to a dorsolateral region of the suprachiasmatic nucleus and the medial division of the lateral habenula

"Splitting" of circadian activity rhythms in Syrian hamsters maintained in constant light appears to be the consequence of a reorganized SCN, with left and right halves oscillating in antiphase; in split hamsters, high mRNA levels characteristic of day and night are simultaneously expressed on opposite sides of the paired SCN. To visualize the splitting phenomenon at a cellular level, immunohistochemical c-Fos protein expression in the SCN and brains of split hamsters was analyzed. One side of the split SCN exhibited relatively high c-Fos levels, in a pattern resembling that seen in normal, unsplit hamsters during subjective day in constant darkness; the opposite side was labeled only within a central-dorsolateral area of the caudal SCN, in a region that likely coincides with a photo-responsive, glutamate receptor antagonist-insensitive, pERK-expressing cluster of cells previously identified by other laboratories. Outside the SCN, visual inspection revealed an obvious left-right asymmetry of c-Fos expression in the medial preoptic nucleus and subparaventricular zone of split hamsters killed during the inactive phase and in the medial division of the lateral habenula during the active phase (when the hamsters were running in their wheels). Roles for the dorsolateral SCN and the mediolateral habenula in circadian timekeeping are not yet understood.     

Running activity mediates the phase-advancing effects of dark pulses on hamster circadian rhythms

Summary Pulses of darkness can phase-shift the circadian activity rhythms of hamsters,Mesocricetus auratus, kept in constant light. Dark pulses under these conditions alter photic input to the circadian system, but they also commonly trigger wheel-running activity. This paper investigates the contribution of running activity to the phase-shifting effects of dark pulses. A first experiment showed that running activity by itself can phaseshift rhythms in constant light. Hamsters were induced to run by being confined to a novel wheel for 3–5 h. When this was done at circadian times (CT) 0, 6, and 9, the mean steady-state phase-shifts were 0.6 h, 3.5 h, and 2.3 h, respectively. The latter two values are at least as large as those previously obtained with dark pulses of similar durations and circadian phases. A second experiment showed that restricting the activity of hamsters during 3-h dark pulses at CT 9 reduces the amplitude of the phase-shifts. Unrestrained animals phase-advanced by 1.1 h, but this shift was halved in animals whose wheel was locked, and completely abolished in animals confined to nest boxes during the dark pulse. Activity restriction in itself (without dark pulses) had only minimal phase-delaying effects on free-running rhythms when given between ca. CT 10 and CT 13. These results support the idea that, in hamsters at least, dark pulses affect the circadian system mostly by altering behavioural states rather than by altering photic input to the internal clock.Abbreviations CT circadian time - DD constant darkness - LD light-dark - LL constant light - PRC phase response curve - period of rhythm     

Lesions of the thalamic intergeniculate leaflet alter hamster circadian rhythms

We have investigated the effects of destruction of the geniculo-hypothalamic tract (GHT) on the circadian system of golden hamsters. In the first experiment, intact hamsters were housed in constant darkness, and phase shifts in running-wheel activity rhythms were assessed following 15-min light pulses administered at circadian time (CT) 12 (defined as the beginning of activity), CT 14, CT 18, and CT 20. Responses to light pulses at the same CTs were then reassessed after GHT lesions. Hamsters with complete lesions showed decreases in phase advances caused by light pulses at CT 18 and CT 20. Phase delays elicited by light at CT 12 and CT 14 were not altered. In a second study, intact and GHT-ablated hamsters housed in constant light received 6-hr dark pulses at various CTs. Hamsters with complete GHT ablation showed smaller advances than controls to dark pulses centered on CT 8-10. After 110 days in constant light, 7 of 10 intact hamsters showed splitting of their activity rhythms into two components, while only 1 of the 8 similarly treated ablated hamsters displayed dissociated activity components. Ablated hamsters had significantly shorter free-running periods during the first 35 days of exposure to constant light than did the intact hamsters. These results demonstrate that destruction of the GHT in the hamster alters phase shifting in response to periods of light or dark, and they indicate a role for the GHT in mediating several photic effects on the circadian system.     

Exotic photoperiods induce and entrain split circadian activity rhythms in hamsters

The split circadian activity rhythm that emerges in hamsters after prolonged exposure to constant light has been a theoretical cornerstone of a multioscillator view of the mammalian circadian pacemaker. The present study demonstrates a novel method for splitting hamster circadian rhythms and entraining them to exotic light:dark cycles. Male Syrian hamsters previously maintained on a 14-h day and 10-h night were exposed to a second 5-h dark phase in the afternoon. The 10-h night was progressively shortened until animals experienced two 5-h dark phases beginning 10 h apart. Most hamsters responded by splitting their activity rhythms into two components associated with the afternoon and nighttime dark phases, respectively. Each activity component was entrained to this light:dark:light:dark cycle. Transfer of split hamsters to constant darkness resulted in rapid joining of the two activity components with the afternoon component associated with onset of the fused rhythm. In constant light, the nighttime component corresponded to activity onset of the fused rhythm, but splitting emerged again at an interval characteristic for this species. The results place constraints on multi-oscillator models of circadian rhythms and offer opportunities to characterize the properties of constituent circadian oscillators and their interactions.     

Entrainment of 2 subjective nights by daily light:dark:light:dark cycles in 3 rodent species

Recent work with exotic 24-h light:dark:light:dark (LDLD) cycles indicates surprising flexibility in the entrainment patterns of Syrian hamsters. Following exposure to an LDLD cycle, hamsters may adopt a form of rhythm splitting in which markers of subjective night (e.g., activity, melatonin) are expressed in each of the twice daily scotophases. This pattern contrasts markedly with that of conventionally entrained hamsters in which markers of subjective night are expressed once daily in only 1 of the 2 dark periods. The "split" entrainment pattern was examined further here in Syrian and Siberian hamsters and in mice exposed to LDLD 7:5:7:5, a condition that reliably induces split activity rhythms in all 3 species. The phase angle of entrainment and activity duration were generally similar comparing the 2 daily activity bouts in each species. The stability of this split entrainment state was assessed by deletions of photophases on individual days, by exposure to skeleton photoperiods, and by transfer to constant darkness. As in Syrian hamsters, the one-time substitution of darkness for one 7-h photophase did not grossly alter activity patterns of Siberian hamsters but acutely disrupted the split rhythms of mice. Skeleton light pulses of progressively shorter duration did not significantly alter split entrainment patterns of either Syrian or Siberian hamsters. Both species continued to exhibit stable entrainment with activity expressed in alternate scotophases of an LD 1:5 cycle presented 4 times daily. In contrast, the split activity rhythms of mice were not maintained under skeleton pulses. In constant darkness, rhythms of Siberian hamsters remained distinctly split for a minimum of 2 cycles. Split entrainment to these novel LDLD and 4-pulse skeleton lighting regimes demonstrates a marked degree of plasticity common to the circadian systems of several rodent species and identifies novel entrainment patterns that may be reliably elicited with simple environmental manipulations. Inter- and intraspecific differences in the stability of split activity rhythms likely reflect differences in coupling interactions between the component circadian oscillators, which, adopting separate phase relations to these novel LD cycles, yield a split entrainment pattern.     

Effects of light intensity and restraint on dark-pulse-induced circadian phase shifting during subjective night in Syrian hamsters

Dark pulses presented on a background of constant light (LL) result in phase advances during midsubjective day and early subjective night, and phase delays during late subjective night, as shown in the dark-pulse phase response curve. In hamsters, the phase-shifting effects of dark pulses are thought to be mediated by increased activity, as previous studies have shown that restraining animals during dark pulses blocks the phase shifts observed in midsubjective day and late subjective night. This study focuses on dark-pulse-induced phase shifting during early subjective night, examining the influence of both LL intensity and restraint on the magnitude of these phase shifts. Syrian hamsters were maintained in LL of four different illumination levels (1, 10, 100, or 600 lux) and periodically presented with 6-h pulses (dark pulse alone, restraint alone, or dark pulse plus restraint) beginning at circadian time 11. Phase advances were observed in response to dark pulses alone, and the magnitude of these shifts was dependent on background illumination, with significantly larger advances seen under higher intensities. No relationship was found between the amount of activity displayed during dark pulses and phase shift magnitude. Six-hour periods of restraint resulted in phase delays, the magnitude of which was also dependent on background illumination. Restraining hamsters during dark pulses reduced the magnitude of phase advances, but the extent of this reduction could be predicted from the additive effects of the dark-pulse-alone and restraint-alone conditions. These results indicate that the phase-shifting effects of dark pulses during early subjective night are not mediated by behavioral activation and may instead reflect a mirror image of the phase-delaying effects of light pulses at this phase.     

Effects of Clomipramine Administration on Syrian Hamster Circadian System and Behavior

The aim of the present work is to discuss the available data on neonatal and adult antidepressant treatment in relation to animal models of depression and serotonergic modulation of the circadian system, with a particular emphasis on our own published and unpublished work on the effects of clomipramine (a serotonin reuptake inhibitor) on the Syrian hamster circadian behavior. Neonatal clomipramine treatment (15 mg/kg from postnatal days 8 to 21) significantly augmented the amplitude of the wheel running rhythm, as well as delayed its acrophase and increased the time to reentrain after a 6-h phase advance of the light-dark cycle. Neonatally clomipramine-treated hamsters had a shorter circadian period than saline-treated animals under constant light - but not under constant dark- conditions, exhibited decreased phase advances after light pulses applied at late subjective night and greater phase advances after i.p. administration of the 5-HT_1A-receptor agonist 8-OH-DPA at midday. These animals also exhibited more locomotor activity than controls, but did not display the typical circadian variation in anxiety-related behavior, as measured in a plus-maze paradigm. They also showed an increased 5-HIAA/5-HT ratio in hypothalamus and midbrain raphe, while 5-HT content was decreased in frontal cortex and anterior hypothalamic areas. Since drugs linked to the serotonergic system are able to modify the circadian system, we decided to test whether acute and chronic clomipramine administration in adulthood was able to change: a) the phase of free running activity rhythms; (b) light-induced phase shifts, and (c) hypothalamic 5-HT turnover. Acute clomipramine injection had a phase-dependent effect on the free running activity rhythm, with phase advances at CT 0-8 being significantly higher than at CT 8-16. Pretreatment with clomipramine inhibited phase advances in response to light pulses when applied at CT 19 while phase delays at CT 14 remained unaffected. This acute treatment also decreased 5-HT turnover in the SCN at both CTs. In contrast, chronic clomipramine administration potentiated light-induced phase advances, without changes in period, amplitude or central 5-HT turnover. Taken together, these data support the view that clomipramine, as other antidepressant drugs, can affect the expression of the circadian rhythmicity in Syrian hamsters, possibly through serotonergic mechanisms in the case of acute treatments, and more complex behavioral interaction in the case of neonatal and chronic treatments.     

Djungarian hamsters: a species with a labile circadian pacemaker? Arrhythmicity under a light-dark cycle induced by short light pulses

In most cases, phase-shifting effects of light pulses are studied in animals kept in constant darkness (DD) or in animals released into DD following the stimulus. In this study, the authors exposed Djungarian hamsters (Phodopus sungorus) to short light pulses during the dark phase of a 16:8 light-dark (LD) cycle and thus obtained a type VI phase response curve. Light pulses early in the night caused phase delays of the activity onset as well as phase advances of the activity offset, whereas light pulses later in the night resulted in phase advances of the activity offset only. A combination of two 15-min light pulses-the first one given late in the scotophase and the second given early in the dark phase of the following night-led to a strong compression of the activity phase alpha. In 75% of all animals, daily rhythms were no longer visible after complete alpha compression, and long-term arrhythmicity (up to 145 days) persisted despite continued exposure to an LD cycle. Because three independent output rhythms of the clock (i.e., activity, body temperature, and melatonin rhythms) were equally affected, the authors conclude that overt arrhythmicity was due not merely to disrupted output pathways but to an altered state of the central pacemaker. The authors suggest a qualitative two-oscillator model to explain this phenomenon. Their hypothesis assumes that, due to loose coupling, the pacemaker of Djungarian hamsters can be driven to a state of zero phase difference between the two oscillators, with zero amplitude of their outputs.     

Daily novel wheel running reorganizes and splits hamster circadian activity rhythms.

The phenomenon of splitting of locomotor activity rhythms in constant light has implied that the mammalian circadian pacemaker is composed of multiple interacting circadian oscillators. Exposure of male Syrian hamsters to novel running wheels also induces splitting in some reports, although novel wheel running (NWR) is better known for its effects on altering circadian phase and the length of the free-running period. In three experiments, the authors confirm and extend earlier reports of split rhythms induced by NWR. Male Syrian hamsters, entrained to LD 14:10, were transferred for 6 to 11 consecutive days to darkened novel Wahmann wheels at ZT 4 and were returned to their home cages at ZT 9. All hamsters ran robustly in the novel wheels. NWR caused a marked reorganization of home cage wheel-running behavior: Activity onsets delayed progressively with each additional day of NWR. After 11 days, activity onset in the nighttime scotophase was delayed by 7 h and disappeared completely in 2 hamsters (Experiment 1). After 6 to 7 days of NWR (Experiment 2), activity onset delayed by 5 h. Transfer of hamsters to constant darkness (DD) after 7 days of NWR revealed clearly split activity rhythms: The delayed nighttime activity bout was clearly identifiable and characterized by a short duration. A second bout associated with the former time of NWR was equally distinct and exhibited a similarly short duration. These components rejoined after 3 to 5 days in DD accomplished via delays and advances of the nighttime and afternoon components, respectively. The final experiment established that rejoining of activity components could be prevented by perpetuating the light-dark:light-dark cycle used to induce split rhythms. The data suggest that NWR causes selective phase shifting of some circadian oscillators and that component oscillators interact strongly in constant darkness.     

Age-related changes in circadian responses to dark pulses

Aging involves many alterations in circadian rhythms, including a loss of sensitivity to both photic and nonphotic time signals. This study investigated the sensitivity of young and old hamsters to the phase advancing effect of a 6-h dark pulse on the locomotor activity rhythm. Each hamster was tested four times during a period of approximately 9 mo; periods of exposure to a 14-h photoperiod were alternated with the periods of exposure to constant light (20-80 lx), during which the dark pulses were administered. There was no significant difference in the phase shifts exhibited by the young (4-10 mo) and old hamsters (19-25 mo) or in the amount of wheel running activity displayed during each dark pulse. However, young hamsters had a significantly greater propensity to exhibit split rhythms immediately after the dark pulses. These results suggest that, although aging does not reduce the sensitivity of the circadian pacemaker to this nonphotic signal, it alters one property of the pacemaker, i.e., the flexibility of the coupling of its component oscillators.     

Large phase-shifts of circadian rhythms caused by induced running in a re-entrainment paradigm: The role of pulse duration and light

Summary Bouts of induced wheel-running, 3 h long, accelerate the rate of re-entrainment of hamsters' activity rhythms to light-dark (LD) cycles that have been phase-advanced by 8 h (Mrosovsky and Salmon 1987). The bouts of running are given early in the first night of the new LD cycle, and by the second night the phase advance in activity onset already averages 7 h. Such large shifts contrast with the mean phase advance of <1 h at the peak of the phase response curve when hamsters in constant darkness (DD) experience 2-h pulses of induced activity (Reebs and Mrosovsky 1989). The present paper investigates pulse duration and light as possible causes for the discrepancy in shift amplitude between these two studies. In a first experiment, pulses of induced wheel-running 1 h, 3 h, or 5 h long were given at circadian times (CT) 6 and 22-2 to hamsters free-running in DD. Pulses given at CT 6 caused phase-advances of up to 2.8 h, whereas pulses at CT 22-2 resulted in delays of up to 1.0 h. Shifts after 3-h and 5-h pulses did not differ, but were larger than after 1-h pulses, and larger than after the 2-h pulses given in DD by Reebs and Mrosovsky (1989). Thus 3 h appears to be the minimum pulse duration necessary to obtain maximum phase-shifting effects. In a second experiment, the re-entrainment design of Mrosovsky and Salmon (1987) was repeated with the light portion of the shifted LD cycle eliminated. Hamsters exercised for 3 h phase-advanced 2.9 h on average (excluding 2 animals who ran poorly). When the same hamsters were exposed 7 days later to a 14-h light pulse starting 5 h after their activity onset, they advanced by an average of 3.3 h. Adding the average values for activity-induced shifts and light-induced shifts gives a total of about 6 h. Possible synergism between the effects of induced activity and those of light may account for the remaining small difference between this total and the 7-h advances previously reported.Abbreviations CT circadian time - DD constant darkness - LD light-dark - PRC phase response curve - free-running period of rhythm