Suppression of circadian rhythms of pineal and testicular hormones following lithium treatment in normal and reversed light-dark cycles, constant light and constant dark in rats

The aim of the current investigation was to study the effect of lithium on circadian rhythms of pineal - testicular hormones by quantitations of pineal and serum serotonin, N-acetylserotonin and melatonin, and serum testosterone at four time points (06.00, 12.00, 18.00 and 24.00) of a 24-hr period under normal photoperiod (L:D), reversed photoperiod (D:L), constant light (L:L) and constant dark phase (D:D) in rats. Circadian rhythms were observed in pineal hormones in all the combinations of photoperiodic regimens, except in constant light, and in testosterone levels in all the photoperiodic combinations. Pineal and serum N-acetylserotonin and melatonin levels were higher than serotonin at night (24.00 hr), in natural L:D cycle, in reversed L:D cycle or similar to normal L:D cycle in constant dark phase, without any change in constant light. In contrast, testosterone level was higher in light phase (12.00 hr through 18.00 hr) than in the dark phase (24.00 hr through 06.00 hr) in normal L:D cycle, in reversed L:D cycle, similar to normal L:D cycle in constant dark (D:D), and reversed to that of the normal L:D cycle in constant light (L:L). Lithium treatment (2 mEq/kg body weight daily for 15 days) suppressed the magnitude of circadian rhythms of pineal and serum serotonin, N-acetylserotonin and melatonin, and testosterone levels by decreasing their levels at four time points of a 24-hr period in natural L:D or reversed D:L cycle and in constant dark (D:D). Pineal indoleamine levels were reduced after lithium treatment even in constant light (L:L). Moreover, lithium abolished the melatonin rhythms in rats exposed to normal (L:D) and reversed L:D (D:L) cycles, and sustained the rhythms in constant dark. But testosterone rhythm was abolished after lithium treatment in normal (L:D)/reversed L:D (D:L) cycle or even in constant light/dark. The findings indicate that the circadian rhythm exists in pineal hormones in alternate light - dark cycle (L:D/D:L) and in constant dark (D:D), but was absent in constant light phase (L:L) in rats. Lithium not only suppresses the circadian rhythms of pineal hormones, but abolishes the pineal melatonin rhythm only in alternate light - dark cycles, but sustains it in constant dark. The testosterone rhythm is abolished after lithium treatment in alternate light - dark cycle and constant light/dark. It is suggested that (a) normal circadian rhythms of pineal hormones are regulated by pulse dark phase in normal rats, (b) lithium abolishes pineal hormonal rhythm only in pulse light but sustains it in constant dark phase, and (c) circadian testosterone rhythm occurs in both pulse light or pulse dark phase in normal rats, and lithium abolishes the rhythm in all the combinations of the photoperiod. The differential responses of circadian rhythms of pineal and testicular hormones to pulse light or pulse dark in normal and lithium recipients are discussed.     

The experience in the water maze task can affect the circadian rhythm of locomotor activity

Wistar rats maintained in cages with running wheels and submitted to a skeleton photoperiod or to a light - dark cycle were tested in the Morris water maze. Half of the animals were exposed to the task during their active phase while the other half was exposed during their inactive phase. The effect of the experience in the water maze, a strong arousing event, on the rhythm of wheel-running activity was evaluated. In the first experiment, a group of animals submitted to a skeleton photoperiod was trained every day in the reference memory version of the task. The novel experience in the water maze had a strong phase-dependent masking effect: it produced an intense post-training bout of activity in the animals tested during their inactive phase. Another experiment was run using single working memory sessions in the water maze and with animals submitted to a light - dark cycle. The circadian rhythm of locomotor activity was evaluated on undisturbed days and compared with testing days. The experience in the water maze produced a significant increase in variability of activity onset during both circadian phases. Taken together, the data suggest that there is a modulating effect of the arousing experience in the pool on the overt circadian rhythm of locomotor activity.     

The effects of pinealectomy and blinding on the circadian locomotor activity rhythm in the Japanese newt,Cynops pyrrhogaster

We examined the effects of pinealectomy and blinding (bilateral ocular enucleation) on the circadian locomotor activity rhythm in the Japanese newt, Cynops pyrrhogaster. The pinealectomized newts were entrained to a light-dark cycle of 12 h light and 12 h darkness. After transfer to constant darkness they showed residual rhythmicity for at least several days which was gradually disrupted in prolonged constant darkness. Blinded newts were also entrained to a 12 h light/12 h dark cycle. In subsequent constant darkness they showed free-running rhythms of locomotor activity. However, the freerunning periods noticeably increased compared with those observed in the previous period of constant darkness before blinding. In blinded newts entrained to the light/dark cycle the activity rhythms were gradually disrupted after pinealectomy even in the presence of the light/dark cycle. These results suggest that both the pineal and the eyes are involved in the newt's circadian system, and also suggest that the pineal of the newt acts as an extraretinal photoreceptor which mediates the entrainment of the locomotor activity rhythm.Abbreviations circadian period - DD constant darkness - LD cycle, light-dark cycle - LD 12:12 light-dark cycle of 12 h light and 12 h darkness     

Circadian rhythms of locomotor activity and temperature selection in sleepy lizards, <Emphasis Type="Italic">Tiliqua rugosa</Emphasis>

This study examined whether the daily rhythms of locomotor activity and behavioural thermoregulation that have previously been observed in Australian sleepy lizards (Tiliqua rugosa) under field conditions are true circadian rhythms that persist in constant darkness (DD) and whether these rhythms show similar characteristics. Lizards held on laboratory thermal gradients in the Australian spring under the prevailing 12-hour light : dark (LD) cycle for 14 days displayed robust daily rhythms of behavioural thermoregulation and locomotor activity. In the 13-day period of DD that followed LD, most lizards exhibited free-running circadian rhythms of locomotor activity and behavioural thermoregulation. The predominant activity pattern displayed in LD was unimodal and this was retained in DD. While mean levels of skin temperature and locomotor activity were found to decrease from LD to DD, activity duration remained unchanged. The present results demonstrate for the first time that this species’ daily rhythm of locomotor activity is an endogenous circadian rhythm. Our results also demonstrate a close correlation between the circadian activity and thermoregulatory rhythms in this species indicating that the two rhythms are controlled by the same master oscillator(s). Future examination of seasonal aspects of these rhythms, may, however, cause this hypothesis to be modified.     

The role of the pineal and the retinae in the expression of circadian locomotor rhythmicity in the ruin lizard,Podarcis sicula

Summary A marked interspecific variability in the role played by the pineal and the retinae characterizes the circadian system of lizards. I examined the role played by these structures in a new model species, the ruin lizard, Podarcis sicula. In constant temperature and darkness pinealectomy as well as bilateral removal of the retinae produced significant changes (both lengthening and shortening) in the freerunning period of locomotor rhythms. Circadian activity time was also affected by pinealectomy. Circadian locomotor rhythmicity persisted in all cases even when both operations were combined in the same individuals. This demonstrates in Podarcis sicula the existence of an oscillatory system outside the pineal and the retinae which can drive locomotor rhythms. The period changes recorded after pinealectomy as well as after bilateral removal of the retinae specifically suggest that both the pineal and the retinae play a modulating role on circadian oscillators located elsewhere in the system, with the final effect of stabilizing the overt rhythms.Abbreviations DD constant darkness - LL constant light - PIN-X pinealectomy - RET-X bilateral removal of the retinae - SHAM sham pinealectomy - circadian activity time - freerunning circadian period     

Interseasonal variation in the circadian rhythms of locomotor activity and temperature selection in sleepy lizards, <Emphasis Type="Italic">Tiliqua rugosa</Emphasis>

Few studies in non-mammalian vertebrates have examined how various effectors of the circadian system interact. To determine if the daily locomotor and behavioural thermoregulatory rhythms of Tiliqua rugosa are both controlled by the circadian system in different seasons, lizards were tested in laboratory thermal gradients in four seasons and in constant darkness. Circadian rhythmicity for both rhythms was present in each season, being most pronounced in spring and summer and least evident in autumn. Most lizards displayed a unimodal locomotor activity pattern across all seasons. However, some individuals presented a bimodal locomotor activity pattern in spring and summer. Seasonal variations in the phase relationships of both rhythms to the light:dark (LD) cycle were demonstrated. No seasonal differences in the free-running period lengths of either rhythm were detected, raising the possibility that a single circadian pacemaker drives both rhythms in this species. Our present results demonstrate that both rhythms are similarly controlled by the circadian system in each season. Although seasonal variations in the thermal preferences of reptiles both in the field and laboratory have previously been well documented, this study is the first to demonstrate circadian rhythms of temperature selection in a reptile species in each season.     

Circadian locomotor rhythm masked by the female reproduction cycle in cockroaches

Blattella bisignata (Brunner) and B. germanica (L.) are oviparous cockroaches with cyclic reproductive behaviour, but in B. germanica only males show circadian rhythmicity of locomotion at 28°C and DD (constant darkness). In B. bisignata, males and virgin females cockroaches entrained by light–dark cycles show free‐running rhythmicity in DD, and most activities occur during the subjective night. Daily locomotor activities of virgin females show cyclic changes that coincided with ovarian development. Virgin females also exhibit calling behaviour during the subjective night, and this shows a free‐running rhythm. Male mate‐finding locomotion and female calling behaviour are under circadian control, so the timing for both behaviours is synchronized. However, most mated females do not show a locomotor free‐running rhythm under DD conditions. Our results indicate that only mated females could not express a circadian locomotor rhythm. Pregnancy reduces a female’s locomotory intensity and masks the expression of a circadian locomotor rhythm. We attribute the differences in circadian locomotory rhythms between these two species to their living environments and mate‐finding strategies.     

Pineal and retinal melatonin is involved in the control of circadian locomotor activity and body temperature rhythms in the pigeon

Summary Although pinealectomy or blinding resulted in loss of the clarity of the free-running rhythm of locomotor activity and body temperature and reduced the peak level of circulating melatonin rhythms to approximately a half in intact pigeons, neither pinealectomy nor blinding abolished any of these rhythms. However, when pinealectomy and blinding were combined, the rhythms of locomotor activity and body temperature disappeared in prolonged constant dim light, and melatonin concentration was reduced to the minimum level of detection. In order to examine the role of melatonin in the pigeon's circadian system, it was administered either daily or continuously to PX + EX-pigeons in LLdim. Daily administration of melatonin restored circadian rhythms of locomotor activity which entrained to melatonin injections, but continuous administration did not induce any remarkable change of locomotor activity. These results suggest that melatonin synthesized in the pineal body and the eye contributes to circulating melatonin and its rhythmicity is important for the control of circadian rhythms of locomotor activity and body temperature in the pigeon.Abbreviations LD Light-dark - LLdim constant dim light - LLbright constant bright light - PX pinealectomy - EX blinding - SCN suprachiasmatic nucleus     

Circadian Rhythms of Enzymes' Activity in Mice Tissues During Exposure to Continuous Illumination

Activity rhythms of enzymes were determined in various tissues of C57BL/6J male mice. The determinations were carried out on mice which were kept in 14 hr light: 10 hr dark regimen, and on day 2, day 5 and day 21 during exposure to continuous illumination. Locomotor activity rhythms were followed in light: dark and up to the seventh day in constant light. All the activities exhibited a significant circadian rhythm in the light: dark regimen. During the exposure to continuous illumination, the locomotor activity exhibit a free running circadian rhythm with a consistent 24 hr and 40 min, major period component. At the same time recording the rhythms of enzyme activity; enzymes exhibited various formats of response which differed from those of the locomotor activity. The results suggest that rhythms of enzyme activity, as well as the desynchronization of the rhythms, are not enzyme specific.     

Circadian locomotor rhythms in the desert iguana I. The role of the eyes and the pineal

Summary The pineal and the eyes are known to be important components in the circadian system of some species of lizards; their effects may be mediated by the hormone melatonin. We examined the role played by these structures in the desert iguana (Dipsosaurus dorsalis). Surgical removal of the pineal had no effect on circadian locomotor rhythms, even though this procedure abolished the circadian rhythm of melatonin in the blood. Furthermore, when the isolated pineal of Dipsosaurus was studied in organ culture, it showed no circadian rhythm of melatonin secretion, as do pineals of some other lizard species, although it did produce large quantities of this hormone. Bilateral ocular enucleation had only small effects on the freerunning period of locomotor rhythms, without affecting melatonin levels in the blood. Behavioral circadian rhythms persisted in desert iguanas subjected to both enucleation and pinealectomy. These data suggest that neither the pineal nor the eyes are central components of the circadian pacemaking system in Dipsosaurus, nor is melatonin critically involved in maintaining its organization.Abbreviations CT circadian time - ZT zeitgeber time - LL constant light - LD light-dark cycle - DD constant darkness - freerunning circadian period     

Period and phase control in a multioscillatory circadian system (Iguana iguana)

The circadian system of the lizard Iguana iguana is composed of several independent pacemakers that work in concert: the pineal gland, retinae of the lateral eyes, and a fourth oscillator presumed to be located in the hypothalamus. These pacemakers govern the circadian expression of multiple behaviors and physiological processes, including rhythms in locomotor activity, endogenous body temperature, electroretinogram, and melatonin synthesis. The numerous, easily measurable rhythmic outputs make the iguana an ideal organism for examining the contributions of individual oscillators and their interactions in governing the expression of overt circadian rhythms. The authors have examined the effects of pinealectomy and enucleation on the endogenous body temperature rhythm (BTR) and locomotor activity rhythm (LAR) of juvenile iguanas at constant temperature both in LD cycles and in constant darkness (DD). They measured the periods (tau) of the circadian rhythms of LAR and BTR, the phase relationships between them in DD (psiAT), and the phase relationship between each rhythm and the light cycle (psiRL). Pinealectomy lengthened tau of locomotor activity in all animals tested and abolished the BTR in two-thirds of the animals. In those animals in which the BTR did persist following pinealectomy, tau lengthened to the same extent as that of locomotor activity. Pinealectomy also delayed the onset of activity with respect to its normal phase relationship with body temperature in DD. Enucleation alone had no significant effect on tau of LAR or BTR; however, after enucleation, BTR became 180 degrees out of phase from LAR in DD. After both pinealectomy and enucleation, 4 of 16 animals became arrhythmic in both activity and body temperature. Their data suggest that rhythmicity, period, and phase of overt circadian behaviors are regulated through the combined output of multiple endogenous circadian oscillators.     

THERMOCYCLIC AND PHOTOCYCLIC ENTRAINMENT OF CIRCADIAN LOCOMOTOR ACTIVITY RHYTHMS IN SLEEPY LIZARDS,TILIQUA RUGOSA

Australian sleepy lizards (Tiliqua rugosa) exhibit marked locomotor activity rhythms in the field and laboratory. Light-dark (LD) and temperature cycles (TCs) are considered important for the entrainment of circadian locomotor activity rhythms and for mediating seasonal adjustments in aspects of these rhythms, such as phase, amplitude, and activity pattern. The relative importance of 24 h LD and TCs in entraining the circadian locomotor activity rhythm in T. rugosa was examined in three experiments. In the first experiment, lizards were held under LD 12:12 and subjected to either a TC of 33:15?°?C in phase with the LD cycle or a reversed TC positioned in antiphase to the LD cycle. Following LD 12:12, lizards were maintained under the same TCs but were subjected to DD. Activity was restricted to the thermophase in LD, irrespective of the lighting regime and during the period of DD that followed, suggesting entrainment by the TC. The amplitude of the TC was lowered by 8?°?C to reduce the intensity and possible masking effect of the TC zeitgeber in subsequent experiments. In the second experiment, lizards were held under LD 12.5:11.5 and subjected to one of three treatments: constant 30?°?C, normal TC (30:20?°?C) in phase with the LD cycle, or reversed TC. Following LD, all lizards were subjected to DD and constant 30?°?C. Post-entrainment free-run records revealed that LD cycles and TCs could both entrain the locomotor rhythms of T. rugosa. In LD, mean activity duration (α) of lizards in the normal TC group was considerably less than that in the constant 30?°?C group. Mean α also increased between LD and DD in lizards in the normal TC group. Although there was large variation in the phasing of the rhythm in relation to the LD cycle in reversed TC lizards, TCs presented in phase with the LD cycle most accurately synchronized the rhythm to the photocycle. In the third experiment, lizards were held in DD at constant 30?°?C before being subjected to a further period of DD and one of four treatments: normal TC (06:00 to 18:00 h thermophase), delayed TC (12:00 to 00:00 h thermophase), advanced TC (00:00 to 12:00 h thermophase), or control (no TC, constant 30?°?C). While control lizards continued to free-run in DD at constant temperature, the locomotor activity rhythms of lizards subjected to TCs rapidly entrained to TCs, whether or not the TC was phase advanced or delayed by 6 h. There was no difference in the phase relationships of lizard activity rhythms to the onset of the thermophase among the normal, delayed, and advanced TC groups, suggesting equally strong entrainment to the TC in each group. The results of this experiment excluded the possibility that masking effects were responsible for the locomotor activity responses of lizards to TCs. The three experiments demonstrated that TCs are important for entraining circadian locomotor activity rhythms of T. rugosa, even when photic cues are conflicting or absent, and that an interaction between LD cycles and TCs most accurately synchronizes this rhythm. (Author correspondence: david.j.ellis@adelaide.edu.au)     

Role of suprachiasmatic nuclei in circadian and light-entrained behavioral rhythms of lizards

To establish whether the suprachiasmatic nuclei (SCN) of the Ruin lizard (Podarcis sicula) play a role in entrainment of circadian rhythms to light, we examined the effects of exposure to 24-h light-dark (LD) cycles on the locomotor behavior of lizards with SCN lesions. Lizards became arrhythmic in response to complete SCN lesion under constant temperature and constant darkness (DD), and they remained arrhythmic after exposure to LD cycles. Remnants of SCN tissue in other lesioned lizards were sufficient to warrant entrainment to LD cycles. Hence, the SCN of Ruin lizards are essential both to maintain locomotor rhythmicity and to mediate entrainment of these rhythms to light. We also asked whether light causes expression of Fos-like immunoreactivity (Fos-LI) in the SCN. Under LD cycles, the SCN express a daily rhythm in Fos-LI. Because Fos-LI is undetectable in DD, the rhythm seen in LD cycles is caused by light. We further showed that unilateral SCN lesions in DD induce dramatic period changes. Altogether, the present data support the existence of a strong functional similarity between the SCN of lizards and the SCN of mammals.     

Differential effects of pinealectomy on circadian rhythms of feeding and perch hopping in the European starling

To study the effects of pinealectomy on the circadian rhythms of locomotor activity and feeding. European starlings (Sturnus vulgaris) were held in constant light (0.2 lux and 200 lux) and under constant temperature conditions. Locomotor activity was measured by means of perches with microswitches mounted underneath, and feeding with an infrared photocell system at the feeder. Pinealectomy consistently led to disturbances in perch-hopping rhythms and often to a complete loss of rhythmicity as revealed by periodogram analysis. In some birds, perch-hopping rhythms recovered following a period of initial arrhythmicity. When a perch-hopping rhythm was present, its period was usually shorter than it had been before pinealectomy. In contrast to its effects on perch hopping, pinealectomy had no effect on the persistence of feeding rhythmicity, although its period, like that of the hopping rhythm, decreased after this operation. These results support the hypothesis derived from previous studies that the circadian organization of feeding is different from that of perch hopping. Different circadian pacemakers may be involved, but other models may possibly explain the data just as well.     

Light/dark‐induced effects on behavioral rhythms in suprachiasmatic nucleus‐lesioned rats irrespective of the presence of functional suprachiasmatic nucleus brain implants

Summary

Suprachiasmatic nucleus (SCN)‐lesioned rats which had received a fetal SCN graft were kept in constant red light for three months. After this period it was examined whether those rats that showed a recovered free‐running circadian rhythm could be entrained to light/dark cycles. To this end, they were subjected to a 12 h light/12 h dark schedule, followed by a 12 h light shift and again to dark conditions. In addition, the same regime was imposed on SCN‐grafted rats without recovered circadian rhythms and on sham‐grafted animals with a lesion, which were studied as controls. The presence of an SCN graft was identified immunocytochemically by the presence of vasopressin, vasoactive intestinal polypeptide and somatostatin cells.

Drinking, eating and wheel‐running rhythms were found to synchronize to the light/dark cycles in all rats, not with standing the presence of an SCN graft was. A 12 h light shift was immediately followed by a shift in the three rhythms. Under final dark conditions, free‐running patterns reappeared in rhythm‐recovered animals, without any convincing evidence for entrainment of the rhythms in the pattern of transition.

Behavioral rhythms in SCN‐lesioned rats are apparently masked by 12 h light/dark schedules via other visual pathways than the direct projection from the retina to the SCN.     

Endogenous rhythms of locomotion in the American horseshoe crab, Limulus polyphemus

American horseshoe crabs (Limulus polyphemus) exhibit clear circadian rhythms of visual sensitivity in the laboratory and in the field they exhibit seasonal patterns of mating behavior that are closely associated with the tides. Recent reports suggest that Limulus locomotor activity may be controlled by endogenous circadian and/or circatidal clocks and that light:dark (LD) cycles may affect the rhythmic output of both of these clocks. In this study, we examined locomotor behavior in the laboratory to determine the extent of this endogenous activity and to examine the influence of LD cycles on these rhythms. Thirty-three L. polyphemus were captured during the breeding season and their activity was monitored with activity boxes and “running wheels” in seawater kept at constant temperature and salinity. Activity patterns were analyzed using visual inspection of actograms and Chi-square and Lomb-Scargle periodograms. Overall, 36% of the animals was significantly more active during L, while only 12% was more active during D (52% showed no preference). Circatidal rhythms were observed in LD in 67% of the horseshoe crabs. Surprisingly, LD cycles appeared to synchronize these rhythms at times. In DD, the majority of animals tested (63%) exhibited circatidal rhythms that persisted for at least seven days. Overall, the results demonstrate that an endogenously controlled tidal rhythm of locomotion operates during, and significantly after, the breeding season in this species. In addition, the present results are consistent with the presence of circalunidian oscillators controlling these rhythms.     

The Circadian System of Ruin Lizards: A Seasonally Changing Neuroendocrine Loop?

Previous studies have shown that the amplitude of daily melatonin production in cultured ruin lizard pineal organs explanted in the summer is significantly higher than that from organs explanted in the winter. To test whether seasonal photoperiodic changes are decoded autonomously by the pineal gland, pineals explanted in summer were cultured in vitro and exposed to changes between winter and summer photoperiods. The changes in photoperiod duration did not affect the daily profiles of in vitro melatonin production. The discrepancy between the present in vitro results and those from lizards exposed to winter or summer photoperiods before pineal explantation supports the view that circadian information entering the pineal gland via its innervation is involved in determining seasonal changes of melatonin production in ruin lizards. We further examined whether a central component of the circadian system of ruin lizards, specifically the retinae of the lateral eyes, expresses similar seasonal changes in function as does the pineal gland. We did not find any difference between summer and autumn‐winter in the effectiveness of either bilateral retinalectomy or optic nerve lesion—at the level of the optic chiasm—in altering circadian locomotor behavior in constant conditions. Both surgical procedures mostly induced a shortening of the free‐running period of the locomotor rhythm of similar magnitude in all seasons. Thus, the retinae do not appear to participate in the seasonal reorganization of the circadian system in ruin lizards.     

Seasonal variations in circadian rhythms of plasma melatonin in ruin lizards

We examined melatonin profiles of ruin lizards in different seasons (spring, summer, and autumn) under light:dark (LD) and concomitant responses when transferred to continuous darkness (DD) to determine the degree to which previously reported seasonally dependent effects of pinealectomy on locomotor behavior are related to melatonin secretion. The amplitude of the melatonin rhythm and the amount of melatonin produced over 24 h varied with season. In spring, the amount of melatonin produced was greatest and the amplitude 4- 5 times that found in summer or autumn. The degree of self-sustainment of the melatonin rhythm when transferred to DD also varied with season. In DD, melatonin levels remained high but did not exhibit circadian variation in spring. In summer, the melatonin profile persisted virtually unchanged in DD, showing the existence of a circadian rhythm. Finally, in the fall there was no circadian variation in DD and levels remained low. These responses correspond closely to previously reported effects of pinealectomy on locomotor behavior where there is little or no effect of pinealectomy in spring or fall but a profound alteration of locomotor behavior in summer. These results suggest that the seasonally dependent effects of pinealectomy on locomotor behavior in ruin lizards are related to a seasonally mediated change in the degree of self-sustainment of some component of the circadian pace-making system of which melatonin plays some role.     

Seasonal effects on the freerunning rhythm of circadian activity of longnose dace (Rhinichthys cataractae)

Synopsis Freerunning circadian rhythms of locomotor activity in individual longnose dace sampled from a population at 41°N latitude were recorded under constant darkness throughout the year. There was an annual cycle in the length of the circadian period, with maximum and minimum lengths of mean period of 23.6 and 21.6 h recorded during summer (June) and winter (December), respectively. These annual changes in period length may have resulted from seasonal changes in entrainment by natural light-dark cycles and their after-effects on endogenous circadian rhythms. The possibility of an endogenous circannual rhythm was also considered.     

Probing the circadian oscillator of a mammal by two-pulse perturbations

When organisms are maintained under constant conditions of light and temperature, their endogenous circadian rhythms free run, manifesting their intrinsic period. The phases of these free-running rhythms can be shifted by stimuli of light, temperature, and drugs. The change from one free-running steady state to another following a perturbation often involves several transient cycles (cycles of free-running rhythm drifting slowly to catch up with the postperturbation steady state). Although the investigation of oscillator kinetics in circadian rhythms of both insects and mammals has revealed that the circadian pacemaker phase shifts instantaneously, the phenomenon of transient cycles has remained an enigma. We probed the phases of the transient cycles in the locomotor activity rhythm of the field mouse Mus booduga, evoked by a single light pulse (LP), using LPs at critically timed phases. The results of our experiments indicate that the transient cycles generated during transition from one steady state to another steady state do not represent the state of the circadian pacemaker (basic oscillator) controlling the locomotor activity rhythm in Mus booduga. (Chronobiology International, 17(2), 129-136, 2000)