Dissociation between the circadian rhythm of locomotor activity and the pineal clock in the Japanese newt

The circadian locomotor activity rhythm of the Japanese newt has been thought to be driven by a putative brain oscillator(s) subordinate to the pineal clock. The existence of mutual coupling between the pineal clock and the brain oscillator(s) in vivo was examined. We covered the newt's skull with aluminum foil and simultaneously reversed the light-dark cycle, thereby allowing the pineal organ to be exposed to constant darkness while the rest of the animal was exposed to the reversed light-dark cycle. In control animals, whose heads were covered with transparent plastic, the rhythm of synaptic ribbon number in the pineal photoreceptor cells was entrained to the reversed light-dark cycle. Rhythms from newts whose heads were shielded, however, were similar to those observed in the unoperated newts kept under constant darkness. The locomotor activity rhythms of both head-covered animals and control animals were entrained to the reversed light-dark cycle. These data suggest that extrapineal photoreception can entrain the putative brain oscillator(s), but not the pineal clock. Thus, at least in an aspect of photic entrainment, there seems to be little or no mutual coupling between the pineal clock and the putative brain oscillator(s) in the circadian system of the Japanese newt.Abbreviations LD light-dark - DD constant darkness - SCN suprachiasmatic nucleus - SR synaptic ribbon     

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     

Complete suprachiasmatic lesions eliminate circadian rhythmicity of body temperature and locomotor activity in golden hamsters

The effects of suprachiasmatic and control lesions on the circadian rhythms of locomotor activity and body temperature were studied in golden hamsters (Mesocricetus auratus) maintained in constant light as well as constant darkness. Large suprachiasmatic lesions, but not control lesions, eliminated circadian rhythmicity in locomotor activity as well as in body temperature. Analysis of the robustness of the rhythms of locomotor activity and body temperature in unlesioned and lesioned animals suggests that, because body temperature rhythmicity is more robust than locomotor rhythmicity, lesions that spare a small number of suprachiasmatic cells might abolish the latter but not the former. Our results do not support the hypothesis that the body temperature rhythm is controlled by a circadian pacemaker distinct from the main pacemaker located in the suprachiasmatic nuclei.     

Pharmacological effects of vinorelbine on body temperature and locomotor activity circadian rhythms in mice

The effects of vinorelbine (VRL) on the circadian rhythms in body temperature and locomotor activity were investigated in unrestrained B6D2F₁ mice implanted with radio-telemetry transmitters. A single intravenous VRL dose (24 or 12 mg/kg) was given at 7 h after light onset (HALO), a time of high VRL toxicity, and resulted in transient suppression of temperature and activity circadian rhythms in mice kept in light-dark (LD) 12h:12h. Such suppression was dose-dependent. It occurred within 1-5 d after VRL dosing. Recovery of both rhythms was partially complete within 5 d following the high dose and within 2 or 3 d after the low dose and was not influenced by suppression of photoperiodic synchronization by housing in continuous darkness. Moreover, VRL induced a dose-dependent relative decrease in amplitude and phase shift of the temperature circadian rhythm. The mesor and amplitude of the activity rhythm were markedly reduced following the VRL administration. The relevance of VRL dosing time was studied in mice housed in LD 12h:12h. Vinorelbine was injected weekly (20 mg/kg/injection) for 3 wk at 6 or 18 HALO. Vinorelbine treatment ablated the rest-activity and temperature rhythms 3-6 d after each dose, with fewer alterations after VRL dosing at 18 HALO compared to 6 HALO, especially for the body temperature rhythm. There was at least partial recovery 1 wk after dosing, suggesting the weekly schedule of drug treatment is acceptable for therapeutic purposes. Our findings demonstrate that VRL can transiently, yet profoundly, alter circadian clock function. Vinorelbine-induced circadian dysfunction may contribute to the toxicokinetics of this and possibly other anticancer drugs.     

Seasonality of circadian locomotor activity in an insect

Daily periodic locomotor activity of Carabus auronitens was recorded in a climate-constant laboratory with the animals exposed to naturally changing photoperiods. Most actograms exhibit directed seasonal variations of duration and phase position of daily activity. Seasonal locomotor activity starts in early spring (following dormancy) on a low daily level, first being confined to a short time span around dusk (and even shorter around dawn). In the course of season, the daily onsets of activity become closely related with sunset and the duration of daily activity is steadily extended with both parts of the bimodal phase fusing to a common, unimodal activity band by late spring. Subsequently, it is further extended into forenoon, until in summer (shortly before aestivation), spontaneous phase inversion turns activity periodicity from nocturnality into diurnality within 1 day. Such seasonal variations are paralleled by changes in the precision of synchronization of the individuals' activity rhythms to the entraining light/dark cycle. No geographical differences were detected. The results support the idea of the circadian clock as a system of two dynamically coupled physiological oscillators that invert their phase relation as soon as the natural dark phase falls short of some minimum-tolerable night length.Abbreviations LD light/dark cycle - nLD natural light/dark cycle - DD constant darkness - endogenous period length - SS sunset - SR sunrise - PRC phase-response curve     

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     

Light and temperature cycles as zeitgebers of zebrafish (Danio rerio) circadian activity rhythms

Light and temperature cycles are the most important synchronizers of biological rhythms in nature. However, the relative importance of each, especially when they are not in phase, has been poorly studied. The aim of this study was to analyze the entrainment of daily locomotor activity to light and/or temperature cycles in zebrafish. Under two constant temperatures (20°C and 26°C) and 12:12 light-dark (LD) cycles, zebrafish were most active during the day (light) time and showed higher total activity at the warmer temperature, while diurnalism was higher at 20°C than at 26°C (87% and 77%, respectively). Under thermocycles (12:12 LD, 26:20°C thermophase:chryophase or TC), zebrafish daily activity synchronized to the light phase, both when the thermophase and light phase were in phase (LD/TC) or in antiphase (LD/CT). Under constant dim light (3 lux), nearly all zebrafish synchronized to thermocycles (τ=24 h), although activity rhythms (60% to 67% of activity occurred during the thermophase) were not as marked as those observed under the LD cycle. Under constant dim light of 3 lux and constant temperature (22.5°C), 4 of 6 groups of zebrafish previously entrained to thermocycles displayed free-running rhythms (τ=22.9 to 23.6 h). These results indicate that temperature cycles alone can also entrain zebrafish locomotor activity.     

Effects of dark-light step transitions on circadian locomotor activity rhythms of the hagfish,Eptatretus burgeri

Captive hagfish,Eptatretus burgeri, were subjected to step transitions from continuous dark (DD) to continuous light (LL) and their locomotor activity patterns recorded. Free-running activity rhythms occurred in both the DD and LL regimes. The timing of the transition influenced the circadian period (τ), the relationship between the period response (δτ, or τ{inLL}–τ{inDD}, change in the circadian period) and the former being represented by a cosine curve δτ became most positive and most negative when hagfish underwent a DD-LL transition shortly before the beginning of the duration of activity (α) and shortly after the end of such, respectively. The phase response (δ-phase, change in the relative timing of the activity phase) was characterized by a delay in the activity phase after the DD-LL transition, although its magnitude bore no relation to the timing of the transition.     

Different circadian pacemakers control feeding and locomotor activity rhythms in European starlings

Summary In higher organisms, many physiological and behavioral functions exhibit daily variations, generated by endogenous circadian oscillators. It is not yet clear whether all the various rhythms that occur within an individual depend on one and the same pacemaker or whether different pacemakers are involved. To examine this question, the feeding and perch-hopping rhythms were measured in European starlings (Sturnus vulgaris) under light-dark cycles and continuous dim light. In dim light, the internal phase relationship between the feeding and perch-hopping rhythms changed systematically as a function of the circadian period, and the two rhythms could even dissociate and show different circadian periods in individuals with extremely long or extremely short circadian periods. Moreover, in some birds kept on lowamplitude light-dark cycles, the rhythm of feeding was synchronized 180° out of phase with the rhythm of locomotor activity. These results strongly suggest that in the European starling the feeding and locomotor activity rhythms are controlled by separate circadian pacemakers.     

A locus for circadian period of locomotor activity on mouse proximal chromosome 3

Lengthened circadian period of locomotor activity is a characteristic of a congenic strain of mice carrying a nonsense mutation in exon 5 of the carbonic anhydrase II gene, car2. The null mutation in car2 is located on a DBA/2J inbred strain insert on proximal chromosome 3, on an otherwise C57BL/6J genomic background. Since reducing the size of the congenic region would narrow the possible candidate genes for period, two recombinant congenic strains (R1 and R2) were developed from the original congenic strain. These new congenic strains were assessed for period, genetic composition, and the presence of immunoreactive carbonic anhydrase II. R1 mice were homozygous DBA/2J for the distal portion of the original DBA/2J insert, while R2 mice were homozygous DBA/2J for the proximal portion. R1 mice had a significantly lengthened period compared to R2 mice and wild-type C57BL/6J mice, indicating that the gene(s) affecting period is likely found within the reduced DBA/2J insert (~1 cM) in the R1 mice. The R1 mice also possessed the null mutation in car2. This study confirmed the presence of a gene(s) affecting period on proximal chromosome 3 and significantly reduced the size of the congenic region and the number of candidate genes. Future studies will focus on identifying the gene influencing period.     

Entrainment to light of circadian activity rhythms in tench (Tinca tinca)

The present article analyzes locomotor activity rhythms in Tinca tinca. To that end, three different experiments were conducted on 24 animals (20 g body weight) kept in pairs in 60-liter aquaria fitted with infrared sensors connected to a computer to continuously record fish movements. The first experiment was designed to study the endogenous circadian clock under free-running conditions [ultradian 40:40 min LD pulses and constant dark (DD)] and after shifting the LD cycle. Our results demonstrate that tench has a strictly nocturnal activity pattern, an endogenous rhythm being evident in 45.8% of the fish analyzed. The second experiment was conducted to test the influence of different photoperiods (LD 6:18, 12:12, 18:6, and 22:2) on locomotor activity, the results showing that even under an extremely long photoperiod, tench activity is restricted to dark hours. The third experiment examined the effect of light intensity on locomotor activity rhythms. When fish were exposed to decreasing light intensities (from 300:0 lux to 30:0, 3:0, and 0.3:0 lux) while maintaining a constant photoperiod (LD 12:12), the highest percentage of locomotor activity was in all cases associated with the hours of complete darkness (0 lux). In short, our results clearly show that (a) tench is a species with a strictly nocturnal behavior, and (b) daily activity rhythms gradually entrain after shifting the LD cycle and persist under free-running conditions, pointing to their circadian nature. However, light strongly influences activity rhythms, since (c) the length of the active phase is directly controlled by the photophase, and (d) strictly nocturnal behavior persists even under very dim light conditions (0.3 lux). The above findings deepen our knowledge of tench behavior, which may help to optimize the aquacultural management of this species, for example, by adjusting feeding strategies to their nocturnal behavior.     

Masking of locomotor activity in hamsters

The inhibition of locomotion by light (masking) was investigated in Syrian hamsters. When 1-h pulses of light were presented in the early night, activity was strongly suppressed by irradiances of about 1 lx or greater. Ultradian light-dark cycles were used as another way to study masking. Hamsters were unable to entrain to 3.5:3.5-h light-dark cycles, thus permitting the masking and the entraining effects of light to be distinguished. Light had greater suppressive effects on activity in home cages than on activity in novel running wheels. Moreover, in home cages activity remained very low for about 30 min after lights were turned off. Post-pulse suppression of activity was not simply a consequence of reduced running, as shown by experiments in which running was temporarily prevented by locking the wheels. A phase response curve for masking was obtained by placing hamsters in novel wheels for 3-h periods at various times throughout their circadian cycles, and then superimposing a 30-min light pulse. The suppressive effect of light was maximal around the onset of activity, which normally coincides with dusk in hamsters. This may have adaptive value in limiting foraging to the hours of darkness. Accepted: 8 February 1999     

Irradiance dependency of UV-A induced phase shifts in the locomotor activity rhythm of the field mouse Mus booduga

In the nocturnal field mouse Mus booduga, the responsiveness of the circadian system to UV-A light of 2.5 W/m² and 30 minutes duration is known to be phase dependent. The results of our experiments indicate that the phase shifts evoked by UV-A at the two phases, CT14 (circadian time 14) and CT20 increases nonlinearly with irradiance. (Chronobiology International, 17(6), 777-782, 2000)     

Feeding entrainment of locomotor activity rhythms in the goldfish is mediated by a feeding-entrainable circadian oscillator

Periodic food availability can act as a potent zeitgeber capable of synchronizing many biological rhythms in fishes, including locomotor activity rhythms. In the present paper we investigated entrainment of locomotor rhythms to scheduled feeding under different light and feeding regimes. In experiment 1, fish were exposed to a 12:12?h light/dark cycle and fed one single daily meal in the middle of the light phase. In experiment 2, we tested the effect of random versus scheduled feeding on the daily distribution of activity. During random feeding, meals were randomly scheduled with intervals ranging from 12 to 36?h, while scheduled feeding consisted of one single daily meal set in the middle of the light or dark phase. Finally, in experiment 3, we studied the synchronization of activity rhythms to feeding under constant darkness (DD) and after shifting the feeding cycle by either advancing or delaying the feeding cycle by 9?h. The results revealed that goldfish synchronized to feeding, overcame light entrainment and significantly changed their daily distribution of activity according to their feeding schedule. In addition, the daily activity pattern modulated by feeding differed between layers: a peak of activity being noticeable directly after feeding at the bottom, while an anticipatory behaviour was obvious at the surface of the tank. Under DD and no food, free-running rhythms averaging 25.5?± 1.9?h (mean?±?SD) were detected. In conclusion, some properties of feeding entrainment (e.g. anticipation of the feeding time, free-running rhythms following termination of periodic feeding, and the stability of ø after shifting the feeding cycle) suggested that goldfish have (a) separate but tightly coupled light- and food-entrainable oscillators, or (b) a single oscillator that is entrainable by both light and food (one synchronizer being eventually stronger than the other).     

Circadian rhythm of locomotor activity in individual workers of the wood ant Formica rufa

ABSTRACT. Individual worker ants isolated in an actograph exhibit circadian rhythms of locomotor activity. Entrainment occurs more readily in LD 18:6 h than in LD 12:12 h. The ants are either light-active or dark-active. Phase angle and duration of activity is influenced by photoperiod.     

Parametric and nonparametric entrainment of circadian locomotor rhythm in the Japanese honeybee Apis cerana japonica

The circadian rhythm of locomotor activity in the Japanese honeybee Apis cerana japonica was studied to determine the involvement of parametric and/or nonparametric entrainment. The rhythm was entrained to a skeleton photoperiod in which a 1-h first light pulse was imposed in the morning along with a second light pulse in the evening, as well as to a complete photoperiodic regime (LD 12:12). However, the timing of peak activity relative to the lights-off in the evening in the skeleton photoperiod was earlier than that in the complete photoperiod. A single daily light pulse in the evening entrained the rhythm, whereas a daily light pulse in the morning allowed free-running as in constant darkness. The free-running period (τ) of locomotor activity in constant light became longer as the light intensity increased. A Winfree's type I phase response curve of the locomotor activity rhythm was obtained using a single 1-h light pulse. The results suggest that both parametric and nonparametric entrainment are involved in the circadian rhythm of individual locomotor activity in this honeybee.     

Dissociation of photoreceptor cells from the pineal organ of the lamprey,Lampetra japonica

Summary Photoreceptor cells, nerve cells and supporting cells were dissociated from the pineal organ of the river lamprey, Lampetra japonica, by the use of 10 U/ml papain solution at 28° C for 20 min, followed by repeated trituration. With the aid of Nomarski interferencecontrast optics, photoreceptor cells, nerve cells and supporting cells were readily identified. Electron-microscopic examination revealed that isolated photoreceptor cells display an outer segment endowed with a few lamellar disks and connected to the inner segment (ellipsoid) via a connecting cilium. The structural features of the dissociated photoreceptor and supporting cells strongly resemble the morphology of the respective cellular elements in situ. We succeeded in culturing dissociated cells for time periods up to 48 h when the procedure described in detail was applied.     

Diazepam affects both level and amplitude of rat locomotor activity rhythm but has no effect on core body temperature

The present study investigates the effects of a chronic administration of diazepam, a benzodiazepine widely used as an anxiolytic, on locomotor activity and body core temperature rhythms in male Wistar rats housed under 12:12 light:dark (LD) cycle conditions. Diazepam was administered subcutaneously for 3 wks in a dosage of 3 mg/kg body weight/day, 1 h before the onset of darkness. Diazepam increased the level of locomotor activity from the first day until the end of treatment, and also increased the amplitude of the activity circadian rhythm, but only on the third wk of treatment. Diazepam exerted no effects on the length of the period and did not affect the phase of the locomotor activity rhythm. The body temperature rhythm of rats was affected neither by short-term (a single injection) nor by long-term (every day for 3 wks) diazepam treatment. Diazepam lacked effect on body core temperature even on the first day of administration, thereby ruling out the possibility of drug tolerance development. The fact that diazepam affects locomotor activity, but not core body temperature, suggests that different mechanisms mediate the actions of diazepam on locomotor activity and on core body temperature.     

Effect of ambient temperature on the circadian activity rhythm in common marmosets, Callithrix j. jacchus (primates)

Whereas the (zeitgeber) effect of ambient temperature T_a and temperature cycles TaC's on circadian rhythmicity has been well documented for heterofhermic mammals, inconsistent results have been obtained for strictly homeothermic species. Hence, it might be inferred that the susceptibility of the mammalian circadian timing system (CTS) to Ta and TaC's depends on the range of the animals' core and/or brain temperature rhythm. This hypothesis was tested in the common marmoset (Callithrix j. jacchus, n=12), a small diurnal primate with an amplitude in body temperature rhythm that is larger than for other homeothermic primates studied so far. Within the range 20-30°C, no systematic effects of constant Ta on most parameters of the marmosets' light-dark (LD)-entrained and free-running circadian activity rhythm (CAR) were found. Significant differences could be established in the average amount of activity per circadian cycle. It was highest at Ta 25°C (LD) and 20°C (light-light, LL) and most probably reflected a temperature-induced masking effect. A 24h trapezoidal TaC of 20:30°C entrained the free-running CAR in two of six marmosets and produced relative coordination in all others. Accordingly, in all animals tested, it had an effect on the CTS. In marmosets free running in LL at a Ta of 20°C or 30°C, 3h warm and cold pulses of 30°C and 20°C, respectively, produced neither systematic phase responses nor period responses of the CAR. So, there is no evidence of a phase-response mechanism underlying circadian entrainment. The results show that large-amplitude TaC's function as a weak zeitgeber for the marmosets' CTS. Since this zeitgeber effect is significantly larger than that found in owl monkeys, the results are consistent with the starting hypothesis that the zeitgeber effect of a given T,C on the mammalian CTS may be related to the amplitude of the species' core and/or brain temperature cycle.