Effects of transient and continuous wheel running activity on the upper and lower limits of entrainment to light-dark cycles in female hamsters

The entrainment limits to light-dark cycles can be modified by the experimental conditions under which they are tested. Among the factors that may influence entrainment is the amount of wheel running exerted by the animal. In the present work, the effects of transitory and continuous wheel running on entrainment to light-dark cycles were tested using a range of T cycles at the entrainment limits. Four groups of female hamsters were submitted to 1 h stepwise changes in T cycles. Two groups were exposed to T cycles of which the period was shortened at the lower limit from T22 to T18, and the other two groups were exposed to cycles that lengthened at the upper limit from T27 to T32. One of the groups at the lower limit and one at the upper limit had continuous access to a running wheel, while the others had the wheel locked, except at certain T when a lack of period control by T cycle appeared. The study demonstrates that access to running wheel widens the limits of entrainment to LD cycles. Specifically, the following observations were made: the effects of wheel running for entrainment were more evident in the groups with continuous access to wheel, as they did entrain to T19 and T32; continuous access to a wheel produced aftereffects only after T19, but not under T32; and when animals without a wheel showed relative coordination, unlocking the wheel favored entrainment in all the animals at T31, but in only 1 out 6 at T19. All of these indicate a different effect of the wheel running on the upper and lower limits of entrainment.     

Effects of Transient and Continuous Wheel Running Activity on the Upper and Lower Limits of Entrainment to Light‐Dark Cycles in Female Hamsters

The entrainment limits to light‐dark cycles can be modified by the experimental conditions under which they are tested. Among the factors that may influence entrainment is the amount of wheel running exerted by the animal. In the present work, the effects of transitory and continuous wheel running on entrainment to light‐dark cycles were tested using a range of T cycles at the entrainment limits. Four groups of female hamsters were submitted to 1 h stepwise changes in T cycles. Two groups were exposed to T cycles of which the period was shortened at the lower limit from T22 to T18, and the other two groups were exposed to cycles that lengthened at the upper limit from T27 to T32. One of the groups at the lower limit and one at the upper limit had continuous access to a running wheel, while the others had the wheel locked, except at certain T when a lack of period control by T cycle appeared. The study demonstrates that access to running wheel widens the limits of entrainment to LD cycles. Specifically, the following observations were made: the effects of wheel running for entrainment were more evident in the groups with continuous access to wheel, as they did entrain to T19 and T32; continuous access to a wheel produced aftereffects only after T19, but not under T32; and when animals without a wheel showed relative coordination, unlocking the wheel favored entrainment in all the animals at T31, but in only 1 out 6 at T19. All of these indicate a different effect of the wheel running on the upper and lower limits of entrainment.     

The Role of Wheel Running in the Coupling of Two Simultaneous Circadian Rhythms of Motor Activity in the Rat

Motor activity is among the non-photic stimuli that act on the internal clock. We have tested the role of motor activity in the circadian pattern of rats under conditions near the lower limits of entrainment, that induce circadian rhythm dissociation. Three groups of 8 rats each were used: a) rats kept individually in 25×25×15 cm cages, b) rats in 50×25×15 cm cages, and c) rats in 50×25×15 cm cages with access to a running wheel. All the rats were kept under light-dark cycles of 22 hours (T22, 11L:11D) for 50 days, after which they were transferred to constant darkness. All the rats without a running wheel showed a motor activity pattern with two statistically significant circadian rhythms in the periodogram of Sokolove and Bushell: one circadian component entrained by the LD cycle, and another free-running. The rats with access to a running wheel showed several patterns: 5 rats showed only one rhythm entrained to the LD cycle, 2 rats showed circadian rhythm dissociation, and 1 showed only a free running rhythm. We believe that the simultaneous manifestation of two circadian components reflects the functional dissociation of the oscillators population that constitutes the circadian pacemaker, of the rat. Physical exercise acts on the pacemaker reinforcing the strongest group of oscillators, which, depending on the structure of the circadian system of the rat, is usually the one entrained to the LD cycle. This study supports the hypothesis that motor activity couples the oscillators that form the circadian system of the rat.     

The Role of Wheel Running in the Coupling of Two Simultaneous Circadian Rhythms of Motor Activity in the Rat

Motor activity is among the non-photic stimuli that act on the internal clock. We have tested the role of motor activity in the circadian pattern of rats under conditions near the lower limits of entrainment, that induce circadian rhythm dissociation. Three groups of 8 rats each were used: a) rats kept individually in 25×25×15 cm cages, b) rats in 50×25×15 cm cages, and c) rats in 50×25×15 cm cages with access to a running wheel. All the rats were kept under light-dark cycles of 22 hours (T22, 11L:11D) for 50 days, after which they were transferred to constant darkness. All the rats without a running wheel showed a motor activity pattern with two statistically significant circadian rhythms in the periodogram of Sokolove and Bushell: one circadian component entrained by the LD cycle, and another free-running. The rats with access to a running wheel showed several patterns: 5 rats showed only one rhythm entrained to the LD cycle, 2 rats showed circadian rhythm dissociation, and 1 showed only a free running rhythm. We believe that the simultaneous manifestation of two circadian components reflects the functional dissociation of the oscillators population that constitutes the circadian pacemaker, of the rat. Physical exercise acts on the pacemaker reinforcing the strongest group of oscillators, which, depending on the structure of the circadian system of the rat, is usually the one entrained to the LD cycle. This study supports the hypothesis that motor activity couples the oscillators that form the circadian system of the rat.     

History-dependent changes in entrainment of the activity rhythm in the Syrian hamster (Mesocricetus auratus)

The authors have studied the activity rhythm of Syrian hamsters exposed to square LD cycles with a 22-h period (T22) with the aim of testing the effects of the previous history on the rhythmic pattern. To do so, sequential changes of different lighting environments were established, followed by the same LD condition. Also, the protocol included T22 cycles with varying lighting contrasts to test the extent to which a computational model predicts experimental outcomes. At the beginning of the experiment, exposure to T22 with 300 lux and dim red light occurring respectively at photophase and scotophase (LD300/dim red) mainly generated relative coordination. Subsequent transfer to cycles with approximately 0.1-lux dim light during the scotophase (LD300/0.1) promoted entrainment to T22. However, a further reduction in light intensity to 10 lux during the photophase (LD10/0.1) generated weak and unstable T22 rhythms. When, after that, animals were transferred again to the initial LD300/dim red cycles, the amplitude of the rhythm still remained very low, and the phases were very unstable. Exposure to constant darkness partially restored the activity rhythm, and when, afterwards, the animals were submitted again to LD300/dim red cycles, a robust T22 rhythm appeared. The results demonstrate history-dependent changes in the hamster circadian system because the locomotor activity pattern under the same T22 cycle can show relative coordination or unstable or robust entrainment depending on the prior lighting condition. This suggests that the circadian system responds to environmental stimuli depending on its previous history. Moreover, computer simulations allow the authors to predict entrainment under LD300/0.1 cycles and indicate that most of the patterns observed in the animals due to the light in the scotophase can be explained by different degrees of coupling among the oscillators of the circadian system.     

Range of entrainment of rat circadian rhythms to sinusoidal light-intensity cycles

The range of entrainment of the circadian behavioral rhythm was compared between two groups of Sprague-Dawley rats (each n = 10) exposed to daily cycles of rectangular light-dark alternation (LD) and sinusoidal fluctuations of light intensity (SINE), respectively. The maximum illuminance (20 lx), the minimum illuminance (0.01 lx), and the total amount of light exposure per cycle were the same under the two lighting conditions. The periods (Ts) of both lighting cycles were lengthened stepwise from 24 through 25, 26, 26.5, 27, 27. 5, and 28 h to 28.5 h in experiment 1 and were shortened stepwise from 24 through 23.5, 23, and 22.5 h to 22 h in experiment 2. Each T cycle lasted for 30 cycles. In experiment 1, 60% of rats under the LD condition entrained up to T = 28.5 h, whereas 50% of rats under the SINE condition entrained up to T = 28.5 h. In experiment 2, no animal under the LD condition entrained to T < 23.5 h, whereas 40% of rats under the SINE condition entrained down to T = 23 h and 20% of rats remained to entrain down to T = 22 h cycles. The phase angle of entrainment was systematically changed, depending on T under both conditions. These results suggest that the lower limit of entrainment is expanded under the SINE condition compared with the LD condition.     

Effect of Behavioural Feedback on Circadian Clocks of the Nocturnal Field Mouse Mus booduga

The effect of 'novel running wheels' on circadian clocks of the nocturnal field mouse Mus booduga was investigated during free-running and entrained conditions. In order to find out whether daily access to novel running wheels can entrain the locomotor activity rhythms experimental animals (n = 6) were provided with 'novel running wheels' at a fixed time of the day. The control animals (n = 5) were handled similar to the experimental animals but were not given access to novel running wheels. The results show that daily access to novel running wheels entrained the free-running locomotor activity rhythm of these mice. The post-entrainment free-running period (τ) of the experimental animals was significantly shorter than the pre-entrainment τ, whereas the pre- and post-treatment τ of the control animals did not differ significantly. In separate set of experiments, the effect of access to novel running wheels on the rate of re-entrainment was studied after a 6 h phase advance/delay in 24 h (12:12 h) light/dark (LD) cycles. Experimental animals were given access to novel running wheels for 3-h, 1 h after the 'lights-off' only on the first day of the 'new LD cycles'. Experimental animals took fewer cycles to re-entrain to 6-h phase advanced LD cycles compared to the control animals. After a phase delay in the LD cycles by 6h, the experimental animals took more number of cycles to re-entrain compared to the control animals. These results thus suggest that access to novel running wheel can act as a Zeitgeber for the circadian clocks of the nocturnal mouse M. booduga, and can also modify the rates of re-entrainment to phase shifted LD cycles, in a time-dependent manner.     

Temperature entrainment of the circadian cuticle deposition rhythm in Drosophila melanogaster

The cuticle deposition rhythm, which is observed in the apodeme of the furca in the thorax, is controlled by a peripheral circadian clock in the epidermal cells and entrained to light-dark (LD) cycles via CRYPTOCHROME (CRY) in Drosophila melanogaster. In the present study, we examined the effects of temperature (TC) cycles and the combination of LD and TC cycles on entrainment of the cuticle deposition rhythm. The rhythm was entrained to TC cycles, whose period was 28 h. In T = 21 and 24 h, the rhythm was entrained to TC cycles in some individuals. CRY is not necessary for temperature entrainment of the cuticle deposition rhythm because the rhythm in cry(b) (lacking functional CRY) was entrained to TC cycles. Temperature entrainment of the rhythm was achieved even when the thoraxes or furcae were cultured in vitro, suggesting that the mechanism for temperature entrainment is independent of the central clock in the brain and the site of the thermoreception resides in the epidermal cells. When LD and TC cycles with different periods were applied, the rhythm was entrained to LD cycles with a slight influence of TC cycles. Thus, the LD cycle is a stronger zeitgeber than the TC cycle. The variance of the number of the cuticle layers decreased in the flies kept under LD and TC cycles with the same period in which the thermophase coincided with the photophase. Therefore, we conclude that LD and TC cycles synergistically entrain the rhythm. Synergistic effects of LD and TC cycles on entrainment were also observed even when the thoraxes were cultured in vitro, suggesting that the light and temperature information is integrated within the peripheral circadian system.     

History dependence of circadian pacemaker period in the cockroach

The freerunning period of circadian clocks in constant environmental conditions can be history-dependent, and one effect of entrainment of circadian clocks by light cycles is to cause long-lasting changes in the freerunning period that are termed after-effects. We have studied after-effects of entrainment to 22-h (LD 8:14) and 26-h (LD 8:18) light cycles in the cockroach Leucophaea maderae. We find that in cockroaches, the freerunning period of the locomotor activity rhythm, measured in constant darkness (DD), is 0.7h less after entrainment to T22 than after entrainment to T26. Induction of after-effects requires several days (>1 week) entrainment, and after induction, after-effects will persist in DD for over 40 days. Further after-effects are unaltered by phase-resetting of up to 12h caused by exposure to low-temperature pulses (7 degrees C) of 24 or 48h duration. After-effects also persist through re-entrainment for 2 weeks to 24-h light cycles. These results indicate that after-effects arise from stable changes in the circadian system that are likely to be independent of phase relationships among oscillators within the circadian system. We also show that entrainment to temperature cycles does not generate after-effects indicating that light may be unique in its ability to generate lasting changes in pacemaker period.     

Behavioral and neurochemical sources of variability of circadian period and phase: studies of circadian rhythms of npy-/- mice

The cycle length or period of the free-running rhythm is a key characteristic of circadian rhythms. In this study we verify prior reports that locomotor activity patterns and running wheel access can alter the circadian period, and we report that these treatments also increase variability of the circadian period between animals. We demonstrate that the loss of a neurochemical, neuropeptide Y (NPY), abolishes these influences and reduces the interindividual variability in clock period. These behavioral and environmental influences, from daily distribution of peak locomotor activity and from access to a running wheel, both act to push the mean circadian period to a value < 24 h. Magnitude of light-induced resetting is altered as well. When photoperiod was abruptly changed from a 18:6-h light-dark cycle (LD18:6) to LD6:18, mice deficient in NPY were slower to respond to the change in photoperiod by redistribution of their activity within the prolonged dark and eventually adopted a delayed phase angle of entrainment compared with controls. These results support the hypothesis that nonphotic influences on circadian period serve a useful function when animals must respond to abruptly changing photoperiods and point to the NPYergic pathway from the intergeniculate leaflet innervating the suprachiasmatic nucleus as a circuit mediating these effects.     

Entrainment of circadian activity rhythms in rats to melatonin administered at T cycles different from 24 hours

Entrainment of running wheel activity in DD was studied in adult male Long Evans rats exposed to cycles of a constant dose of melatonin (MEL; 100 microg/h) infused subcutaneously. The period (T) of the MEL cycle was initially kept at 24 h until stable entrainment was established; T was then changed in a stepwise manner, and each new T value was maintained for at least 20 cycles. Entrainment by phase advance occurred near circadian time 12 (activity onset), and the range of entrainment was between 30 and 35 min. The negative phase angle difference between activity onset and MEL onset increased as T values approached the entrainment limit, whereas no change in the duration of daily activity periods was found. No difference was observed between pre- and posttreatment values of the endogenous circadian period; hence, no aftereffects were found for any T value. These results indicate that the functional properties of entrainment to MEL are similar to those of entrainment to light, suggesting that both zeitgebers share a common timing mechanism.     

Effects of Photophase and Altitude on Oviposition Rhythm of the Himalayan Strains of Drosophila Ananassae

The effects of varying photophase and altitude of origin on the phase angle difference (Ψ) of the circadian rhythm of oviposition during entrainment to light‐dark (LD) cycles and the aftereffects of such photophases on the period of the free‐running rhythm (τ) in constant darkness (DD) were evaluated in two Himalayan strains of Drosophila ananassae, the high‐altitude (HA) strain from Badrinath (5,123 m above sea level=ASL) and the low‐altitude (LA) strain from Firozpur (179 m ASL). The Ψ (i.e., the hours from lights‐on of the LD cycle to oviposition median) of both strains was determined in LD cycles in which the photophase at 100 lux varied from 6 to 18 h/24 h. The HA strain was entrained by all LD cycles except the one with 6 h photophase in which it was weakly rhythmic, but the LA strain was entrained by only three LD cycles with photophases of 10, 12, and 14 h, but photophases of 6, 8, 16, and 18 h rendered it arrhythmic. Lights‐off transition of LD cycles was the phase‐determining signal for both strains as oviposition medians of the HA strain occurred～6 h prior to lights‐off, while those of the LA strain occurred～1 h after lights‐off. The Ψ of the HA strain increased from～2 h in 8 h photophase to～11 h in 18 h photophase, while that of the LA strain increased from～11 h in 10 h photophase to～15 h in 14 h photophase. The aftereffects of photophase of the prior entraining LD cycles on τ in DD were determined by transferring flies from LD cycles to DD. The τ of the HA strain increased from～19 to～25 h when transferred to DD from LD 8:16 and LD 18:6 cycles, respectively, whereas the τ of the LA strain increased from～26 to～28 h when transferred to DD from LD 10:14 and LD 14:10 cycles, respectively. Thus, these results demonstrate that the photophases of entraining LD cycles and the altitude of origin affected several parameters of entrainment and the period of the free‐running rhythm of these strains.     

Twilights widen the range of photic entrainment in hamsters

The range of entrainment of the circadian rhythm of locomotor activity was compared in four groups of Syrian hamsters (eight animals per group) initially exposed to daily light-dark (LD) cycles with either abrupt transitions between light and darkness (LD-rectangular) or simulated twilights (LD-twilight). Lighting was provided by arrays of white light-emitting diodes; daytime illuminance (10 lux) and the total amount of light emitted per day were the same in the two conditions. The period (T) of the LD cycles was then gradually increased to 26.5 h or gradually decreased to 21.5 h, at the rate of 5 min/day. Under LD-rectangular, the upper and lower limits of entrainment were 25.0 to 25.5 h and 22.0 to 22.5 h, respectively, whereas under LD-twilight, 50% of the animals exposed to the lengthening cycles were still entrained at T = 26.5 h and 50% of those exposed to the shortening cycles were still entrained at T = 21.5 h. In a second experiment, two groups of hamsters were exposed to fixed T = 25 h LD-rectangular (n = 15) or LD-twilight cycles (n = 7). Only 33% of the animals entrained in LD-rectangular, whereas 86% of the animals entrained in LD-twilight. Free-running periods in constant darkness were longer following successful entrainment to T = 25 h but did not differ between the animals that entrained to LD-rectangular and those that entrained to LD-twilight. The widening of the range of entrainment observed in LD-twilight indicates that twilight transitions increase the strength of the LD zeitgeber. In LD-twilight, successful entrainment to T = 26.5 h was accompanied by an expansion of activity time to 16.52+/-1.22 h, with activity onsets preceding mid-dusk by 12.56+/-2.15 h. Together with earlier data showing similar phase response curves for hour-long dawn, dusk, and rectangular light pulses, these results suggest that the effect of twilights on the range of entrainment may involve parametric rather than nonparametric mechanisms.     

Behavioral feedback regulation of circadian rhythm phase angle in light-dark entrained mice

Induced and spontaneous wheel running can alter the phase and period (tau) of circadian rhythms in rodents. The relationship between spontaneous running and the phase angle (psi) of entrainment to 24-h light-dark (LD) cycles was evaluated in C57BL/6j mice. With a wheel freely available, psi was significantly correlated with the absolute (r = 0.32) and relative (r = 0.44) amount of activity during the first 2 h of the activity period. When wheels were locked during the first half of the night in LD and then unlocked in constant dark (DD), mice exhibited a delayed psi and lengthened tau compared with mice that had wheels locked during the second half of the night. In DD, tau correlated negatively with total daily activity. To evaluate if wheel running modulates the phase-resetting actions of LD, phase shifts to light pulses were measured at two time points in DD, when daily activity levels differed by 40%. Phase delays to light were 56% greater when activity levels were lower. However, in a counterbalanced follow-up experiment, phase advances and delays to light pulses were not affected by the availability of wheels, although an effect of time in DD was replicated. Spontaneous activity can regulate psi and tau without altering the response of the pacemaker to light.     

Circadian rhythm of locomotor activity in the Japanese honeybee, Apis cerana japonica

Abstract.  To reveal circadian characteristics and entrainment mechanisms in the Japanese honeybee Apis cerana japonica , the locomotor-activity rhythm of foragers is investigated under programmed light and temperature conditions. After entrainment to an LD 12 : 12 h photoperiodic regime, free-running rhythms are released in constant dark (DD) or light (LL) conditions with different free-running periods. Under the LD 12 : 12 h regime, activity offset occurs approximately 0.4 h after lights-off transition, assigned to circadian time (Ct) 12.4 h. The phase of activity onset, peak and offset, and activity duration depends on the photoperiodic regimes. The circadian rhythm can be entrained to a 24-h period by exposure to submultiple cycles of LD 6 : 6 h, as if the locomotive rhythm is entrained to LD 18 : 6 h. Phase shifts of delay and advance are observed when perturbing single light pulses are presented during free-running under DD conditions. Temperature compensation of the free-running period is demonstrated under DD and LL conditions. Steady-state entrainment of the locomotor rhythm is achieved with square-wave temperature cycles of 10 °C amplitude, but a 5 °C amplitude fails to entrain.     

Tidal,Daily, and Lunar‐Day Activity Cycles in the Marine Polychaete Nereis virens

The burrow emergence activity of the wild caught ragworm Nereis virens Sars associated with food prospecting was investigated under various photoperiodic (LD) and simulated tidal cycles (STC) using a laboratory based actograph. Just over half (57%) of the animals under LD with STC displayed significant tidal (～12.4 h) and/or lunar‐day (～24.8 h) activity patterns. Under constant light (LL) plus a STC, 25% of all animals were tidal, while one animal responded with a circadian (24.2 h) activity rhythm suggestive of cross‐modal entrainment where the environmental stimulus of one period entrains rhythmic behavior of a different period. All peaks of activity under a STC, apart from that of the individual cross‐modal entrainment case, coincided with the period of tank flooding. Under only LD without a STC, 49% of the animals showed nocturnal (～24 h) activity. When animals were maintained under free‐running LL conditions, 15% displayed significant rhythmicity with circatidal and circadian/circalunidian periodicities. Although activity cycles in N. virens at the population level are robust, at the individual level they are particularly labile, suggesting complex biological clock‐control with multiple clock output pathways.     

Temporal flexibility in activity rhythms of a diurnal rodent,the ice rat (Otomys sloggetti)

ABSTRACT

Diurnality in rodents is relatively rare and occurs primarily in areas with low nighttime temperatures such as at high altitudes and desert areas. However, many factors can influence temporal activity rhythms of animals, both in the field and the laboratory. The temporal activity patterns of the diurnal ice rat were investigated in the laboratory with, and without, access to running wheels, and in constant conditions with running wheels. Ice rats appeared to be fundamentally diurnal but used their running wheels during the night. In constant conditions, general activity remained predominantly diurnal while wheel running was either nocturnal or diurnal. In some animals, entrainment of the wheel running rhythm was evident, as demonstrated by free-running periods that were different from 24 h. In other animals, the wheel running activity abruptly switched from nocturnal to subjective day as soon as the animals entered DD, and reverted back to nocturnal once returned to LD, suggesting the rhythms were masked by light. Wheel running rhythms appears to be less robust and more affected by light compared to general activity rhythms. In view of present and future environmental changes, the existence of more unstable activity rhythms that can readily switch between temporal niches might be crucial for the survival of the species.     

Circadian rhythms in the blowfly, Phormia terraenovae: control of phase within the range of entrainment

ABSTRACT Locomotor activity of individual blowflies, Phormia (= Protophormia) terraenovae R.D. (Diptera, Calliphoridae) was recorded by means of running wheels. A few days after emergence, adult flies were placed in the wheels and exposed to at least two of four light-dark cycles (LD) differing in cycle duration T (LD 11:11, 12:12, 13:13 and 14:14 h). The intensity of illumination was 400 lux in L and 2 lux in D. From the actograms, phase-angle differences were read off between onset of activity and light-on (ψ/_onset), and between end of activity and light-off (ψend)- Within the range of entrainment, ψ changed systematically from negative values in T =22 h to positive values in T =28 h: the mean change in ψ per hour change in T , expressed in degree of the full circadian cycles, was 20^o. Standard deviations of ψ around its mean were computed for ten-cycle intervals; in ψ_onset and in ψ_end standard deviation was minimal when 4ψ was close to zero, and increased steadily with increasing negative or positive ψ -values.     

Effects of photoperiod on rat motor activity rhythm at the lower limit of entrainment

The experiment described here studied the rat motor activity pattern as a function of the photoperiod of circadian light-dark cycles in the limits of entrainment (22-and 23-h periods). In most cases, the overt rhythm showed 2 circadian components: 1 that followed the external LD cycle and a 2nd rhythm that was free run. The expression of these components was directly dependent on the photoperiod, and there was a gradual transition in the manifestation of 1 or the other. The component with a period equal to that of the external cycle was more manifested under long photoperiods, while the other 1 was more expressed during short photoperiods. Also, the period of the free-running component was longer under T22 than T23. For each period, the free-running component was longer under a longer photoperiod. At first sight, the presence of these 2 components in most of the rats might appear to be due to the fact that in the limits of entrainment, some rats do not entrain and thus show a free-running rhythm plus masking. However, the gradation observed in the different patterns of the overt motor activity rhythm, especially those patterns related to the different balance between the 2 components and the length of the period of the free-running component under LD as a function of the photoperiod, suggests that the circadian system can be functionally dissociated.