Trinitat Cambras

Circadian desynchronization of core body temperature and sleep stages in the rat

Proper functioning of the human circadian timing system is crucial to physical and mental health. Much of what we know about this system is based on experimental protocols that induce the desynchronization of behavioral and physiological rhythms within individual subjects, but the neural (or extraneural) substrates for such desynchronization are unknown. We have developed an animal model of human internal desynchrony in which rats are exposed to artificially short (22-h) light–dark cycles. Under these conditions, locomotor activity, sleep–wake, and slow-wave sleep (SWS) exhibit two rhythms within individual animals, one entrained to the 22-h light–dark cycle and the other free-running with a period >24 h (τ>24 h). Whereas core body temperature showed two rhythms as well, further analysis indicates this variable oscillates more according to the τ>24 h rhythm than to the 22-h rhythm, and that this oscillation is due to an activity-independent circadian regulation. Paradoxical sleep (PS), on the other hand, shows only one free-running rhythm. Our results show that, similarly to humans, (i) circadian rhythms can be internally dissociated in a controlled and predictable manner in the rat and (ii) the circadian rhythms of sleep–wake and SWS can be desynchronized from the rhythms of PS and core body temperature within individual animals. This model now allows for a deeper understanding of the human timekeeping mechanism, for testing potential therapies for circadian dysrhythmias, and for studying the biology of PS and SWS states in a neurologically intact model.

Evolution of rat motor activity circadian rhythm under three different light patterns

The effects of the light pattern on the evolution of circadian motor activity rhythm of rats were studied in this work. Three different light patterns were used: LL (bright light, 300 lux), DD (dim red light) and LD (12:12 cycles). The animals used for the experiment were born and kept under each condition. At the day of weaning (21-22 days old) animals were isolated and their motor activity was detected by means of an inductive system. Data were recorded every 30 minutes for the first month after weaning. Periodogram analysis was applied to each animals data and the daily power spectra were calculated on the basis of the endogenous period, tau. The evolution of the rhythm was studied by examining the changes of the whole power spectra of the motor activity function, obtained by means of a Fourier analysis, through time. The power content of the circadian harmonic (PCCH) was considered to be a measure of the circadian character of the function. Results showed the predominancy of ultradian harmonics, when the animals were young, especially in LL, and the increase of the PCCH through time in all cases. Animals under LL show the circadian harmonic to be the main harmonic of the spectra at about day 15 after weaning, while the animals under DD show this harmonic as the main one from the first day. However, the power content of this harmonic increased until day 10. LD animals also showed the first harmonic as the main one from the time of weaning increasing the PCCH until day 7. These results are explained in respect to a multioscillatory system.(ABSTRACT TRUNCATED AT 250 WORDS)

Entrainment of the rat motor activity rhythm: effects of the light–dark cycle and physical exercise

The circadian system is believed to be composed of a population of oscillators that couple together and generate a single rhythm. If this coupling is not strong enough, the circadian system can be dissociated into two or more groups of oscillators, and this is manifested in a dissociation of the overt rhythm into at least two circadian components. This study aims to examine the influence of factors, such as the difference in impact between T and tau, light intensity, and access to a running wheel, on the distribution of motor activity throughout the light-dark (LD) cycle and the dissociation of the rhythm. Rats were submitted to LD cycles of 23 h (T23) or 25 h. For each such cycle, half the rats were submitted to high light intensity and the other half to low light intensity. For each of these conditions, half the rats were kept in small cages, and the other half were in cages with a running wheel. Rats were maintained first under LD cycles and afterwards under constant darkness (DD). Motor activity was recorded throughout the whole experiment by means of activity meters with infrared beams. Results show that the distribution of motor activity throughout the cycle and the after effects observed in the rhythm under DD depended on light intensity and access to the wheel. Moreover, under T23, some rats showed two simultaneous circadian components whose manifestation also depended on the experimental conditions. The results indicate that the strength of circadian entrainment to LD cycles in the rat depends on three factors: the period length of the LD cycle, light intensity used during the light phase, and access to a running wheel.

Constant Bright Light (LL) during Lactation in Rats Prevents Arhythmicity Due to LL

Light has a strong effect on the circadian system. Light-dark (LD) cycles are the main zeitgebers for practically all organisms, and the exposure of animals to constant bright light (LL) alters the manifestation of circadian rhythms. In rats, exposure to LL in adulthood produces an arrhythmic pattern in their motor activity, with a large number of ultradian components. In previous experiments, we found that rats born and kept under LL during lactation develop, after weaning, a circadian rhythm which is maintained for at least a couple of months. Here, we examined motor activity rhythms under LL of two groups of rats which differed in the lighting conditions under which they were kept during lactation: 1) rats kept under LL during lactation (LL-rats), which manifested a circadian rhythm after weaning, and 2) rats kept under constant darkness (DD-rats), which were arrhythmic after weaning. We investigated whether the presence of rhythmicity under LL in LL-rats is a transitory effect or whether it persists throughout most of the life of the rat. Moreover, we examined motor activity rhythms of both groups of rats under different lighting conditions to find out other possible differences in the manifestation of their circadian rhythms. Results showed that there are no differences in the capacity of entrainment of both groups of rats to LD cycles or in the rhythm that rats show under DD. Most of the LL-rats maintained their circadian rhythms for the duration of the experiment (1 year), although we found differences in the rhythms manifested between males and females. We found that most of the LL-males became arrhythmic; consequently, at the end of the experiment, there were no differences in the number of males showing circadian rhythm in the LL- and DD-groups. Most of the females in the LL-group showed a clear circadian rhythm under LL during the entire experiment. Thus, LL during lactation has a protective effect against the disruptive effect of LL on the circadian rhythm, although it is only clearly manifested in females.

Light responses of the circadian system in leptin deficient mice.

Some evidences postulate a link between obesity and disturbances in circadian behavior. Here, we studied the manifestation of the circadian rhythm of motor activity and its response to light in the leptin deficiency model of obesity ob/ob mice. Motor activity in both ob/ob and wild type mice was first recorded in a small cage by activity meters with crossed infrared beams (IR) and later in a larger cage with a running wheel, where the number of wheel revolutions (WR) was also determined. Animals were maintained under light-dark (LD) or constant-dark (DD) conditions. We studied the free-running period and the rhythm profile, with special emphasis on the amount of activity in the dark and light phases of the LD cycle, and the phase and period responses to a light pulse. The results showed that ob/ob mice have a strong ultradian, rather than a circadian pattern, whose period range between 3 and 4.8h. Also, these animals showed a percentage of activity during light higher than controls. We did not find differences in the endogenous period of the circadian rhythm between mice groups in DD. However, ob/ob mice showed stronger phase delays after a light pulse at ZT15 than controls, and less masking effects in the transition from LD to DD compared with controls. This suggests a weaker circadian pacemaker of the ob/ob mice compared with controls.

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.

Arrhythmic Rats after SCN Lesions and Constant Light Differ in Short Time Scale Regulation of Locomotor Activity

Circadian rhythm disruption (i.e., arrhythmicity) in motor activity is an abnormal behavioral pattern. In rats, it can be caused by the lesion of the hypothalamic suprachiasmatic nucleus (SCN) and by prolonged exposure to constant light (LL). We carried out a comparative study of these arrhythmic phenotypes to assess the role of the SCN in the regulation of the motor output beyond circadian rhythmicity. Motor activity series were studied in rats that had become arrhythmic as a result of 1) LL exposure at 2 light intensities: 300 lux (LL300) and 1.3 lux (LL1.3), and 2) SCN lesion (SCNx). The Fourier spectra, the fractal Hurst coefficient (H) from the autocorrelation function, and the β slope from the power spectral density were calculated in data sections at baseline, when the rats were still rhythmic, and later at stages with undetectable circadian rhythms. In the LL300 group, high power content was detected at frequencies of 8 to 4 h (i.e., ultradian). Lower power content for these harmonics was found in the LL1.3 group, whereas no dominant harmonics appeared in the SCNx group. Independently of the manifestation of circadian rhythm, H values were higher and more sustained in time in rats exposed to LL 300 but gradually decreased in rats exposed to LL1.3. Fractal correlation was found in control DD group but was absent in the SCNx group. We conclude that scale-invariant regulation of the motor pattern by SCN activity is dependent on light intensity but independent of the circadian rhythm output. Adjusting the light intensity by modifying the coupling degree between the population of oscillations could affect the dynamics of each individual oscillator in the SCN, making it less predictable.

Effect of light on the development of the circadian rhythm of motor activity in the mouse.

In previous experiments, we found that rats raised in constant light (LL) manifested a more robust circadian rhythm of motor activity in LL and showed longer phase shifts after a light pulse in constant darkness (DD) than those raised under constant darkness. In addition, we observed that the effects produced by constant light differed depending on the time of postnatal development in which it was given. These results suggest that both sensitivity to light and the functioning of the circadian pacemaker of the rat could be affected by the environmental conditions experienced during postembryonic development. Thus, the present experiment aimed to study whether postnatal exposure to light could also affect the circadian system of the mouse. Three groups of mice were formed: One group was raised under constant darkness during lactation (DD group), the second under constant light (LL group), and the third under light-dark cycles (LD group). After lactation, the three groups were submitted first to constant light of high intensity, then to LD cycles, and finally to constant darkness. In the DD stage, a light pulse was given. Finally, mice were submitted to constant light of low intensity. We observed that the circadian rhythm of the DD group was more disturbed under constant light than the rhythm of the LL group, and that, when light intensity increased, the period of the rhythm of the DD group lengthened more than that of the LL group. No significant differences among the groups were found in the phase shift induced by the light pulse. Therefore, it appears that DD mice are more sensitive to light than their LL counterparts. However, at present there is no evidence to affirm that the light environment experienced by the mouse during postnatal development affects the circadian pacemaker. (Chronobiology International, 18(4), 683–696, 2001)

Simultaneous Manifestation of Free-Running and Entrained Rhythms in the Rat Motor Activity Explained by A Multioscillatory System

Seven groups of 6 young male rats were exposed for 60 calendar days to symmetrical light/dark cycles with different periods (T = 22, 23, 24, 25, 26, 27, and 28 h), and subsequently to constant darkness for 30 days. During exposure to the light/dark cycles, all the animals in all groups presented a motor activity component that was entrained to the external cycles, although the animals subjected to light/dark cycles with periods shorter than 25 hours also presented a nonentrained circadian component. Moreover, in all cases, the effect of masking was present, manifested as a reactivity to the light-dark transitions and as a reduction of activity induced by light. Our results demonstrate that masking, entrainment, and free-running rhythm can be present at the same time. The simultaneous presence of the light-entrained component and the nonentrained component can be explained in terms of a multioscillatory system, in which some oscillators could be entrained to the light/dark cycles while other oscillators could be nonentrained at the same time. Thus, the present study indicates that the circadian system is not necessarily entirely entrained, and that the degree of entrainment may depend on the number of oscillators involved in generating the entrained rhythm.

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.