Darkness during early postnatal development is required for normal circadian patterns in the adult rat.

Early light experience influences the brain during development. Perinatal light exposure has an important effect on the development of the circadian system, although the role of quantity versus quality of light in this process is still unclear. We tested the development of the circadian rhythm of locomotor activity under constant bright light from the day of weaning, of six groups of rats raised under different light conditions during suckling. Results indicated that when rats received daily darkness during suckling (rats reared under constant darkness or light-dark cycles with dim or bright light) became arrhythmic when exposed to continuous bright light after weaning. However, those rats reared in the absence of darkness (constant dim or bright light, or alternating dim and bright light) developed a circadian rhythm, which was stronger and had a shorter period depending on the quantity of light received during suckling. Vasointestinal polypeptide immunoreactivity in the suprachiasmatic nucleus (SCN) was higher in those rats with weaker rhythms. However, no apparent differences among these groups were found in the melanopsin-expressing retinal ganglion cells, which provide the SCN with light input in the photoentrainment process. When bright light was shifted to dim light in three of the groups on day 57 after weaning, all of them generated a circadian rhythm with a longer period in those rats previously arrhythmic. Our results indicate the importance of the amount of light received at the early stages of life in the development of the circadian system and suggest that darkness is needed for the normal development of circadian behaviour.

Constant bright light (LL) during lactation in rats prevents arrhythmicity 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.

Period length of the light-dark cycle influences the growth rate and food intake in mice.

The adaptation of the endogenous rhythm of an organism to external cycles may be critical for the development of physiological processes in which energy is expended. We sought to determine whether growth rate depends on the degree of tuning between the external cycle and the manifestation of the circadian rhythms. To do so, we studied the growth rate and the food intake of mice (seven groups of 20 animals each) kept under symmetric LD cycles with different periods (T) of 21, 22, 23, 24, 25, 26, and 27 h, respectively, for 80 days. The mice were then kept in constant darkness for a further 80 days. Throughout the experiment, motor activity was recorded every 15 min for each mouse by means of an actimeter that used crossed infrared beams. Several variables related to the circadian motor activity rhythm were calculated, and correlated with body weight, food intake, and the efficiency of food for growth (food efficiency) calculated as: 100 x body weight increase/ amount of food intake. Results show that these three variables seem to be influenced by the number of circadian cycles that the animal has experienced, but also, and more significantly, by the degree to which the alpha phase of the individual rhythm and the dark phase of the external cycle coincide. Therefore, circadian rhythms would seem to affect the physiological processes that regulate growth and energy expenditure.

Expression of circadian neuropeptides in the hypothalamus of adult mice is affected by postnatal light experience.

The suprachiasmatic nuclei (SCN) of the hypothalamus are the principal pacemaker in mammals, controlling daily, circadian rhythms in physiology and behaviour. Environmental light during development has long-term effects on circadian behaviour, but it is still unclear what the relevant adaptations within the brain are. In the present study, we examined the manifestation of the circadian rhythm of locomotor activity, and the expression of arginine-vasopressin (AVP) and vasointestinal polypeptide (VIP) in the SCN of adult mice reared under different light environments during the suckling period, and synchronised to light/dark cycles after weaning. We found that animals reared under constant light had higher amplitude and more stable activity rhythms, together with lower levels of VIP- and AVP-immunostaining in the SCN, compared to mice reared under light/dark cycles or constant darkness. Differences in AVP expression were also found in the paraventricular nucleus and the supraoptic nucleus, two brain areas which receive SCN projections. These results indicate that the postnatal light experience may affect clock function and clock output, and suggest implications for the control of hormonal homeostasis and circadian behaviour.

Resetting of the hamster circadian system by dark pulses.

Circadian rhythms of animals are reset by exposure to light as well as dark; however, although the parameters of photic entrainment are well characterized, the phase-shifting actions of dark pulses are poorly understood. Here, we determined the tonic and phasic effects of short (0.25 h), moderate (3 h), and long (6-9 h) duration dark pulses on the wheel-running rhythms of hamsters in constant light. Moderate- and long-duration dark pulses phase dependently reset behavioral rhythms, and the magnitude of these phase shifts increased as a function of the duration of the dark pulse. In contrast, the 0.25-h dark pulses failed to evoke consistent effects at any circadian phase tested. Interestingly, moderate- and long-dark pulses elevated locomotor activity (wheel-running) on the day of treatment. This induced wheel-running was highly correlated with phase shift magnitude when the pulse was given during the subjective day. This, together with the finding that animals pulsed during the subjective day are behaviorally active throughout the pulse, suggests that both locomotor activity and behavioral activation play an important role in the phase-resetting actions of dark pulses. We also found that the robustness of the wheel-running rhythm was weakened, and the amount of wheel-running decreased on the days after exposure to dark pulses; these effects were dependent on pulse duration. In summary, similarly to light, the resetting actions of dark pulses are dependent on both circadian phase and stimulus duration. However, dark pulses appear more complex stimuli, with both photic and nonphotic resetting properties.

Manifestation of circadian rhythm under constant light depends on lighting conditions during lactation.

Adult rats transferred to continuous illumination (LL) show a disruption of circadian rhythms, although the mechanisms underlying this effect are not yet well known. In previous experiments, we found that when rats were born and raised under LL they showed an ultradian pattern during the first 10 days after weaning, but afterward they generated a circadian rhythm that was maintained until adulthood. It was not clear whether this evolution was attributable to the influence of the rhythm of the mother or to the effect of constant light. Here, we have studied the motor activity rhythm of young rats maintained under LL after weaning, taking into account the conditions to which they were exposed during lactation [LL or continuous darkness (DD)]. To check the possible effect of the rhythm of the dam, on the day of delivery some of the dams were blinded, others were subjected to a restricted feeding schedule of 3 h/day, and the others were used as controls. For each rat, the period of the circadian rhythm and the percentage of variance explained by this rhythm were calculated. Results show that all rats maintained under LL during lactation expressed a circadian rhythm in their motor activity. However, rats maintained under DD during lactation did not. This effect did not seem to be dependent on the type of dam. These results suggest that the rhythm of the dams does not affect the manifestation of the rhythm of the pups and that the expression of circadian rhythmicity under constant bright light depends on the lighting conditions under which the animals were maintained during lactation, which could affect the development of the circadian pacemaker or the retina.