Olivier Van Reeth

Effects of age on the circadian system

While aging has been associated with changes in the period and amplitude of circadian rhythms, little is known about how aging influences the response of the circadian clock to environmental stimuli. In this paper, we report on recent studies designed to determine the effects of advanced age on the response of the circadian clock to both photic and nonphotic stimuli in old hamsters (e.g., over 16 mo of age). Among the most pronounced age-related changes in the circadian rhythm of locomotor activity are: (a) alterations in the phase-angle of entrainment to the light-dark cycle; (b) an increase in the magnitude of phase shifts induced by pulses of light presented at specific circadian times; and (c) a loss of responsiveness to the phase shifting or entraining effects of stimuli which induce an acute increase of activity. Depletion of brain monoamine levels in young animals can induce changes in the responsiveness of the circadian clock to environmental stimuli which are similar to those which occur spontaneously in old animals, suggesting that aging alters monoaminergic inputs to the clock. Some of the age-related changes in the response of the clock to an activity-inducing stimulus can be reversed by implanting old animals with fetal SCN tissue. Determining the physiological basis for age related changes in the responsiveness of the clock to both internal and external stimuli, and the mechanisms by which normal circadian function can be restored, should lead to new insight into the functioning of the circadian clock and may lead to new approaches for normalizing disturbed circadian rhythms.

Sleepiness, performance, and neuroendocrine function during sleep deprivation: effects of exposure to bright light or exercise.

The temporal profiles of subjective fatigue (as assessed by the Stanford Sleepiness Scale), of cognitive performance (on a digit symbol substitution test and a symbol copying task), of body temperature, and of the peripheral concen trations of melatonin, thyroid-stimulating hormone (TSH), and cortisol were obtained simultaneously at frequent intervals in 17 normal young subjects submitted to a 43-h period of constant routine conditions involving continuous wakefulness at bed rest in dim indoor light. The subjects had knowledge of time of day. Caloric intake was exclusively in the form of an intravenous glucose infusion, and plasma glucose levels were monitored continuously in 8 of the 17 subjects. Under these conditions, fluctuations in plasma glucose reflect primarily changes in glucose use because endogenous glucose production is suppressed by the exogenous infusion. Following the completion of a baseline constant routine study, the volunteers participated in two subsequent studies using the same protocol to determine the immediate psychophysiological effects of expo sure to a 3-h pulse of bright light or to a 3-h pulse of physical exercise. Sleepiness and performance varied in a mirror image, with significant negative correlations. Sleepiness scores were minimal around noon and then increased at a modest rate throughout the rest of the normal waking period. Staying awake during usual bedtime hours was associated with an acceleration in the rate of increase in sleepiness, which coincided with decreasing body temperature, rapidly rising cortisol concentrations, and maximal levels of melatonin and TSH. When body temperature reached its nadir, a further major increase in sleepiness occurred in parallel with a pronounced decrease in plasma glucose (reflecting increased glucose use). Recovery from maximal sleepiness started when blood glucose levels stopped falling and when significant decreases in cortisol and melatonin concentrations were initiated. Lower levels of subjective sleepiness resumed when glucose concentrations and body temperature had returned to levels similar to those observed prior to sleep deprivation and when melatonin and TSH concentrations had returned to daytime levels. The synchrony of behav ioral, neuroendocrine, and metabolic changes suggests that circulating hormonal levels could exert modulatory influences on sleepiness and that metabolic altera-tions may underlie the sudden increase in fatigue consistently occurring at the end of a night of sleep deprivation. Effects of bright light or exercise exposure on subjective sleepiness appeared to be critically dependent on the timing of exposure.

Sleep deprivation decreases phase-shift responses of circadian rhythms to light in the mouse: role of serotonergic and metabolic signals

The circadian pacemaker in the suprachiasmatic nuclei is primarily synchronized to the daily light-dark cycle. The phase-shifting and synchronizing effects of light can be modulated by non-photic factors, such as behavioral, metabolic or serotonergic cues. The present experiments examine the effects of sleep deprivation on the response of the circadian pacemaker to light and test the possible involvement of serotonergic and/or metabolic cues in mediating the effects of sleep deprivation. Photic phase-shifting of the locomotor activity rhythm was analyzed in mice transferred from a light-dark cycle to constant darkness, and sleep-deprived for 8 h from Zeitgeber Time 6 to Zeitgeber Time 14. Phase-delays in response to a 10-min light pulse at Zeitgeber Time 14 were reduced by 30% in sleep-deprived mice compared to control mice, while sleep deprivation without light exposure induced no significant phase-shifts. Stimulation of serotonin neurotransmission by fluoxetine (10 mg/kg), a serotonin reuptake inhibitor that decreases light-induced phase-delays in non-deprived mice, did not further reduce light-induced phase-delays in sleep-deprived mice. Impairment of serotonin neurotransmission with p-chloroamphetamine (three injections of 10 mg/kg), which did not increase light-induced phase-delays in non-deprived mice significantly, partially normalized light-induced phase-delays in sleep-deprived mice. Injections of glucose increased light-induced phase-delays in control and sleep-deprived mice. Chemical damage of the ventromedial hypothalamus by gold-thioglucose (600 mg/kg) prevented the reduction of light-induced phase-delays in sleep-deprived mice, without altering phase-delays in control mice. Taken together, the present results indicate that sleep deprivation can reduce the light-induced phase-shifts of the mouse suprachiasmatic pacemaker, due to serotonergic and metabolic changes associated with the loss of sleep.

Fos protein expression in the circadian clock is not associated with phase shifts induced by a nonphotic stimulus, triazolam

Recent studies have shown that light-induced phase shifts of the circadian rhythm of locomotor activity are associated with c-fos expression in the suprachiasmatic nucleus (SCN) and intergeniculate leaflet (IGL) of the lateral geniculate nucleus of rodents. In order to determine whether c-fos expression is necessary for the phase shifting effects of a non-photic stimulus, we assessed Fos-like immunoreactivity (Fos-lir) in the SCN and IGL at various times after an injection of the short-acting benzodiazepine, triazolam, at circadian time (CT) 6; i.e. at a time when triazolam induces an acute increase in locomotor activity and maximal phase advances in the circadian rhythm of locomotor activity. Specific Fos-lir staining was not observed in the SCN or IGL regions of any animals treated with triazolam or vehicle at any time point examined. These results indicate that exposure to an activity-inducing stimulus at circadian times when this stimulus induces phase shifts does not induce Fos protein synthesis in the SCN or IGL regions.

Resynchronisation of a diurnal rodent circadian clock accelerated by a melatonin agonist.

USING ‘jet lag’ paradigms involving phase shifts in the light-dark (LD) cycle, we studied the effects of S-20098 on the circadian clock of a diurnal rodent. Arvicanthis mordax, entrained to a regular LD cycle, were subjected to advance shifts (i.e. 4, 6 or 8 h) in the LD cycle and injected with vehicle or the melatonin agonist S-20098 (20 mg/kg) the day of the shift (and also on subsequent days in the 6 h or 8 h shift paradigms). In each condition, S-20098 accelerated by about 30% resynchronization to the new LD cycle. These data, which are the first to demonstrate the chronobiotic effects of a melatonin agonist in a diurnal rodent, provide new insights for the design of human chronopharmacological protocols.

Daily variations of blood glucose, acid-base state and PCO2 in rats: effect of light exposure.

The suprachiasmatic nuclei (SCN) of the hypothalamus are the site of the main circadian clock in mammals. Synchronization of the SCN to light is achieved by direct retinal inputs. The present study performed in rats transferred to constant darkness shows that blood glucose, pH and PCO2 display significant diurnal changes when measurements were made during the subjective day, the early subjective night or the late subjective night. The effects of a 30-min light exposure (100 lx) on these metabolic parameters at each of these circadian times were assessed. Regardless of the circadian time, light induced an increase in blood glucose, but did not affect plasma pH and PCO2. This study suggests that blood glucose, PCO2 and acid-base state are under circadian control, most likely mediated by the SCN, while the hyperglycemic response to light seems not to be gated by a circadian clock and may thus involve retinal inputs to non-SCN retino-recipient areas.

Sleep and Circadian Disturbances in Shift Work: Strategies for Their Management

More and more businesses are providing their full range of services 24 h a day, 7 days a week, thus forcing their employees to work either rotating shifts or fixed night shifts. Dictates of our endogenous circadian clock prevent our brains and bodies to be indefinitely adaptable to those work schedules. Shift work operations are thus associated with serious healthy and social problems for the workers. Various interventions can counteract circadian desynchronization, sleep disturbances, and social disruption associated with shift work: changes in work schedules, sleeping and napping strategies, use of appropriately timed exposure to bright light, experimental drug treatments, or exercise.

Phase shifts in the circadian activity rhythm induced by triazolam are not mediated by the eyes or the pineal gland in the hamster

A single injection of the benzodiazepine triazolam, which is though to act by potentiating the effects of the neurotransmitter gamma-aminobutyric acid (GABA), can induce permanent phase shifts in the circadian rhythm of locomotor activity of hamsters. Occurrence of GABA immunoreactivity and benzodiazepine receptors in the retina, which contains photoreceptors that relay synchronizing light-dark information to the mammalian circadian system, raises the possibility that triazolam may influence circadian rhythmicity via an action on the retina. However, the phase shifting effects of triazolam on the activity rhythm were unaffected by blinding: the direction and the magnitude of the phase shifts were similar in blind hamsters and in sighted hamsters maintained in constant darkness. Furthermore, no change in response to triazolam was observed in hamsters studied through 84 days after blinding. In addition, benzodiazepine binding sites have been found in the mammalian pineal gland, which has also been implicated in circadian rhythmicity. Therefore, its possible involvement in mediating the phase advancing effects of triazolam on the circadian clock has also been tested: the response was similar in blind and blind-pinealectomized animals. These results indicate that the effects of triazolam on the circadian clock are not mediated by the eyes or the pineal gland.

Altered circadian responses to light in streptozotocin-induced diabetic mice

Diabetes mellitus affects the daily expression of many behavioral and metabolic processes. Recent studies indicate that changes in brain glucose metabolism alter the entraining effects of light of the circadian pacemaker. To test whether diabetes-associated diurnal changes are related to alterations in the responses of the circadian pacemaker to light, photic phase resetting of the circadian rhythm of locomotor activity was analyzed in diabetic mice housed in constant darkness. Multiple low doses of streptozotocin, which damages pancreatic beta-insulin-producing cells, were used to render C57BL/6J mice mildly diabetic. In those mice treated with streptozotocin, serum glucose was increased by 25% and circadian responses to light either were increased by 40% for phase delays or were close to those observed in control animals for phase advances. Furthermore, insulin-induced hypoglycemia normalized light-induced phase delays in diabetic animals, without altering those in nondiabetic mice. These results show that abnormalities of daily temporal organization associated with diabetes can result from altered circadian responses to the daily variation in ambient light. Such alterations could be normalized with appropriate insulin therapy.Diabetes mellitus affects the daily expression of many behavioral and metabolic processes. Recent studies indicate that changes in brain glucose metabolism alter the entraining effects of light of the circadian pacemaker. To test whether diabetes-associated diurnal changes are related to alterations in the responses of the circadian pacemaker to light, photic phase resetting of the circadian rhythm of locomotor activity was analyzed in diabetic mice housed in constant darkness. Multiple low doses of streptozotocin, which damages pancreatic β-insulin-producing cells, were used to render C57BL/6J mice mildly diabetic. In those mice treated with streptozotocin, serum glucose was increased by 25% and circadian responses to light either were increased by 40% for phase delays or were close to those observed in control animals for phase advances. Furthermore, insulin-induced hypoglycemia normalized light-induced phase delays in diabetic animals, without altering those in nondiabetic mice. These results show that abnormalities of daily temporal organization associated with diabetes can result from altered circadian responses to the daily variation in ambient light. Such alterations could be normalized with appropriate insulin therapy.

Autoradiographic localization of cholecystokinin binding sites in human cerebellar system using a [125I]CCK8 probe

Carboxyl-terminal cholecystokinin octapeptide (CCK8) binding sites were studied in the human cerebellar system by autoradiography. High affinity CCK8 binding sites were demonstrated in the main cerebellar afferent nuclei, namely the inferior olivary complex and the pontine nuclei. This localization of CCK8 binding sites was partly correlated with already described CCK containing terminals. In the cerebellar cortex, high affinity CCK8 binding sites were detected with a laminar distribution. Levels were higher in the granular layer (mostly in the superficial part) and lower in the white matter and the Purkinje cell layer. The non-specific binding was homogenous and particularly low (9%) in the cerebellar cortex but a non-specific binding was selectively localized in the deep cerebellar nuclei. Those results illustrate the species variability of CCK binding sites in the cerebellum and are briefly discussed in relation with the low level of CCK immunoreactivity in this structure. The presence of CCK8 binding sites in cerebellar afferent nuclei and cortex suggests a role of CCK in human cerebellar physiology and particularly in the modulation of afferent inputs to the cerebellum.