Roles of suprachiasmatic nuclei and intergeniculate leaflets in mediating the phase-shifting effects of a serotonergic agonist and their photic modulation during subjective day.

Serotonin (5-HT) has been implicated in the phase adjustment of the circadian system during the subjective day in response to nonphotic stimuli. Two components of the circadian system, the suprachiasmatic nucleus (SCN) (site of the circadian clock) and the intergeniculate leaflet (IGL), receive serotonergic projections from the median raphe nucleus and the dorsal raphe nucleus, respectively. Experiment 1, performed in golden hamsters housed in constant darkness, compared the effects of bilateral microinjections of the 5-HT1A/7 receptor agonist, 8-hydroxydipropylaminotetralin (8-OH-DPAT; 0.5 microgram in 0.2 microliter saline per side), into the IGL or the SCN during the mid-subjective day. Bilateral 8-OH-DPAT injections into either the SCN or the IGL led to significant phase advances of the circadian rhythm of wheel-running activity (p < .001). The phase advances following 8-OH-DPAT injections in the IGL were dose department (p < .001). Because a light pulse administered during the middle of the subjective day can attenuate the phase-resetting effect of a systemic injection of 8-OH-DPAT, Experiment 2 was designed to determine whether light could modulate 5-HT agonist activity at the level of the SCN and/or the IGL. Serotonergic receptor activation within the SCN, followed by a pulse of light (300 lux of white light lasting 30 min), still induced phase advances. In contrast, the effect of serotonergic stimulation within the IGL was blocked by a light pulse. These results indicate that the respective 5-HT projections to the SCN and IGL subserve different functions in the circadian responses to photic and nonphotic stimuli.

Daily social and physical activity increases slow-wave sleep and daytime neuropsychological performance in the elderly.

Decreased levels of physical and social activity associated with aging can be particularly pronounced in residents of assisted living facilities. Reduced exposure to important behavioral and time-giving cues may contribute to the age-related changes in circadian rhythmicity and sleep. The present study was conducted to test the hypothesis that an enforced schedule of structured social and physical activity (0:900 to 10:30 and 19:00 to 20:30 daily for two weeks) can have beneficial effects on circadian rhythmicity, nocturnal sleep, daytime functioning, mood, and vigor. The subjects were 14 elderly residents of continued-care retirement facilities while a similar group of 9 elderly residents served as controls. The group exposed to structured activities had increased amounts of slow-wave sleep and demonstrated improvement in memory-oriented tasks following the intervention. Conversely, no significant changes were noted in the amplitude and phase of the body temperature rhythm or in subjective measures of vigor and mood. These results indicate that short-term exposure to structured social intervention and light physical activity can significantly improve memory performance and enhance slow-wave sleep in older adults without alterations to the circadian phase or amplitude of body temperature. This is the first report to demonstrate that low intensity activity in an elderly population can increase deep sleep and improve memory functioning. The high degree of interest in these activities paired with the simple nature of the tasks makes this a potentially practical intervention which can be adapted for both community dwelling and assisted-living elders.

A serotonin neurotoxin attenuates the phase-shifting effects of triazolam on the circadian clock in hamsters.

Several lines of evidence suggest the potential involvement of serotonergic pathways in mediating the effects of activity-inducing stimuli on the circadian clock in rodents. The aim of the present 3 experiments was to examine the effects of the serotonergic neurotoxin, p-chloroamphetamine (PCA, 10 mg/kg) on: (1) the monoamine levels of the hypothalamus, frontal cortex and hippocampus in the hamster; (2) the phase shifts in the circadian rhythm of locomotor activity of hamsters in response to treatment with the short-acting benzodiazepine, triazolam (7.5 mg/kg); and (3) the magnitude of the acute increase in locomotor activity associated with triazolam administration in this species. The administration of PCA to hamsters caused changes of specific monoaminergic systems in the hypothalamus, that were limited to a selective decrease in serotonin levels 7 days post-treatment. The phase shifts of the circadian clock in response to triazolam treatment at CT 6 were considerably attenuated following the administration of the 5-HT neurotoxin. The total amount and the profiles of triazolam-induced wheel-running and general cage activity between CT 6 and CT 12 were not significantly affected by the PCA treatment. The finding that a 5-HT neurotoxin can attenuate the phase-shifting effects of triazolam in hamsters, without interfering with its activity-inducing properties, suggests that serotonergic afferents might be involved in the mechanism for non-photic phase-shifting of the circadian system.

Monoamine depletion blocks triazolam-induced phase advances of the circadian clock in hamsters.

Injections with the short-acting benzodiazepine, triazolam (Tz), 6 h before activity onset (CT6) produce large phase advances of the circadian pacemaker in hamsters. An increase in locomotor activity and/or the state of arousal is considered essential for the effects of Tz, suggesting the potential involvement of central monoaminergic systems in this process. The present study examines the effect of reserpine-induced monoamine depletion on the phase-shifting effects of Tz in hamsters. Wheel running activity of 16 male golden hamsters (14 weeks old) was continuously monitored in constant darkness. After a stable free-running circadian rhythm was established half of the animals received reserpine (2.5 mg/kg, s.c.) and the other half vehicle treatment. Ten days later all animals were given Tz injections (10 mg/kg i.p.) at CT6 and the circadian activity rhythm was monitored for 2 more weeks. An additional 10 animals were used to determine the effect of reserpine on the central monamine levels using high pressure liquid chromatography. A circadian rhythm of locomotor activity with reduced amplitude and longer free-running period persisted after reserpine treatment, despite the significant monoamine depletion. Triazolam injections at CT6 induced large phase-advances (93.1 +/- 14.9) in the control group that were markedly attenuated in 7 out of the 8 reserpine-treated animals (3.12 +/- 17.7 min, P < 0.01). Our results suggest that monoaminergic systems are essential for the phase-shifting effect of Tz upon the circadian system in hamsters.

Monoamine depletion alters the entrainment and the response to light of the circadian activity rhythm in hamsters.

Reduced amplitude, shorter free-running periods and desynchronization among a number of circadian rhythms are associated with advanced age in rodents. The response of the hamster circadian system to photic stimuli is also altered during senescence. Decreased monoamine levels, receptor sites and neuronal populations are commonly observed in the aging brain. The objective of the present study was to determine if monoamine depletion with reserpine in young hamsters induces changes in the circadian rhythm of locomotor activity similar to those that occur spontaneously with aging. Wheel-running activity of 12 young hamsters under a 14 h-light/10 h-dark cycle was continuously monitored. The total activity level, the times of activity onset, peak and offset and the duration of activity were determined during a 1-week period after vehicle treatment and for three 1-week periods after reserpine treatment (4 mg/kg). A second group of eight reserpine-treated and six vehicle-treated animals was kept in constant darkness (DD). The period of the circadian activity rhythm in DD and the phase-shifts after short light pulses at circadian time 19 (CT19) were determined in the control and reserpine-treated groups. Brain monoamines in the hypothalamus, striatum and pons/medulla after reserpine and vehicle treatment were determined by high-pressure liquid chromatography. The data were analyzed with x2 periodogram and one-way ANOVA followed by Duncan's post hoc test. Reserpine treatment significantly reduced total wheel-running activity and the monoamine levels in the hypothalamus, striatum and pons/medulla.(ABSTRACT TRUNCATED AT 250 WORDS)

Phase-shifting effects of a serotonin agonist in tau mutant hamsters.

Previous studies indicate that the advance region of the tau mutant hamster's phase-response curve (PRC) to non-photic stimuli, such as NPY and wheel pulses, is characterized by earlier timing and increased amplitude in comparison with that of wild-type animals. Since, recent evidence suggests that serotonergic pathways may play an important role for the non-photic phase resetting of the rodent circadian pacemaker, PRCs to the serotonin (5-HT) agonist, 8-OH-DPAT (5 mg/kg i.p), were generated in both wild-type and tau mutant hamsters kept in constant darkness. The results indicate that the tau mutation is associated with changes in the timing, but not the amplitude of the advance region of the PRC to 8-OH-DPAT and suggest that serotonergic agents and other non-photic or activity-inducing stimuli may share some common mechanisms for resetting the phase of the rodent circadian pacemaker.

Blood pressure and heart rate rhythmicity: differential effects of late pregnancy.

Arterial blood pressure (BP) and heart rate (HR) of 31 hospitalized pregnant women at low risk of hypertension were automatically monitored for 48 h at 15-min intervals. Each of the recorded 56 data series for systolic arterial pressure (SAP), diastolic arterial pressure (DAP), and HR was chronobiologically assessed by linear-nonlinear rhythmometry. The rhythm-adjusted mean (MESOR), circadian amplitude, circadian acrophase, and best-fitting period were grouped by pregnancy trimester and further subjected to analysis of variance. BP MESOR remained unaltered, whereas HR MESOR increased significantly in middle and late pregnancy. Ultradian rhythms, with an amplitude higher than that of the circadian rhythm, were found in 25% of the SAP records in the second and third trimester. Such ultradian rhythms were not detected in the simultaneously recorded HR. Finally, the group BP and HR circadian acrophases coincided in the first trimester, but were significantly apart in mid and late pregnancy. These observations support the notion that the coordination of BP and HR rhythmicity involves different physiological mechanisms. Analysis of the individual variability in the chronobiological end points (based on the records of nine women monitored in each pregnancy trimester) revealed that only the BP MESOR was well reproducible in the course of pregnancy and may be useful in early diagnosis of gestational hypertension.

Suprachiasmatic nuclei lesions eliminate light/dark variations in short-term feeding responses to deprivation or insulin treatment in rats.

Short-term (1-h, 4-h and 12-h) and long-term (24-h) feeding responses to 24-h food deprivation (FD) or insulin treatment (IT) (8 mU/kg IP) were studied in male rats under a 12/12-h light/dark (L/D) cycle. The 24-h FD or the IT began either at onset (Dawn) or offset (Dusk) of the lights. In sham-operated rats (Shams) both protocols elicited greater short-term feeding responses at Dusk (p less than 0.05 or less). In suprachiasmatic nuclei (SCN)-lesioned rats the L/D variations in short-term responses were absent. In both SCN and Shams the 24-h feeding responses did not depend on stimuli time-schedule. We conclude that the regulation of short-term (circadian), but not long-term, feeding responses to metabolic stimuli is dependent on SCN integrity in the rat.

[Dynamics of agglutinins following vaccination against coli-bacteriosis in calves]. / Dinamika na aglutininite pri vaktsinatsiia protiv kolibakterioza po teletata.

A bivalent vaccine, produced with strains Escherichia coli 078 and 0117 to control colibacteriosis in calves, was used to vaccinate experimentally 20 cows following up the agglutinin dynamics in the serum of calves born later on. It was found that the agglutinin titers of the 0 antibodies were lower than those of OK, and varied within the range of from 1:10 to 1:320, the titers of 1:20-1:40 values being prevalent. Highest level of titers was observed by the third day following birth. OK antibodies had titers that were 1-2 degrees higher, reaching their peak level as early as the 12-15 hour after birth. They persisted for a longer period in the blood serum of calves. The antibodies produced after vaccination were shown to be sensitive to 2-mercaptoethanol.

Melatonin: a clinical perspective.

Over the last several decades the pineal gland has emerged as an active neuroendocrine transducer of important environmental information. However, the current understanding of the function of its major hormone, melatonin, in humans remains ill defined and based exclusively on correlative observations. In a similar manner, the multitude of phenomenological descriptions of the effects of exogenous melatonin is contrasted by the limited understanding of the underlying mechanisms and the lack of firmly established clinical applications for the hormone. Future randomized, double-blind, placebo-controlled clinical studies will be necessary to determine the precise indications, treatment regimens, and safety of melatonin in clinical practice. The recent rapid progress in the area of melatonin research should lead to a better understanding of its role in human health and disease.

Quantitative analysis of the age-related fragmentation of hamster 24-h activity rhythms.

The continuous monitoring of spontaneous locomotor activity has emerged as one of the most widely used metrics in rodent circadian research. This behavioral measure is also extremely useful for the description of the effects of aging on circadian rhythms. The present study describes the successful use of a log-survivorship approach to identify discrete bouts of hamster wheel-running activity and provides a detailed description of the age-related fragmentation in the 24-h profile of this behavioral variable. In addition, stepwise discriminant analysis identified the most important quantitative measures for distinguishing between the individual patterns of wheel-running activity of young (3 mo) and old (17-18 mo) golden hamsters. The results suggest that this method of bout analysis can be a valuable tool for the study of genetic, developmental, neurochemical, physiological, and environmental factors involved in the temporal control of rodent locomotor behavior.

A reliable technique for midline stereotaxic cannulation of the third ventricle in the rat.

A simple technique is described for chronic or acute cannulation of the third ventricle in the rat slightly displacing the superior sagittal sinus to allow midline implantation of a vertically oriented cannula. The results of an experiment successfully employing the technique for transplantation of embryonic hypothalamic tissue in rats are briefly described.

Aging alters the serotonergic modulation of light-induced phase advances in golden hamsters.

Recent findings have raised the possibility that some of the age-related changes in the circadian system and the response of the circadian pacemaker to environmental stimuli may involve central serotonergic mechanisms. The present study compared the effects ofpretreatment with the serotonin agonist 8-hydroxy-2(di-n-propylamino)tetralin (5 mg/kg ip) on the magnitude of light-induced phase advances in young (2-4 mo) and old (18-20 mo) golden hamsters. The ability of this serotonin agonist to attenuate the photic phase resetting of circadian locomotor rhythmicity in young animals was decreased by 46% in old hamsters (P < 0.05). These results suggest that deficits in the mechanisms for serotonergic control of circadian function may interfere with the optimal adaptation of the senescent organism to its temporal environment.

Effect of transplantation of embryonic anterior hypothalamic tissue from spontaneously hypertensive rats to normotensive Wistar rats on circadian rhythms of systolic arterial pressure and heart rate.

The anterior hypothalamus (AH) participates in the regulation of arterial pressure. The suprachiasmatic nuclei (SCN) of the AH are a major circadian oscillator necessary for the generation and/or the entrainment of circadian rhythms. Circadian rhythms of systolic arterial pressure (SAP) and heart rate (HR) were investigated in spontaneously hypertensive rats (SHR) and in normotensive Wistar rats (NWI) with intact SCN, grafted with SHR embryonic AH tissue containing the SCN. Prominent circadian rhythms for SAP and HR in both NWI and SHR with acrophases during dark were found. The elevation of the MESOR (midline-estimated statistic of rhythm) of the SAP in normotensive rats grafted with AH embryonic tissue obtained from SHR was accompanied by disappearance of the circadian rhythm of SAP. This result suggests an interaction between the grafted tissue containing the SCN on the one hand, and the host SCN on the other hand. Our data ascribe a role for the SCN in the entrainment of the circadian rhythm of arterial pressure. The circadian rhythm of HR was not eliminated by the SCN graft in spite of the amplitude decrease and the phase delay observed. It seems that the entrainment of the circadian rhythm of HR is probably not crucially dependent on the SCN in rats. The circadian rhythms of SAP and HR in rats were differently affected by the grafts, thus suggesting a multioscillatory system for circadian regulation in rats.

Chronic circadian desynchronization decreases the survival of animals with cardiomyopathic heart disease.

Shift work is associated with increased cardiovascular morbidity and mortality. Whereas it has been suggested that continuous shifting of the circadian clock/sleep-wake cycle may have negative effects on health, there is very little experimental evidence to support such a hypothesis. Cardiomyopathic Syrian hamsters were either maintained on a fixed light-dark (LD) cycle (n = 31) or were subjected to a 12-h phase shift in the LD cycle on a weekly basis (n = 32). The duration of the life span was recorded for each animal. Chronic reversal of the external LD cycle at weekly intervals resulted in a significant decrease in the survival time in cardiomyopathic hamsters with the median life span being reduced by 11%. Disrupting normal circadian rhythmicity in an animal susceptible to early mortality due to cardiac disease results in a further decrease in longevity. The deleterious effects of the chronic phase shifts in the LD cycle in cardiomyopathic hamsters may be related to reports of increased cardiovascular morbidity and mortality in humans engaged in shift work.

Suprachiasmatic nuclei lesions eliminate the group circadian rhythm of systolic arterial pressure but not of heart rate in rats.

The aim of this study was to evaluate the participation of the suprachiasmatic nuclei (SCN) in the generation and synchronization of cardiovascular rhythms. Seven sham-operated and 11 SCN-lesioned animals maintained under 12/12 hr light/dark cycle were used. Systolic arterial pressure (SAP) and heart rate (HR) were measured indirectly during 24-hour periods at 3-4 hour intervals. The data were analyzed using individual and group cosinor rhythmometry and Fourier analysis. A circadian rhythm of water intake was not detected in animals with successful SCN lesions. A reduction of the double amplitude/MESOR ratio for the 24-hour component of drinking rhythm in the SCN-lesioned rats was observed. After SCN lesions the group 24-hour rhythm of SAP was eliminated while a significant group circadian rhythm for HR was detected. The individual amplitude/MESOR ratios for the 24-, 12-, 8- and 6-hour periodic components of SAP and HR in the lesioned rats showed no marked differences as compared with controls. The generation and entrainment of circadian variations in HR is probably not dependent on the integrity of SCN in rats. The SCN may participate in the entrainment of the circadian rhythm of SAP. The combination of completely abolished (water intake) and persisting (heart rate) rhythms further supports that the circadian regulatory system consists of a network of multiple oscillators.

Aging alters the phase-resetting properties of a serotonin agonist on hamster circadian rhythmicity.

Serotonergic mechanisms are believed to play a considerable role in mediating the effects of photic and nonphotic stimuli on circadian rhythmicity. Because aging is associated with significant changes in the responsiveness of the rodent circadian system to major synchronizing agents in the environment, this study examined the phase-shifting effects of the 5-HT1A receptor agonist, 8-hydroxy-2-(di-n-propylamino)tetralin [8-OH-DPAT; 2.0, 5.0, and 8.0 mg/kg ip at circadian time 8 (CT 8)], on the 24-h activity rhythm of young (3-4 mo old) and old (18-19 mo old) golden hamsters. Aging was associated with a dramatic attenuation of the phase-shifting effects of 8-OH-DPAT in this species. The results suggest the existence of age-related deficits in the serotonergic control of mammalian circadian rhythmicity, which could interfere with the temporal adaptation of the senescent organism to its environment.

Pharmacological and genetic approaches for the study of circadian rhythms in mammals.

Two different approaches have been utilized to study the controlling mechanisms that underlie the generation and entrainment of circadian rhythms in mammals. The use of specific drugs to alter the period and/or the phase of circadian rhythms has provided new insights into both the pathways by which environmental information reaches the mammalian circadian pacemaker in the suprachiasmatic nuclei (SCN) and the cellular and neurochemical events within the SCN itself which are involved in circadian rhythmicity. A second approach, which seeks to exploit genetic differences in the properties of the circadian system, holds the promise of eventually defining the cellular and molecular events that are part of the clock itself, the events that underlie the entrainment of the circadian clock by environmental factors, and the expression of overt rhythms driven by the clock. It is anticipated that the pharmacological and genetic approaches to the study of circadian rhythms will complement each other as the underlying physiological mechanisms of the circadian clock system become defined.