Tobacco exposure and sleep disturbance in 498 208 UK Biobank participants.

Background: The prevalence of sleep disturbance is high and increasing. The study investigated whether active, former and passive smoking were associated with sleep disturbance. Methods: This cross-sectional study used data from the UK Biobank: a cohort study of 502 655 participants, of whom 498 208 provided self-reported data on smoking and sleep characteristics. Multivariable multinomial and logistic regression models were used to examine the associations between smoking and sleep disturbance. Results: Long-sleep duration (>9 h) was more common among current smokers [odds ratio (OR): 1.47; 95% confidence interval (CI): 1.17-1.85; probability value (P) = 0.001] than never smokers, especially heavy (>20/day) smokers (OR: 2.85; 95% CI: 1.66-4.89; P < 0.001). Former heavy (>20/day) smokers were also more likely to report short (<6 h) sleep duration (OR: 1.41; 95% CI: 1.25-1.60; P < 0.001), long-sleep duration (OR: 1.99; 95% CI: 1.47-2.71; P < 0.001) and sleeplessness (OR: 1.47; 95% CI: 1.38-1.57; P < 0.001) than never smokers. Among never smokers, those who lived with more than one smoker had higher odds of long-sleep duration than those not cohabitating with a smoker (OR: 2.71; 95% CI: 1.26-5.82; P = 0.011). Conclusions: Active and passive exposure to high levels of tobacco smoke are associated with sleep disturbance. Existing global tobacco control interventions need to be enforced.

The effects of photic and nonphotic stimuli in the 5-HT7 receptor knockout mouse.

5-HT and agonists of the 5-HT receptor can modify the response of the mammalian pacemaker, which is located in the hypothalamic suprachiasmatic nuclei (SCN), to photic and nonphotic stimulation. Previous studies suggest that the 5-HT7 receptor is involved in the regulation of photic input, and the modulation of nonphotic circadian resetting of the circadian clock. The present study investigated the role of the 5-HT7 receptor by evaluating a wide variety of circadian parameters in mice lacking a functional allele for this receptor (5-HT7 knockout (KO)) compared with wild type (WT) animals that were bred on the same genetic background, including rate of entrainment, photic responsiveness and nonphotic response to a serotonergic agonist. No significant differences were detected in the average number of days 5-HT7 KO mice needed to reach entrainment to an advance of 6 h in the LD cycle compared with WT animals. Further, we investigated the acute responsiveness of both groups of mice to acute light stimulation at various times (circadian time (CT) 0, 6, 9, 12, 14, 16, 18, 20 and 22). A significant difference in the phase resetting response to light between the groups was revealed at CT22. Finally, as the 5-HT7 receptor has been associated with the modulation of nonphotic resetting in vitro, we examined the response of the 5-HT7 KO mice to systemic administration of a 5-HT(1A/7) agonist. The current study is the first to demonstrate the elimination of a nonphotic response to (+) 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) in mice lacking the 5-HT7 receptor compared with WT animals in vivo. Taken together, the present findings provide additional evidence that reform the established view on the role of the 5-HT7 in the photic regulation of retinohypothalamic (RHT) input, and support further the involvement of the 5-HT7 receptor in the modulation of nonphotic resetting in circadian clock.

Circadian rhythms of melatonin and behaviour in juvenile sheep in field conditions: Effects of photoperiod, environment and weaning.

Entrainment of circadian rhythms (CR) to the light dark cycle has been well described under controlled, experimental conditions. However, studies in rodents have reported that rhythms in the laboratory are not always reproduced under field conditions. The aim of this study was to characterise the CR of sheep maintained under conditions of standard UK farm animal husbandry and to investigate the effects of environmental challenges presented by season, weaning and changes in housing on CR. Male sheep (n = 9) were kept at pasture, or group housed in barns, under natural photoperiod for one year. CR in locomotor activity were monitored using accelerometry, and 24 h patterns in plasma cortisol and melatonin were measured every 4 h by ELISA. CR was measured before and after weaning, in summer and winter, and at pasture and by barn housing. Cosinor analysis revealed high amplitude, diurnal rhythms in locomotor activity that were disrupted by weaning and by barn housing. Rhythms in winter showed an interrupted night time activity pattern, but only when the sheep were kept at pasture. Cortisol and melatonin secretion followed typical circadian patterns in winter and summer. The CR of the sheep under the field conditions of this study were strikingly robust under basal conditions, but easily disrupted by environmental challenges. Interrupted patterns of activity during the long nights of wintertime, not previously reported for sheep kept in experimental conditions were recorded. Based on these findings, we propose that animals require exposure to more complex environments than the laboratory in order to exhibit their true circadian phenotype.

Genetic knockout and pharmacological blockade studies of the 5-HT7 receptor suggest therapeutic potential in depression.

The affinity of several antidepressant and antipsychotic drugs for the 5-HT7 receptor and its CNS distribution suggest potential in the treatment of psychiatric diseases. However, there is little direct evidence of receptor function in vivo to support this. We therefore evaluated 5-HT7 receptors as a potential drug target by generating and assessing a 5-HT7 receptor knockout mouse. No difference in assays sensitive to potential psychotic or anxiety states was observed between the 5-HT7 receptor knockout mice and wild type controls. However, in the Porsolt swim test, 5-HT7 receptor knockout mice showed a significant decrease in immobility compared to controls, a phenotype similar to antidepressant treated mice. Intriguingly, treatment of wild types with SB-258719, a selective 5-HT7 receptor antagonist, did not produce a significant decrease in immobility unless animals were tested in the dark (or active) cycle, rather than the light, adding to the body of evidence suggesting a circadian influence on receptor function. Extracellular recordings from hypothalamic slices showed that circadian rhythm phase shifts to 8-OH-DPAT are attenuated in the 5-HT7 receptor KO mice also indicating a role for the receptor in the regulation of circadian rhythms. These pharmacological and genetic knockout studies provide the first direct evidence that 5-HT7 receptor antagonists should be investigated for efficacy in the treatment of depression.

MDMA alters the response of the mammalian circadian clock in hamsters: effects on re-entrainment and triazolam-induced phase shifts.

Serotonin (5-hydroxytryptamine or 5-HT) is a neurotransmitter that is involved in a wide range of behavioural and physiological processes. Previous work has indicated that serotonin is important in the regulation of the circadian clock, which is located in the suprachiasmatic nuclei (SCN) of the hypothalamus. 3,4-methylenedioxymethamphetamine (MDMA or 'Ecstasy'), which is widely used as a recreational drug of abuse, is a serotonin neurotoxin in animals and non-human primates. Previous work has shown that MDMA exposure can alter circadian clock function both in vitro and in vivo. Evidence shows that 5-HT may have a modulatory role in the regulation of the circadian clock by non-photic stimuli, such as the benzodiazepine triazolam (TRZ). Triazolam is a short-acting benzodiazepine that results in phase advances of the wheel running activity in hamsters when administered during the mid-subjective day. In the present study, male Syrian hamsters treated with TRZ (5 mg/kg) at ZT6 significantly phase advanced their clock. Treatment with MDMA significantly diminished the TRZ induced phase shift in hamsters. Previous evidence shows the involvement of 5-HT in the re-synchronisation of the endogenous clock to a new shifted light-dark cycle. Untreated animals were successfully entrained to a new, 6 h advanced light-dark cycle within an average of 4.5 +/- 0.1 days. Following treatment with MDMA, these animals took an average of 8.3 +/- 0.1 days to re-entrain to a shifted environmental cycle. Immunohistochemical analysis revealed that animals treated with MDMA showed reduced serotonin staining, as evidenced by a decrease in innervation density in the SCN. No significant differences were found in cell counts within the raphe nuclei. These results demonstrate the importance of the serotonergic system in the modulation of photic and non-photic responses of the circadian pacemaker.

Phase response curves to neuropeptide Y in wildtype and tau mutant hamsters.

Neuropeptide Y (NPY)-containing fibers project from the intergeniculate leaflet to the suprachiasmatic nucleus. NPY has been shown to phase shift the circadian locomotor activity rhythm of wildtype hamsters, producing large phase advances in the subjective day and small delays in the subjective night. Previous studies have implicated this pathway in the mediation of activity-induced resetting of the circadian clock. Homozygous tau mutant and wildtype hamsters respond very differently to pulses of activity. Not only is the amplitude of the phase response curve exaggerated in the mutants with shifts of up to 7 h, but the stimuli are effective at different times during the cycle. Homozygous tau mutant hamsters and wildtype controls were implanted with guide cannulas aimed at the suprachiasmatic nucleus and injected with NPY at various times during the circadian cycle. The responses of homozygous tau mutant hamsters to NPY resembled their responses to nonphotic stimuli in both timing and direction of phase shift. This finding provides correlational evidence that NPY is involved in the effects of nonphotic behavioral events on the circadian system.

Attenuation of circadian light induced phase advances and delays by neuropeptide Y and a neuropeptide Y Y1/Y5 receptor agonist.

Circadian rhythms can be synchronised to photic and non-photic stimuli. The circadian clock, anatomically defined as the suprachiasmatic nucleus in mammals, can be phase shifted by light during the night. Non-photic stimuli reset the circadian rhythm during the day. Photic and non-photic stimuli have been shown to interact during the day and night. Precise mechanisms for these complex interactions are unknown. A possible pathway for non-photic resetting of the clock is thought to generate from the intergeniculate leaflet, which conveys information to the suprachiasmatic nucleus (SCN) through the geniculohypothalamic tract and utilises neuropeptide Y (NPY) as its primary neurotransmitter. Interactions between light and NPY were investigated during the early (2 h after activity onset) and late (6 h after activity onset) night in male Syrian hamsters. NPY microinjections into the region of the SCN significantly attenuated light-induced phase delay, during the early subjective night. Phase advances to light were completely inhibited by the administration of NPY during the late night. The precise mechanism by which NPY attenuates or blocks photic phase shifts is unclear, but the NPY Y5 receptor has been implicated in the mediation of this inhibitory effect. The NPY Y1/Y5 receptor agonist, [Leu(31),Pro(34)]NPY, was administered via cannula microinjections following light exposure during the early and late night. [Leu(31),Pro(34)]NPY significantly attenuated phase delays to light during the early night and blocked phase advances during the late night, in a manner similar to NPY. These results show the ability of NPY to attenuate phase shifts to light during the early night and block light-induced phase advances during the late night. Furthermore, this is the first in vivo study implicating the involvement of the NPY Y1/Y5 receptors in the complex interaction of photic and non-photic stimuli during the night. The alteration of photic phase shifts by NPY may influence photic entrainment within the circadian system.

Neuropeptide Y, GABA and circadian phase shifts to photic stimuli.

Circadian rhythms can be phase shifted by photic and non-photic stimuli. The circadian clock, anatomically defined as the suprachiasmatic nucleus (SCN), can be phase delayed by light during the early subjective night and phase advanced during the late subjective night. Non-photic stimuli reset the clock when presented during the subjective day. A possible pathway for the non-photic resetting of the clock is thought to originate from the intergeniculate leaflet, which conveys information to the SCN through the geniculohypothalamic tract and utilizes among others neuropeptide Y (NPY) and GABA as neurotransmitters. Photic and non-photic stimuli have been shown to interact during the early and late subjective night. Microinjections of NPY or muscimol, a GABA(A) receptor agonist, into the region of the SCN can attenuate light-induced phase shifts during the early and late subjective night. The precise mechanism for these interactions is unknown. In the current study we investigate the involvement of a GABAergic mechanism in the interaction between NPY and light during the early and late subjective night. Microinjections of NPY significantly attenuated light-induced phase delays and inhibited phase advances (P<0.05). The administration of bicuculline during light exposure, before NPY microinjection did not alter the ability of NPY to attenuate light-induced phase delays and block photic phase advances. These results indicate that NPY attenuates photic phase shifts via a mechanism independent of GABA(A) receptor activation. Furthermore it is evident that NPY influences circadian clock function via differing cellular pathways over the course of a circadian cycle.

Neuropeptide Y and behaviorally induced phase shifts.

Neuropeptide Y-containing fibers project from the intergeniculate leaflet of the lateral geniculate nucleus to the suprachiasmatic nucleus. Previous studies have indicated that this pathway may be involved in non-photic resetting of the circadian clock. Therefore, we investigated the possibility that neuropeptide Y mediates phase shifts induced by a particular non-photic stimulus, a pulse of running in a novel wheel. Confining hamsters to a small nest box failed to block phase shifts induced by neuropeptide Y given at zeitgeber time 4; this indicates that increased locomotor activity is not necessary for the observed shifts. Antiserum raised against neuropeptide Y or normal serum was administered at circadian time 5 through a cannula aimed at the suprachiasmatic nucleus. The hamsters were then removed from their cages and placed in a novel wheel for 3 h. Hamsters that received normal serum and ran > 5000 revolutions in the novel wheel advanced their rhythms (mean shift 2.55 h +/- 0.22 S.E.M.) by amounts similar to those of unoperated hamsters. Administration of neuropeptide Y antiserum attenuated the shift normally associated with running in a novel wheel (mean shift 0.21 h +/- 0.14 S.E.M.). These studies indicate that the neuropeptide Y input from the lateral geniculate nucleus to the biological clock is involved in the phase shifts seen in response to novelty-induced wheel running. It also provides another example of the ability of antisera to alter behavior. This may be a useful approach in manipulations of neurochemical activity when antagonists are not yet available or poorly defined.

Neuropeptide Y and glutamate block each other's phase shifts in the suprachiasmatic nucleus in vitro.

The suprachiasmatic nuclei contain a circadian clock whose activity can be recorded in vitro for several days. Photic information is conveyed to the nuclei primarily via a direct projection from the retina, the retinohypothalamic tract, utilizing an excitatory amino acid neurotransmitter. Photic phase shifts may be mimicked by application of glutamate in vitro. A second, indirect pathway to the suprachiasmatic nuclei via the geniculohypothalamic tract utilizes neuropeptide Y as a transmitter. Phase shifts to neuropeptide Y in vitro are similar to those seen to non-photic stimuli in vivo. We have used the hypothalamic slice preparation to examine the interactions of photic and non-photic stimuli in the suprachiasmatic nuclei. Coronal hypothalamic slices containing the suprachiasmatic nuclei were prepared from Syrian hamsters and 3 min recordings of the firing rate of individual cells were performed throughout a 12 h period. Control slices receiving either no application or application of artificial cerebrospinal fluid to the suprachiasmatic nucleus showed a consistent daily peak in their rhythms. Glutamate produces phase shifts of the circadian clock in the hamster hypothalamic slice preparation during the subjective night but not during the subjective day. These phase shifts were similar in timing and direction to the photic phase response curve in vivo confirming previous work with the rat slice preparation. Neuropeptide Y produces phase shifts of the circadian clock during the subjective day but not during the subjective night. The phase shifts are similar in timing and direction to the non-photic phase response curve in vivo, confirming previous in vitro work. We then examined the interaction of these neurochemicals with each other at various times during the circadian cycle. We found that both advances and delays to glutamate in the slice are blocked by application of neuropeptide Y. We also found that phase shifts to neuropeptide Y in the slice are blocked by application of glutamate. These results indicate that photic and non-photic associated neurochemicals can block each others phase shifting effects within the suprachiasmatic nucleus in vitro. These experiments demonstrate the ability of photic and non-photic associated neurochemicals to interact at the level of the suprachiasmatic nucleus. It is clear that neuropeptide Y antagonizes the effect of glutamate during the subjective night, and that glutamate antagonizes the effect of neuropeptide Y during the subjective day. Great care must be taken when devising treatments where photic and non-photic signals may interact.

Blocking the phase-shifting effect of neuropeptide Y with light.

Previous studies have indicated that the neuropeptide Y input from the intergeniculate leaflet of the lateral geniculate nucleus to the suprachiasmatic nucleus is the final part of a non-photic phase shifting pathway to pacemakers in hamsters, or that neuropeptide Y is necessary for other pathways to be effective. Experiments in which two stimuli are presented during the same circadian cycle have shown that phase shifts in response to at least two non-photic stimuli are attenuated by a subsequent light pulse during the subjective day. This study was conducted to investigate the neural site of the blocking effect of light on non-photic stimuli. Experiment 1 showed that phase shifts in response to induced wheel-running during the subjective day are greatly attenuated by a subsequent light pulse. Experiment 2 showed that phase shifts in response to injections of neuropeptide Y in the middle of the subjective day were also greatly reduced by a subsequent light pulse. These results provide some insight about the site of the blocking action of light on non-photic phase shifts. Because there is evidence indicating that neuropeptide Y may mediate phase shifts in response to induced activity, and because light was able to block phase shifts produced by neuropeptide Y, we conclude that, in blocking activity-induced shifts, light must act downstream from the release of neuropeptide Y into the suprachiasmatic nucleus.

Neuropeptide Y phase shifts the circadian clock in vitro via a Y2 receptor.

The suprachiasmatic nuclei (SCN) contain a circadian clock whose activity can be recorded in vitro for several days. This clock can be reset by the application of neuropeptide Y. In this study, we focused on determination of the receptor responsible for neuropeptide Y phase shifts of the hamster circadian clock in vitro. Coronal hypothalamic slices containing the SCN were prepared from Syrian hamsters housed under a 14 h:10 h light:dark cycle. Tissue was bathed in artificial cerebrospinal fluid (ACSF), and the firing rates of individual cells were sampled throughout a 12 h period. Control slices received either no application or application of 200 nl ACSF to the SCN at zeitgeber time 6 (ZT6; ZT12 was defined as the time of lights off). Application of 200 ng/200 nl of neuropeptide Y at ZT6 resulted in a phase advance of 3.4 h. Application of the Y2 receptor agonist, neuropeptide Y (3-36), induced a similar phase advance in the rhythm, while the Y1 receptor agonist, [Leu31, Pro34]-neuropeptide Y had no effect. Pancreatic polypeptide (rat or avian) also had no measurable phase-shifting effect. Neuropeptide Y applied at ZT20 or 22 had no detectable phase-shifting effect. These results suggest that the phase-shifting effects of neuropeptide Y are mediated through a Y2 receptor, similar to results found in vivo.

Enhanced photic phase shifting after treatment with antiserum to neuropeptide Y.

Previous work has shown that antiserum to neuropeptide Y blocks phase shifts to certain non-photic stimuli at circadian time 4. To examine the role of NPY in photic phase shifts, antiserum to neuropeptide Y was administered just prior to a light pulse at circadian time 18. Hamsters were implanted with guide cannulae aimed at the suprachiasmatic nucleus, and housed in constant darkness. Animals were injected at circadian time 18 under a dim red safe light with 200 nl of either antiserum to neuropeptide Y or normal serum. They were then either exposed to light (approximately 100 lux) for 15 min or returned to constant darkness. Hamsters shifted more to the light pulse when pretreated with antiserum to neuropeptide Y (mean normal serum + light = 1.18 h: mean antiserum to neuropeptide Y + light = 1.92 h; P < 0.01 on a two-tailed paired t-test). Therefore, antiserum to neuropeptide Y enhanced photic advances at circadian time 18.

Circadian phase-shifts induced by chlordiazepoxide without increased locomotor activity.

Phase advances of circadian locomotor rhythms in response to the benzodiazepine triazolam (TRZ) administered at circadian time 6 appear to be mediated by locomotor activity. The possibility that increased activity also mediates shifts induced by the benzodiazepine chlordiazepoxide (CDZ) was investigated. Hamsters injected with CDZ 7 h before dark onset exhibited phase advances despite being confined to their nest boxes for 3 h after receiving the drug. These shifts did not differ significantly from advances seen in hamsters which were allowed access to the rest of the cage after injections. In other experiments, the behavioral effects of both CDZ and TRZ were examined during the 5 h after injection. There was significantly more motor activity observed after injection of TRZ than CDZ. These results suggest that phase-shifting effects of some benzodiazepines can occur without inducing activity.

MDMA and fenfluramine alter the response of the circadian clock to a serotonin agonist in vitro.

The substituted amphetamine drugs, 3,4-methylenedioxymethamphetamine (MDMA or 'Ecstasy') and fenfluramine, are known to damage 5-HT neurons in the brain of animals. However, little is known about the drugs' effects on circadian rhythmicity which is known to be influenced by serotonergic input to the suprachiasmatic nuclei. In the present study, we tested the ability of MDMA and fenfluramine treatment to alter the ability of the circadian clock to reset in response to an agonist of the 5-HT1A and 5-HT7 receptor subtypes soon after treatment with the drugs, and then again at 20 weeks. Coronal hypothalamic slices containing the suprachiasmatic nuclei (SCN) were prepared from rats and 3-min recordings of the firing rate of individual cells were performed throughout a 12-h period. The ability of the 5-HT agonist, 8-hydroxy-2-(dipropylamino)tetralin (8-OH-DPAT), to cause a phase advance in the firing pattern of SCN neurons was assessed in slices from control animals and those pretreated with MDMA or fenfluramine (10, 15 and 20 mg/kg administered on successive days) 6-10 days or 20 weeks previously. Phase advances to 8-OH-DPAT in the slice were attenuated by pretreatment with MDMA or fenfluramine at both drug-test intervals. Our study demonstrates that repeated exposure to MDMA or fenfluramine may interfere with the ability of serotonin to phase shift the circadian clock in the rat. It is possible that such an effect may be responsible for some of the clinical changes, such as sleep disorders and mood changes, sometimes reported by human users of the substituted amphetamines.

Geniculo-hypothalamic tract lesions block chlordiazepoxide-induced phase advances in Syrian hamsters.

Administration of the benzodiazepine triazolam at the appropriate time in the circadian cycle has been shown to induce phase shifts in hamster circadian rhythms. These phase shifts can be blocked by geniculo-hypothalamic tract (GHT) ablation or by restraint of activity. The present study examined the effects of the benzodiazepine chlordiazepoxide on running-wheel activity rhythms of hamsters. The phase-advancing effect of intraperitoneal injections of chlordiazepoxide administered at circadian time 6 (CT 6) was dose-dependent. Average shifts ranged from 6 min at a dose of 0.05 mg/kg to 135 min at a dose of 200 mg/kg. Four of twenty hamsters did not show a phase shift to any dose tested. Phase advance shifts to chlordiazepoxide (CT 6; 100 mg/kg) were blocked by GHT lesions. Chlordiazepoxide injections at doses which induced phase shifts were often followed by sedation. These results indicate that chlordiazepoxide is similar to triazolam, in that its ability to induce phase shifts at circadian time 6 is blocked by GHT lesions.

The effects of GABA and benzodiazepines on neurones in the suprachiasmatic nucleus (SCN) of Syrian hamsters.

Administration of benzodiazepines at appropriate times in the circadian cycle induce phase-shifts in circadian locomotor activity. The possibility that benzodiazepine-induced shifts are mediated at the level of the suprachiasmatic nuclei (SCN), identified as the circadian pacemaker in mammals, was examined electrophysiologically. Extracellular recordings were made from Syrian hamster (Mesocricetus auratus) hypothalamic SCN neurones in vitro to assess (1) the effects of gamma-aminobutyric acid (GABA) on SCN neuronal activity and (2) the effects of benzodiazepines (chlordiazepoxide and flurazepam) on GABA-evoked responses. Of 93 SCN cells tested, 86 were suppressed by iontophoresed GABA (20 mM) in a current(dose)-dependent manner, while 6 were unaffected; suppression was found during both the projected light and dark phases of the circadian cycle. Application of bicuculline methiodide alone elevated mean discharge activity, while GABA-evoked suppressions were blocked by bicuculline (n = 9/11 cells). Iontophoresis of chlordiazepoxide or flurazepam (20 mM; 1-10 nA) alone produced a current(dose)-dependent prolonged suppression of cell firing which was antagonised by bicuculline. These results indicate that benzodiazepine/GABA-evoked responses are at least partially mediated by GABAA receptors within the SCN and suggest that SCN may be a possible locus for the action of benzodiazepines in their induction of phase-shifts in circadian function.

A neurophysiological study of a lithium-sensitive phosphoinositide system in the hamster suprachiasmatic (SCN) biological clock in vitro.

Lithium lengthens the period of free-running circadian rhythms in many species. In mammals the hypothalamic suprachiasmatic nucleus (SCN) has been identified as a biological clock which generates circadian rhythms. The effect of lithium-induced depletion of the intracellular pool of inositol, leading to decreased intracellular second messengers IP3 and DAG, was examined neurophysiologically. Extracellular recordings were obtained from spontaneously discharging SCN neurones maintained in vitro. Superfusion of slices with lithium-containing (0.1-30 mM) aCSF, but not rubidium-containing aCSF, suppressed neuronal firing in a dose-dependent manner. Lithium-induced suppressed firing was reversed by myo-inositol, but not by epi-inositol. These studies provide evidence for basal phosphoinositide turnover in neurones and implicate a lithium-sensitive phosphoinositide system in the maintenance of the spontaneous discharge activity of SCN neurones.

WAY-100635, a specific 5-HT1A antagonist, can increase the responsiveness of the mammalian circadian pacemaker to photic stimuli.

Mammalian circadian rhythms are synchronized daily to light-dark cycles in the environment. The suprachiasmatic nucleus (SCN) is the proposed site of the major circadian pacemaker. Daily entrainment is believed to be influenced by inputs to the SCN, one of these being the dense serotonergic (5-HT) projection from the raphe nuclei. WAY-100635 is a potent and selective 5-HT1A receptor antagonist. In this study, the effects of WAY-100635 on phase-shifts of the hamster circadian pacemaker to light were investigated. Phase-delays after a light pulse administered during the early subjective night (15 min at CT14) were observed to be significantly greater following pre-treatment with WAY-100635 compared to light pulse alone (P < 0.05). However, pre-treatment with WAY-100635 had no effect on the magnitude of phase-shifts to light at CT18, late in the subjective night. Serotonin may influence the responsiveness of the circadian pacemaker to photic stimuli. Specifically, WAY-100635 administered at CT14 can augment phase-shifts to light.

Nonphotic phase shifting in the old and the cold.

When confined to novel running wheels or when given injections of triazolam in their home cages, old hamsters do not become as active as young hamsters. Therefore, lack of nonphotic phase shifting following such manipulations may stem from insufficient activity or arousal. Phase advances can be obtained in some 10-month-old animals when wheel running during the pulse is increased by the presence of females in estrous condition and in most 18-month-old hamsters by combining confinement to a novel wheel with triazolam injections. These data suggest that there is relatively little if anything wrong in aging hamsters with the nonphotic phase-shifting mechanism itself. The reason why in certain situations old hamsters do not shift appears to be because the nonphotic inputs to these shifting mechanisms are not strong enough. However, when running in novel wheels is increased by carrying out the tests at cold temperatures, most old animals did not show subsequent phase shifts. Evidently it is not running per se that is critical for phase shifts, but probably the motivational context for such running.