Circadian clock speed increases during aging in the male Syrian hamster: A large-scale study.

There are conflicting reports in the literature as to whether or not the circadian period in Syrian hamsters shortens with age, and those studies were conducted with small sample sizes. This report mines data from a large number of experiments resulting in more than 1000 measurements of circadian period in hamsters during aging up to 6 months, and this was correlated with both age and weight. Circadian period was calculated while hamsters were in running wheels in constant darkness. There is a weak correlation between hamster weight and circadian period, and there is significant shortening of circadian period during aging.

Non-peptide oxytocin receptor ligands and hamster circadian wheel running rhythms.

The synchronization of circadian rhythms in sleep, endocrine and metabolic functions with the environmental light cycle is essential for health, and dysfunction of this synchrony is thought to play a part in the development of many neurological disorders. There is a demonstrable need to develop new therapeutics for the treatment of neurological disorders such as depression and schizophrenia, and oxytocin is currently being investigated for this purpose. There are no published reports describing activity of oxytocin receptor ligands on mammalian circadian rhythms and that, then, is the purpose of this study. Non-peptide oxytocin receptor ligands that cross the blood brain barrier were systemically injected in hamsters to determine their ability to modulate light-induced phase advances and delays of circadian wheel running rhythms. The oxytocin receptor agonist WAY267464 (10 mg/kg) inhibited light induced phase advances of wheel running rhythms by 55%, but had no effect on light-induced phase delays. In contrast, the oxytocin receptor antagonist WAY162720 (10 mg/kg) inhibited light-induced phase delays by nearly 75%, but had no effect on light-induced phase advances. Additionally, WAY162720 was able to antagonize the inhibitory effects of WAY267464 on light-induced phase advances. These results are consistent for a role of oxytocin in the phase-delaying effects of light on circadian activity rhythms early in the night. Therefore, oxytocin may prove to be useful in developing therapeutics for the treatment of mood disorders with a concomitant dysfunction in circadian rhythms. Investigators should also be cognizant that oxytocin ligands may negatively affect circadian rhythms during clinical trials for other conditions.

Effects of systemically applied nAChRα7 agonists and antagonists on light-induced phase shifts of hamster circadian activity rhythms.

Many physiological systems in mammals are linked to the body's master circadian rhythm in the sleep/wake cycle and dysfunctions in this rhythm has been associated with neurological diseases such as major depression, Alzheimer's Disease and schizophrenia. There is some evidence that nicotinic cholinergic input to the master circadian pacemaker, the suprachiasmatic nucleus, may modulate circadian activity rhythms, but data employing in vivo preparations is sparse. Therefore we examined the ability of intraperitoneally applied nicotinic agonists and antagonists relatively selective for the α7 nicotinic receptor to modulate light-induced phase shifts of hamster circadian wheel running rhythms. Hamsters were maintained in constant darkness and exposed to light pulses early and late in their active period, mimicking dusk and dawn respectively, which elicited phase delays and advances of their circadian wheel running rhythms. The α7 receptor antagonists bPiDDB (0.03-3mg/kg) and methyllacaconitine (0.1-1mg/kg) inhibited both light- induced phase advances and delays of circadian wheel running rhythms by as much as 75% versus vehicle injections. In contrast, systemic injections of the α7 agonists PHA 543613 and ABT107, both at 0.156-2.5mg/kg, had no effect on light induced phase advances or delays. Further, α7 nicotinic receptors were identified in the hamster suprachiasmatic nucleus using an antibody that recognizes α7 nicotinic receptors. These results clearly identify the ability of α7 nicotinic receptor antagonists to inhibit light-entrainment of the hamster circadian pacemaker. Therefore, nicotinic compounds may be useful for the treatment of circadian dysfunction associated with neurological diseases.

OPSITE Flexifix Gentle: preventing breakdown in vulnerable skin.

For decades, the intact skin of patients has been at risk of damage from exposure to pressure or friction from devices used to support their treatment while in hospital. Device-related skin injury is a common cause of skin breakdown in the acute hospital environment. The use of clinical devices exposes patients to repetitive friction and light pressure that, if not recognised early, can lead to skin breakdown. Compounding this challenge of protecting skin is the use of adhesive tape that can cause skin stripping. A new dressing product called OPSITE Flexifix Gentle (OFG) is now available on the market. This product may support friction relief in these critical areas and prevent skin stripping and pressure ulcer formation while maintaining moisture balance and a healthy skin environment. This product focus outlines the clinical benefits from this silicone-based film roll. The performance of the product on a 34-bed cardiothoracic intensive care unit will be reviewed through three case studies.

LY2033298, a positive allosteric modulator at muscarinic M4 receptors, enhances inhibition by oxotremorine of light-induced phase shifts in hamster circadian activity rhythms.

RATIONALE: Entrainment of circadian rhythms to the light-dark cycle is essential for restorative sleep, and abnormal sleep timing is implicated in central nervous system (CNS) disorders like depression, schizophrenia, and Alzheimer's disease. Many transmitters, including acetylcholine, that exerts its actions via muscarinic receptors modulate the suprachiasmatic nucleus, the master pacemaker. OBJECTIVES: Since positive allosteric modulators of muscarinic M(4) receptors are candidates for treatment of mood and cognitive deficits of CNS disorders, it is important to evaluate their circadian actions. MATERIALS AND METHODS: The effects of intraperitoneally applied muscarinic agents on circadian wheel-running rhythms were measured employing hamsters, a model organism for studying activity rhythms. RESULTS: Systemic administration of the muscarinic receptor agonist oxotremorine (0.01-0.04 mg/kg) inhibited light-induced phase delays and advances of hamster circadian wheel-running rhythms. The M4 positive allosteric modulator, LY2033298 (10-40 mg/kg), had no effect on light-induced phase shifts when administered alone, yet significantly enhanced (at 20 mg/kg) the inhibitory influence of oxotremorine on light-induced phase delays. In addition, the muscarinic receptor antagonist, scopolamine, which was without effect on light-induced phase shifts when administered alone (0.001-0.1 mg/kg), antagonized (at 0.1 mg/kg) the inhibitory effect of oxotremorine and LY2033298 on light-induced phase delays. CONCLUSIONS: These results are the first to demonstrate that systemically applied muscarinic receptor agonists modulate circadian activity rhythms, and they also reveal a specific role for M4 receptors. It will be of importance to evaluate circadian actions of psychotropic drugs acting via M4 receptors, since they may display beneficial properties under pathological conditions.

Positive allosteric modulators at GABAB receptors exert intrinsic actions and enhance the influence of baclofen on light-induced phase shifts of hamster circadian activity rhythms.

Light-induced phase shifts of hamster circadian activity rhythms are modulated by GABA(B) receptors. Recently, positive allosteric modulators (PAM)s at GABA(B) receptors were described, but it is not known whether they affect light-induced entrainment of circadian rhythms. Therefore, we studied the effects of two GABA(B) PAMs, GS39783 and RacBHFF, upon light-induced phase advances and delays of hamster circadian wheel-running activity rhythms. Wheel running activity was recorded for Syrian hamsters maintained in constant darkness. Drugs administered intraperitoneally were evaluated for their ability to modulate a light-induced shift of the circadian activity rhythm. Baclofen (3.75-15 mg/kg) dose-dependently inhibited both light-induced phase advances and delays of hamster wheel running rhythms, and its actions were blocked by the selective GABA(B) antagonist, SCH50911 (5mg/kg). Neither GS39783 (3-30 mg/kg) nor RacBHFF (0.63-10mg/kg) affected phase advances when injected alone, but both GS39783 (3mg/kg) and RacBHFF (10mg/kg) augmented the inhibitory effect of baclofen (5mg/kg). At doses above 3mg/kg, GS39783 and RacBHFF significantly inhibited phase delays alone, consistent with the notion of "agonist-allosteric" properties. GS39783 (0.5mg/kg), but not RacBHFF (10mg/kg), augmented the inhibitory action of baclofen on phase delays. These data are consistent with the possibility that GS39783 and RacBHFF act as PAMs at GABA(B) receptors inhibiting light-induced phase advances, yet that they also posses "allosteric agonist" actions at the (presumably separate) population of GABA(B) receptors modulating light-induced phase delays. GABA(B) receptors clearly warrant further investigation as agents for modulation of circadian dysfunction associated with CNS disorders such as depression.

Positive and negative modulation of circadian activity rhythms by mGluR5 and mGluR2/3 metabotropic glutamate receptors.

Glutamate released from retinal ganglion cells conveys information about the daily light:dark cycle to master circadian pacemaker neurons within the suprachiasmatic nucleus that then synchronize internal circadian rhythms with the external day-length. Glutamate activation of ionotropic glutamate receptors in the suprachiasmatic nucleus is well established, but the function of the metabotropic glutamate receptors that are also located in this nucleus is not known. Therefore, in this study we evaluated agonists and antagonists acting at orthosteric or allosteric sites for mGluR5 and mGluR2/3 metabotropic glutamate receptors for their ability to modulate light-induced phase advances and delays of hamster circadian activity rhythms. mGluR5 allosteric antagonists fenobam, MPEP and MTEP, each 10 mg/kg, potentiated light-induced phase advances of hamster circadian activity rhythms, while the mGluR5 agonists CHPG, (S)-3,5-DHPG or positive allosteric modulator CDPPB had no effect. Neither mGluR5 agonists nor antagonists had any effect on light-induced phase delays of activity rhythms. The competitive mGluR2/3 antagonist LY341495, 10 mg/kg, also potentiated light-induced phase advances, but inhibited light-induced phase delays. The mGluR2/3 agonists LY354740 and LY404039 were without effect on phase advances while a third agonist LY379268, 10 mg/kg, inhibited both light-induced advances and delays. Finally, mGluR2/3 agonists LY379268 and LY404039 also inhibited light-induced phase delays of activity rhythms. These results suggest that during light-induced phase advances, mGluR2/3 and mGluR5 receptors act to negatively modulate the effects of light on the circadian pacemaker or its output(s). mGluR5 receptors do not appear to be involved during light-induced phase delays. In contrast, the role for mGluR2/3 receptors during phase delays is more complicated as both agonists and antagonists inhibit light-induced phase delays. Dysfunctions in human circadian rhythms have been implicated in some forms of depression, and metabotropic glutamate receptor ligands, which are also being evaluated for antidepressant activity, are shown here to be capable of modifying light-induced phase shifts of circadian activity rhythms.

The benzodiazepine diazepam demonstrates the usefulness of Syrian hamsters as a model for anxiety testing: evaluation of other classes of anxiolytics in comparison to diazepam.

Clinical evidence in humans suggests that there is some linkage between dysfunction in the timing of circadian rhythms and certain types of depression. In animal models, Syrian hamsters have been used extensively to study the pharmacology of circadian rhythms, while rats and mice are used to screen putative anxiolytics/antidepressant compounds. It would be beneficial to be able to test anxiolytic/antidepressant compounds in hamsters in conjunction with circadian rhythm studies. Therefore, in this study, Syrian hamsters were used in three experimental paradigms to evaluate anxiety: the elevated plus maze, the t-tube, and the open field Thatcher-Britton conflict test. Diazepam, tested with 2mg/kg and 5mg/kg intraperitoneal injections, was found to induce anxiolytic activity in each of the three tests. Hamsters were more likely to spend time in the open arms in the plus maze, displayed more exploratory behavior in the t-tube, and were quicker to enter a brightly lit exposed field in the Thatcher-Britton conflict test following injections of diazepam. Diazepam (2mg/kg) was also tested at three times during the 24-h day in the elevated plus maze: at the beginning and end of the lights-on period (Zeitgeber times 23 and 11, respectively) and once in the dark just before the room lights came on (Zeitgeber time 20). Diazepam induced anxiolytic activity only at Zeitgeber 23. Therefore, the following known and putative anxiolytic compounds were also evaluated in each of the three tests at Zeitgeber 23: citalopram, the neurokinin(1) receptor antagonists GR205171 and vestipitant, the corticotropin releasing factor(1) receptor antagonist CP154526, the cannabinoid receptor(1) agonist CP55940, the serotonin(6) receptor antagonist SB399885, and the metabotropic glutamate receptor(5) antagonists fenobam and MTEP. Vestipitant displayed some anxiolytic activity in the elevated plus maze, but this effect was not confirmed with GR205171. None of the other compounds displayed any anxiolytic activity in the tests. Nevertheless, the present results with diazepam - together with a few reports from other laboratories, indicate that the elevated plus maze may be a suitable procedure for evaluating the actions of anxiolytic compounds in Syrian hamsters. In view of current interest in novel classes of psychotropic agent interacting with diverse GABA(A) receptor recognition sites, further characterization appears justified.

Differential influence of selective 5-HT5A vs 5-HT1A, 5-HT1B, or 5-HT2C receptor blockade upon light-induced phase shifts in circadian activity rhythms: interaction studies with citalopram.

Though serotonergic mechanisms modulate circadian rhythms, roles of individual serotonin (5-HT) receptors remain uncertain since data are lacking for antagonists. Herein, both the 5-HT(5A) receptor antagonist, A843277 (10 mg/kg), and the 5-HT(1B) antagonist, SB224289 (1 mg/kg), inhibited light-induced phase advances in hamster circadian wheel-running rhythms. Conversely, though 5-HT(1A) and 5-HT(7) receptors are likewise implicated in circadian scheduling, their blockade by WAY100635 (0.5 mg/kg) and SB269970 (1 mg/kg), respectively, was ineffective. Since actions of 5-HT reuptake inhibitors are modified by antagonists, we evaluated their influence on suppression of phase advances by citalopram (10 mg/kg). Its action was potentiated by WAY100635 and the 5-HT(2C) antagonist, SB242084 (1 mg/kg), but not by A842377, SB224289, SB269970, and antagonists at 5-HT(2A) (MDL100907) and 5-HT(6) (SB399885) receptors. In conclusion, this is the first in vivo evidence for an influence of 5-HT(5A) receptors upon circadian rhythms, but no single class of 5-HT receptor mediates their control by citalopram.

Neurokinin1 antagonists potentiate antidepressant properties of serotonin reuptake inhibitors, yet blunt their anxiogenic actions: a neurochemical, electrophysiological, and behavioral characterization.

Though neurokinin(1) (NK(1)) receptor antagonists are active in experimental models of depression, clinical efficacy has proven disappointing. This encourages interest in association of NK(1) receptor blockade with inhibition of serotonin (5-HT) reuptake. The selective NK(1) antagonist, GR205171, dose-dependently enhanced citalopram-induced elevations of extracellular levels of 5-HT in frontal cortex, an action expressed stereospecifically vs its less active distomer, GR226206. Further, increases in 5-HT levels in dorsal hippocampus, basolateral amygdala, nucleus accumbens, and striatum were likewise potentiated, and GR205171 similarly facilitated the influence of fluoxetine upon levels of 5-HT, as well as dopamine and noradrenaline. In parallel electrophysiological studies, the inhibitory influence of citalopram and fluoxetine upon raphe-localized serotonergic neurones was stereospecifically blunted by GR205171. Antidepressant actions of citalopram in a forced-swim test in mice were stereospecifically potentiated by GR205171, and it also enhanced attenuation by citalopram of stress-related ultrasonic vocalizations in rats. Further, GR205171 and citalopram additively abrogated the advance in circadian rhythms provoked by exposure to light in hamsters. By contrast, GR205171 stereospecifically blocked anxiogenic actions of citalopram in social interaction procedures in rats and gerbils, and stereospecifically abolished facilitation of fear-induced foot tapping by fluoxetine in gerbils. By analogy to GR205171, a further NK(1) antagonist, RP67580, enhanced the influence of citalopram upon frontocortical levels of 5-HT and potentiated its actions in the forced swim test. In conclusion, NK(1)receptor blockade differentially modulates functional actions of SSRIs: antidepressant properties are reinforced, whereas anxiogenic effects are attenuated. Combined NK(1) receptor antagonism/5-HT reuptake inhibition may offer advantages in the management of depressed and anxious states.

Serotonin1A-mediated amplification of light-induced phase advances of circadian rhythms in the Syrian hamster: post-light effects.

Certain serotonergic compounds that have an agonist/antagonist activity at the 5-HT(1A) receptor subtype are known to greatly potentiate the ability of light to advance the phase of circadian activity rhythms in hamsters. Typically, investigational compounds are injected 30 min to 1 h prior to a phase-advancing light pulse, and it is not known if these compounds are efficacious when injected after a short light pulse. In this study we injected the 5-HT(1A) mixed agonist/antagonist BMY 7378 from 1 to 7 h after a short phase advancing light pulse at CT 19 in hamsters to assess the temporal window of efficacy of this compound. BMY 7378 effectively doubles the magnitude of light-induced phase advances in hamster circadian wheel running rhythms for 6 h after a light pulse administered at CT 19, and this effect abruptly ends at 7 h post-light. This demonstrates that the molecular events initiated by a light pulse at CT 19 responsible for initiating phase advances in circadian activity rhythms can be modulated for at least the next 6 h in the hamster. However, this effect found with BMY 7378 does not extend to all serotonergic compounds as the inhibitory activity of 8-OH-DPAT on light-induced phase advances is not apparent when injected just 3 h after a light pulse. Since BMY 7378 can elicit such major shifts in the timing of the circadian clock it should be useful as a tool to explore potential changes in gene expression that lead to phase shifts in circadian rhythms, now known to be susceptible to modification for 6 h after a phase-advancing light pulse.

Cannabinoids and hamster circadian activity rhythms.

Circadian activity rhythms in hamsters are entrained to the daily light:dark cycle by photic information arriving from the retina to the suprachiasmatic nucleus, the site of the master circadian pacemaker in mammals. The effects of light on adjusting the timing of the circadian pacemaker is modified, both positively and negatively, by a variety of transmitter systems, but the effects of endocannabinoids have not been reported. Therefore, in this study we evaluated cannabinoids specific for the cannabinoid type 1 receptor (CB(1)) for their ability to modulate light-induced phase advances in hamster circadian activity rhythms. All compounds were administered intraperitoneally. The CB(1) agonist CP55940 potently inhibited light-induced phase shifts with near 90% inhibition achieved with a dose of 0.125 mg/kg. The inhibitory effect of CP55940 was partially reversed by the CB(1) antagonist LY320135 and completely reversed with 1 mg/kg of the CB(1) antagonist AM 251. Neither LY320135 nor AM 251 had any effect on light-induced phase shifts when administered alone. Further evidence for CB(1) involvement in hamster circadian rhythms was provided by immunohistochemical detection of CB(1) receptors in four separate nuclei comprising the principal components of the hamster circadian system: the suprachiasmatic nucleus, intergeniculate leaflet of the thalamus, and dorsal and median raphe nuclei. Altogether these data indicate that the endocannabinoid system has the capability to modulate circadian rhythms in the hamster and cannabis use should be evaluated for adverse effects on circadian rhythms in humans.

Evaluation of serotonin, noradrenaline and dopamine reuptake inhibitors on light-induced phase advances in hamster circadian activity rhythms.

RATIONALE: Selective serotonin reuptake inhibitors (SSRIs) are widely prescribed for the treatment of anxiodepressive states that are often associated with perturbed circadian rhythms including, in certain patients, phase advances. Surprisingly, the influence of SSRIs upon circadian activity rhythms has been little studied in experimental models. OBJECTIVES: Accordingly, this study examined the ability of SSRIs to modulate the phase-setting properties of light on circadian activity rhythms in hamsters. Their actions were compared to those of the mixed serotonin/noradrenaline reuptake inhibitor (SNRI), venlafaxine, the selective noradrenaline reuptake inhibitor, reboxetine, and the dopamine reuptake inhibitor, bupropion. MATERIALS AND METHODS: Wheel-running activity rhythms were recorded in male Syrian hamsters. Drugs were administered systemically before a light stimulus that was used to advance the timing of the hamster running rhythms. RESULTS: Four chemically diverse SSRIs, citalopram (1-10 mg/kg, intraperitoneally), fluvoxamine (1-10), paroxetine (1-10), and fluoxetine (10 and 20), all robustly and significantly inhibited the ability of light to phase advance hamster circadian wheel-running activity rhythms. Their actions were mimicked by venlafaxine (1-10) that likewise elicited a marked reduction in phase advances. Conversely, reboxetine (1-20) and bupropion (1-20) did not exert significant effects. CONCLUSIONS: These data suggest that suppression of serotonin (but not noradrenaline or dopamine) reuptake by SSRIs and SNRIs modifies circadian locomotor activity rhythms in hamsters. Further, they support the notion that an inhibitory influence upon the early-morning light-induced advance in circadian activity contributes to the therapeutic effects of serotonin uptake inhibitors in certain depressed patients.

A review of the four layer vs the short stretch bandage system.

This article reviews current evidence in the treatment of venous leg ulcers. In particular, discussion focuses on whether the four layer bandage system (4LB) or the short stretch (SSB) system should be considered as the first-line treatment of venous leg ulcers. Using research terminology, relevant studies are analysed for academic metric. In addition, a concluding discussion is held to consider the practical benefits of selecting a bandage strategy.

Wound cleansing: sterile water or saline?

Robert Gannon discusses whether sterile 0.9% saline or sterile water should be used as the main cleansing solution in hospital settings. He also explores the importance of warming cleansing solutions before use. The physiological and practical benefits of each solution will be analysed.

The corticotropin-releasing factor (CRF)(1) receptor antagonists CP154,526 and DMP695 inhibit light-induced phase advances of hamster circadian activity rhythms.

The circadian activity of corticotropin releasing factor (CRF) and the hypothalamic-pituitary-adrenal axis is controlled by the master circadian pacemaker located in the hypothalamic suprachiasmatic nucleus. However, the reciprocal influence of CRF and the hypothalamic-pituitary-adrenal axis upon the circadian pacemaker is less well established. Therefore, in the present study, we tested two nonpeptidergic antagonists at CRF(1) receptors for their ability to modulate photic resetting of pacemaker time (phase). CP154,526 dose dependently and significantly inhibited light-induced phase advances in hamster circadian activity rhythms late in the subjective night by approximately 60% at a maximally effective dose of 20 mg/kg delivered intraperitoneally. Likewise, a further CRF(1) receptor antagonist, DMP695, inhibited phase advances by approximately 40% at a dose of 10 mg/kg. The attenuation of phase shifts by CP154,526 was specific to phase advances as light-induced phase delays of the circadian pacemaker achieved early in the subjective night were not affected by CP154,526 (20 mg/kg). We also tested one of the CRF(1) receptor antagonists for its potential ability to reset the pacemaker in the absence of light and found that CP154,526 did not elicit a nonphotic phase shifts in circadian activity rhythms at circadian times (CT) 2, 8, 14, 18, or 22. In conclusion, CRF(1) receptor antagonists selectively modulate the effect of light on the circadian pacemaker late at night. These novel data emphasize the suspected critical link between CRF and the hypothalamic-pituitary-adrenal axis, on the one hand, and stress (including stress caused by jet-lag) and depression on the other. These results also suggest that CRF(1) antagonists may not only improve affect but also counter the circadian disruption associated with depression and other stress-related disorders.

The selective tachykinin neurokinin 1 (NK1) receptor antagonist, GR 205,171, stereospecifically inhibits light-induced phase advances of hamster circadian activity rhythms.

Circadian rhythms in mammals are generated by master pacemaker cells located within the suprachiasmatic nucleus of the hypothalamus. In hamsters, the suprachiasmatic nucleus contains a small collection of cells immunoreactive for substance P, the endogenous ligand of tachykinin neurokinin 1 (NK1) receptors. In addition, two other nuclei which form part of the circadian system, the intergeniculate leaflet of the thalamus and the raphe nuclei, also contain fibers and/or cell bodies immunoreactive for substance P. In light of these observations, we evaluated the influence of the selective tachykinin NK1 receptor antagonist, GR 205,171, upon circadian activity rhythms in the hamster. Systemic injection of GR 205,171 dose-dependently (2.5-40.0 mg/kg, i.p.) inhibited light-induced phase advances in hamster circadian wheel running activity rhythms by approximately 50%. In contrast, GR 226,206, the less active enantiomer of GR 205,171, failed to affect light-induced phase advances. In addition, we examined the potential ability of GR 205,171 to induce non-photic phase shifts in hamster wheel running rhythms when injected at mid-day to late night circadian times. However, GR 205,171 (40 mg/kg) did not elicit non-photic phase shifts at these times indicating that tachykinin NK1 receptor antagonists are only effective when a light stimulus is applied to the pacemaker. Although GR 205,171 may, in theory, activate several sites within the circadian system, we suggest that GR 205,171 acts in the raphe nuclei to increase inhibitory serotonergic input to pacemaker cells in the suprachiasmatic nuclei, thereby suppressing photic modulation of the pacemaker. These findings have important implications for the use of tachykinin NK1 receptor antagonists in the treatment of depression and other central nervous system disorders.

Expression of the transcriptional coactivators CBP and p300 in the hamster suprachiasmatic nucleus: possible molecular components of the mammalian circadian clock.

Immediate early genes are expressed in the mammalian suprachiasmatic nucleus in response to photic information arriving from the retina at restricted times of the day, therefore their expression is regulated by the circadian biological clock. These light-induced genes are also activated by the phosphorylated form of CREB (pCREB) that binds to a cAMP response element upstream of the genes. The nuclear proteins CBP and p300 are known to be coactivators with pCREB in certain cell types, but their identification within the rodent SCN has not been reported. Therefore, in this study we examined the distribution of both CBP and p300 in the hamster suprachiasmatic nucleus. CBP and p300 immunoreactivity is detected in cells throughout the suprachiasmatic nucleus, and the pattern of staining within cells is indicative of a nuclear location for these proteins. The number of cells immunoreactive for both CBP and p300 significantly decreases at mid-night circadian times with respect to mid-day circadian times, although the reduction is less than 20%. Neither CBP nor p300 expression is affected by a circadian phase-resetting light pulse given late in the night. The ability of CBP and p300 to interact with pCREB as well as with the clock gene BMAL1 is discussed, and we propose that CBP and p300 may interact with, and link, both clock genes and clock-controlled genes in the generation of circadian rhythms in mammals. We further suggest that there will be a general importance for the role of transcriptional coactivators such as CBP and p300 in many of the molecular pathways related to the mammalian circadian clock.

Delta opioid inhibition of light-induced phase advances in hamster circadian activity rhythms.

A master neuronal pacemaker located within the suprachiasmatic nucleus in the ventral hypothalamus generates circadian activity rhythms in hamsters. The circadian pacemaker receives afferent input from many brain regions, one of which is the intergeniculate leaflet of the thalamus. This thalamic input to the suprachiasmatic nucleus in hamsters contains enkephalins, neuropeptide Y, neurotensin, and GABA. The role of enkephalins in modulating light-induced phase shifts of hamster activity rhythms has not been reported. Therefore, in this study, we examined the ability of enkephalin-mimetic and other opioid compounds to modulate light-induced phase advances in hamster circadian activity rhythms. The delta opioid agonists SNC 80 and BW373U86 both inhibited light-induced phase advances of hamster circadian activity rhythms. Neither the mu opioid agonist morphine, nor the kappa opioid agonist U50488H had any effect on light-induced phase shifts. The antagonists naltrindole, naltrexone, and nor-binaltorphimine, selective for delta, mu, and kappa opioids respectively, were also without effect on light-induced phase advances. Therefore, we found that only delta opioid agonists modulate light-induced phase advances in hamster circadian activity rhythms. These results imply that enkephalins released from the intergeniculate leaflet onto components of the suprachiasmatic pacemaker may be capable of inhibiting the responsiveness of the pacemaker to photic input arriving from the retina. The inability of antagonists to modulate light-induced phase advances suggests that endogenous opiate systems are not tonically active in generating circadian activity rhythms, but rather that enkephalins are probably used by the circadian system to modulate responses only under certain conditions or time of day.