Portacaval anastomosis disrupts circadian locomotor activity and pineal melatonin rhythms in rats

To determine whether hepatic encephalopathy may be associated with a disruption of circadian function, the circadian rhythms of locomotor activity and pineal melatonin content were examined in an animal model of complete portal-systemic shunting, rats with a portacaval anastomosis (PCA). The locomotor activity rhythm of all sham-operated animals entrained normally to a light/dark cycle and exhibited a normal free-running period during exposure to constant light. In contrast, PCA led to a dampening of the locomotor activity rhythm in all animals and the abolishment of a circadian periodicity in the activity rhythm of approximately 50% of rats during exposure to either a light/dark cycle or constant light. While normal diurnal variations of pineal melatonin content were seen in sham-operated rats, the amplitude of this variation appeared to be decreased in PCA animals. The similar effects of PCA on both a behavioral and an endocrine circadian rhythm, known to be regulated by a common neural pacemaker, coupled with studies indicating that a variety of other circadian rhythms may be disrupted in both animals and humans with hepatic dysfunction, suggests that this circadian disturbance originates within the pacemaker or on one of its afferent/efferent pathways.     

Clock genes outside the suprachiasmatic nucleus involved in manifestation of locomotor activity rhythm in rats

Chronic treatment of methamphetamine (MAP) in rats desynchronized the locomotor activity rhythm from the light-dark cycle. When the activity rhythm was completely phase-reversed with respect to a light dark-cycle, 24 h profiles were examined for the clock gene (rPer1, rPer2, rBMAL1, rClock) expressions in several brain structures by in situ hybridization, and for the pineal as well as plasma melatonin levels. In the MAP-treated rats, the rPer1 expression in the suprachiasmatic nucleus (SCN) showed a robust circadian rhythm which was essentially identical to that in the control rats. Circadian rhythms in pineal as well as plasma melatonin were not changed significantly in the MAP-treated rats. However, robust circadian rhythms in the rPer1, rPer2 and rBMAL1 expressions detected in the caudate-putamen (CPU) and parietal cortex were completely phase-reversed in the MAP-treated rats, compared with those in the control rats, indicating desynchronization from the SCN rhythm. Such desynchronization was not observed in the circadian rhythms of clock gene expression in the nucleus accumbens and cingulate cortex. The circadian rClock expression rhythm in the MAP-treated rats was not phase-reversed in the CPU and parietal cortex. These findings indicate that the locomotor activity rhythm in rats is directly driven by the pacemaker outside the SCN, in which rPer1, rPer2 and rBMAL1 in the CPU and parietal cortex are involved.     

Circadian rhythm of drinking and running-wheel activity in rats with 6-hydroxydopamine lesions of the ventral tegmental area

Circadian rhythms in drinking and running-wheel (locomotor) activity of rats with 6-hydroxydopamine (6-OHDA, 4 microg/2 microl per rat)-induced lesions in the ventral tegmental area (VTA) were examined under a light-dark (LD) cycle and constant dim light (5 lux). Under the LD cycle, the length of the locomotor activity period was decreased during the dark, and increased during the light period in the lesioned rats. Under the constant dim light conditions, the free-running circadian period (tau) of drinking and activity rhythm was longer in lesioned rats than in sham-operated controls. The elongation of the circadian period was accompanied by decrements in activity. These observations suggest that the mesolimbic dopaminergic system modulates rhythms in circadian drinking and locomotor activity.     

Ultradian and circadian body temperature and activity rhythms in chronic MPTP treated monkeys.

The body temperature and locomotor activity rhythms of seven 1-Methyl, 4-phenyl, 1,2,3,6-tetrahydropyridine (MPTP)-treated cynomolgous monkeys were registered over a week on two separate occasions over an interval of 2 months. Motor disability was absent in two animals and present in five: it was mild in one, moderate in two and severe in two. Both temperature and motor activity were recorded every minute using a radio telemetry system. Analysis of circadian rhythms revealed less robustness of the 24-hour circadian components of body temperature and locomotor activity with increasing motor impairment, and a fragmentation of the body temperature rhythm into 8 hour-period components. Both total activity and daytime activity correlated inversely with the degree of motor impairment. On the contrary, the monkeys did not show differences in night time activity. The proportions of variance accounted for by the body temperature and locomotor activity of 24 h + 12 h + 8 h components were correlated. Also, the average levels at which the circadian rhythm varies between body temperature and locomotor activity were correlated. The results were almost identical in the two 1-week recording sessions. The present study confirms individual differences in the vulnerability to MPTP of the nigrostriatal system of monkeys, suggesting that if a cumulative dose does not provoke stable motor alterations, this cumulative dose will not produce circadian body temperature and locomotor activity rhythm alterations either. Similarly, if a dose is able to produce motor impairment, this dose will also be able to produce circadian rhythm alterations.     

Circadian rhythm of nociception in the golden hamster

The response latency of golden hamsters to nociceptive stimuli was measured under cyclic lighting conditions and during constant illumination. A day-night rhythm of nociception was demonstrated; response latencies were significantly longer during the day. A circadian rhythm of nociception was displayed by hamsters maintained for 30 days in constant dim light. Short response latencies noted under these conditions were associated with the inactive period of the animals circadian cycle (subjective day). The experiments provide data which indicate the phase relationship between the circadian rhythms of nociception and locomotor activity differs under entrained and free-running conditions.     

Circadian rhythm of Arg–vasopressin contents in the suprachiasmatic nucleus in relation to corticosterone

Circadian rhythms of locomotor activity and adrenal glucocorticoid are controlled by the suprachiasmatic nucleus (SCN), the center of a biological clock, in mammals. Arg–vasopressin (AVP) contents in the SCN play a role in endogenous circadian rhythm during the absence of time cues. The AVP-containing neurons in the SCN are considered to transmit a circadian signal to the other parts of the brain. The circadian rhythms of AVP in the SCN in relation to the plasma corticosterone and locomotor activity were investigated. Under the light–dark cycle, plasma corticosterone levels were reciprocally correlated with the AVP content in the SCN. Under free-running conditions with constant dim light, AVP rhythms were reciprocally synchronized with the locomotor activity. The correlation of AVP with plasma corticosterone is different at different times of the day both under the LD cycle and constant dim light. Dexamethasone (i.p., 0.1 mg/100) increased the AVP contents, and this tendency was significantly greater during the dark period. These results indicate that corticosterone in the blood may regulate the circadian rhythm through AVP variation in the SCN.     

MKC-231, a choline uptake enhancer: (3) mode of action of MKC-231 in the enhancement of high-affinity choline uptake

MKC-231, a putative cholinergic activity, is reported to improve learning and memory impaired in AF64A-treated animals. MKC-231 enhances high-affinity choline uptake (HACU) known as the rate-limiting step of acetylcholine (ACh) synthesis. We investigated the mode of action (MOA) of HACU enhancement by MKC-231. Intracerebroventricular (i.c.v.) injections of AF64A (3 nmol/brain) resulted in significant HACU reduction in hippocampal synaptosomes. Treatment with MKC-231 increased Vmax of HACU and Bmax of [3H]-HC-3 binding 1.6 and 1.7-fold, respectively. In studies of [3H]-MKC-231 binding and Biacore analysis, MKC-231 showed noticeable affinity for cloned high-affinity choline transporters (CHT1). The present study suggests that MKC-231 directly affects trafficking of CHT1 and increases the numbers of transporter, working for HACU, at the synaptic membrane.     

Effects of methamphetamine on development of circadian rhythms in rats

To determine the effect of continuous methamphetamine treatment on development of the circadian system, the daily courses of spontaneous locomotor activity and plasma corticosterone level were determined in infant rats. Methamphetamine was dissolved in drinking water at a concentration of 0.005%, and administered to rats from postnatal day 14. The circadian locomotor rhythm in methamphetamine treated rats was not significantly different from that in non-treated rats for the first 4 weeks of drug treatment. Then the rhythm began to show signs of relative coordination and finally split into two activity components. One component entrained to the light-dark cycle, whereas the other free-ran in spite of the light cycle. The rhythm of plasma corticosterone in the methamphetamine treated group was not different from that in the control group at 4 weeks after birth, but was significantly phase-delayed at 8 weeks. Splitting was not observed for the corticosterone rhythm. It is concluded that continuous administration of methamphetamine to infant rats affects the circadian rhythms for spontaneous locomotor activity and plasma corticosterone level. Possible mechanisms of methamphetamine effects were discussed in terms of the circadian system underlying these rhythms.     

Temporal profile of circadian clock gene expression in a transplanted suprachiasmatic nucleus and peripheral tissues

The mammalian hypothalamic suprachiasmatic nucleus (SCN) is the master oscillator that regulates the circadian rhythms of the peripheral oscillators. Previous studies have demonstrated that the transplantation of embryonic SCN tissues into SCN-lesioned arrhythmic mice restores the behavioral circadian rhythms of these animals. In our present study, we examined the clock gene expression profiles in a transplanted SCN and peripheral tissues, and also analysed the circadian rhythm of the locomotor activity in SCN-grafted mice. These experiments were undertaken to elucidate whether the transplanted SCN generates a dynamic circadian oscillation and maintains the phase relationships that can be detected in intact mice. The grafted SCN indeed showed dynamic circadian expression rhythms of clock genes such as mPeriod1 (mPer1) and mPeriod2 (mPer2). Furthermore, the phase differences between the expression rhythms of these genes in the grafted SCN and the locomotor activity rhythms of the transplanted animals were found to be very similar to those in intact animals. Moreover, in the liver, kidney and skeletal muscles of the transplanted animals, the phase angles between the circadian rhythm of the grafted SCN and that of the peripheral tissues were maintained as in intact animals. However, in the SCN-grafted animals, the amplitudes of the mPer1 and mPer2 rhythms were attenuated in the peripheral tissues. Our current findings therefore indicate that a transplanted SCN has the capacity to generate a dynamic intrinsic circadian oscillation, and can also lock the normal phase angles among the SCN, locomotor activity and peripheral oscillators in a similar manner as in intact control animals.     

Behavioural rhythm splitting in the CS mouse is related to clock gene expression outside the suprachiasmatic nucleus

CS mice exhibit a spontaneous splitting in the circadian rhythm of locomotor activity under constant darkness, suggesting that they contain two weakly coupled oscillators in the circadian clock system regulating locomotor activity rhythm. In order to clarify whether the two oscillators are located in the suprachiasmatic nucleus (SCN), a site of the master circadian pacemaker in mammals, circadian rhythms in mRNA of mouse Period genes (mPer1, mPer2 and mPer3) in the SCN and cerebral cortex were examined during rhythm splitting by in situ hybridization. In the SCN, mPer1 and mPer2 showed a circadian rhythm with a single peak in both split and unsplit mice. The rhythms of mPer1 and mPer2 were slightly phase delayed during rhythm splitting in reference to the activity onset, but the phase relationship between the two rhythms was not changed. In the cerebral cortex, the expression of mPer1 and mPer2 underwent the bimodal fluctuation with peaks temporally corresponding to split activity components. The unsplit mice showed the circadian rhythms with a single peak. There was no difference in the mPer3 rhythms in either the SCN or the cerebral cortex between the split and unsplit mice. These results indicate that the circadian oscillations of mPer1, mPer2 and mPer3 in the SCN are not related to the rhythm splitting of CS mice. The split rhythms of the CS mice are suggested to be caused by uncoupling of oscillators located outside the SCN from the SCN circadian pacemaker.     

Circadian rhythms, aging and memory

In human beings and animal models, cognitive performance is often impaired in natural and experimental situations where circadian rhythms are disrupted. This includes a general decline in cognitive ability and fragmentation of behavioural rhythms in the aging population of numerous species. There is some evidence that rhythm disruption may lead directly to cognitive impairment; however, this causal link has not been made for effects due to aging. We have tested this link by examining rhythms and performance on contextual conditioning with the conditioned place preference task, in elderly, age-matched hamsters. Young healthy hamsters developed a preference for a context that is paired with the opportunity to engage in wheel-running (experiment 1). Aged animals with consolidated locomotor rhythms developed similar degrees of preference, whereas the age-matched hamsters with fragmented rhythms did not (experiment 2). The degree of preference was also correlated with activity amplitude. These results support the notion that age-related rhythm fragmentation contributes to the age-related memory decline.     

Light transducer for the biological clock: A function for rapid eye movements

Summary To test the hypothesis that the rapid eye movements (REMs) of paradoxical sleep (PS) transmit information about environmental light: dark cycles, spontaneous locomotor patterns were studied in rats following surgical disconnection of the extraocular muscles and thus deprived only of the eye movement component of REM sleep. Entrainment of the circadian spontaneous activity cycle to the light: dark cycle was abolished after sectioning the extraocular muscles of rats, but not in sham-operated controls. Endogenous (free running) circadian patterns of spontaneous activity were preserved in the operated animals. The results are consistent with our electrophysiologic studies showing that REMs induce light modulated electrical potentials in brain regions that influence circadian rhythms of activity.     

Circadian rhythmicity restored by neural transplant. Immunocytochemical characterization of the graft and its integration with the host brain

It is well established that overt circadian rhythms are permanently disrupted following lesions of the hamster hypothalamic suprachiasmatic nucleus (SCN). In the present study, we show that implantations of brain grafts containing the fetal SCN reestablish circadian rhythms of locomotor activity in adult hamsters previously made arrhythmic by SCN lesions. The restoration of free-running rhythms in conditions of constant darkness is correlated with the presence in the graft of neuropeptides normally present in the SCN of unlesioned hamsters, including vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), and vasopressin (VP). In several recipients, grafts were found to receive retinal input, and appeared to send efferents into the host brain. Not all functions of the SCN were reinstated by the graft: animals with restored locomotor rhythms did not show gonadal regression in the absence of light, and failed to synchronize (entrain) to light intensities to which SCN-intact animals responded.     

Neonatal exposure to constant light prevents anhedonia-like behavior induced by constant light exposure in adulthood

Depressive episodes are associated with disturbances in circadian rhythms, and constant illumination has been reported to induce depressive-like behavior in rodents. Rats kept in constant darkness express the endogenous circadian rhythm, and most animals under constant light conditions lose circadian locomotor rhythmicity. Exposure to constant light in rats during lactation was reported to prevent this loss of circadian rhythm in adulthood. Thus, the aim of the present study was to verify whether exposure to constant light during lactation prevents anhedonia-like behavior induced by constant light in adult rats. In experiment 1, we replicated the anhedonia-like effects of constant light in adult male rats. We showed that this effect is reversed by imipramine treatment in the drinking water. In experiment 2, we subjected rats to constant darkness (neonatal-DD), constant light (neonatal-LL) or to normal light/dark cycle (neonatal-LD) during the neonatal phase and evaluated them after constant light exposure in adulthood. The group exposed to constant light during the neonatal phase did not reduce their sucrose preference and exhibited greater locomotor activity than the other groups. The neonatal-DD group exhibited decreased sucrose preference earlier than controls and had higher serum corticosterone concentrations. Prevention of arrhythymicity might protect neonatal-LL rats from anhedonia-like behavior induced by constant light, whereas constant darkness during the neonatal phase rendered the neonatal-DD group more susceptible to depressive-like behavior. These results corroborate with the literature data indicating that circadian disruption may contribute in mood disorders and that early life stress can influence stress responsivity in adulthood.     

Effects of hypothyroidism on rat circadian activity and temperature rhythms and their response to light.

Male rats made hypothyroid by administration of propylthiouracil plus sodium ipodate in drinking water were compared to controls in terms of period of circadian activity and temperature rhythms, amount of gross motor activity, and mean temperature. Animals were studied under entrainment, constant darkness (DD), and constant dim light (LL). There was no difference in the period of the circadian activity rhythm between groups in DD. However, hypothyroid rats showed significant blunting of the period-lengthening response to increasing ambient illumination. As expected, the period of the circadian temperature rhythm increased in controls with increasing ambient illumination. In contrast, the period of the circadian temperature rhythm in hypothyroid animals actually shortened under LL compared to DD. This blunting of the period-lengthening response to increasing ambient illumination of both activity and temperature rhythms in hypothyroid animals could not be explained by differences in activity level or mean temperature between the groups.     

Manipulation of the circadian clock with benzodiazepines: implications for altering the sleep-wake cycle

Abnormal circadian rhythms have been linked to at least some forms of depression and to disturbances in the sleep-wake cycle. In addition, mental and physical disorders associated with rapid travel across time zones (i.e. the jet-lag syndrome) and with rotating shift-work schedules, are thought to involve a disruption of normal circadian rhythmicity. It might be possible to alleviate some of the adverse effects associated with abnormal circadian rhythms if pharmacological agents could be used to manipulate the central circadian pacemaker(s) that regulates these rhythms. Recent findings indicate that treatment with a short-acting benzodiazepine, triazolam, can induce major shifts in the circadian clock of golden hamsters. In the absence of a synchronizing light-dark cycle (i.e. during exposure to constant light or constant dark), a single injection of triazolam can induce a permanent phase shift in the circadian rhythm in locomotor activity. In addition, following a shift in the light-dark cycle, a single injection of triazolam can facilitate the time it takes for the activity rhythm to be resynchronized to the new lighting schedule. Triazolam, or drugs with similar phase-shifting effects on the mammalian circadian system, might be useful in the treatment of various sleep and mental disorders that have been associated with a disorder in circadian time-keeping in humans.     

MKC-231, a choline uptake enhancer: (2) Effect on synthesis and release of acetylcholine in AF64A-treated rats

The effect of MKC-231 on acetylcholine (ACh) synthesis and release was studied in the hippocampus of normal and AF64A-treated rats. AF64A (3 nmol/brain, i.c.v.) produced significant reduction of high-affinity choline uptake (HACU) and high K+-induced ACh release in hippocampal synaptosomes. Treatments with MKC-231 (10(-8) and 10(-7) M) showed significant reverse of the decrease in both HACU and ACh release. In hippocampal slices superfused with choline-containing artificial cerebro-spinal fluid (ACSF), high K+-induced ACh release was gradually decreased by repeated alteration of resting and high K+ stimulations in AF64A-treated rats. However, addition of MKC-231 (10(-8) to 10(-7) M) in the superfusate reduces this decrease. In vivo microdialysis studies indicate MKC-231 (10 mg/kg, p.o.) significantly reversed reduction of basal ACh concentrations in AF64A-treated rats, measured by radioimmunoassay without a cholinesterase inhibitor in the perfusate. These results indicate MKC-231 improves AF64A-induced cholinergic hypofunction by enhancing HACU, subsequently facilitating ACh synthesis and release in vitro and in vivo.     

Serotonergic modulation of the hamster wheelrunning rhythm: response to lighting conditions and food deprivation

The midbrain raphe complex innervates the circadian rhythm regulating system by direct projections to the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL). The present experiments examined the changes in circadian rhythm regulation consequent to the depletion of brain serotonin by central 5,7-dihydroxytryptamine (DHT) application. Adult male hamsters with access to running wheels were entrained to a light-dark cycle 14:10 (LD) of photoperiod, pre-treated with desmethylimipramine and given bilateral lateral ventricle infusions of 75 micrograms DHT/2.5 microliters 0.5% ascorbic acid in saline or vehicle only. Two separate experiments were performed. Four weeks after surgery, animals were transferred to either constant light (LL; Experiment 1) or constant dark (DD; Experiment 2). Animals remained in LL for 85 days, then were transferred to DD for 50 days, followed by a return to LD 14:10 for 14 days. Animals in Expt. 2 remained in DD for 55 days, were given 3 days food deprivation, then, beginning 35 days later, were periodically exposed to 30 min light pulses as a phase response curve (PRC) to light was generated. DHT treatment induced rapid appearance of advanced activity onset, delayed offset and longer duration of the nocturnal activity phase. DHT animals in LL had circadian locomotor rhythms much longer than control animals (24.43 +/- 0.04 vs 24.19 +/- 0.05 h) and normal circadian rhythmicity was rapidly lost by DHT animals in LL. There was no effect of DHT on circadian period in DD, but the DHT treated animals in DD had a larger phase delay region of the PRC than did controls and this was associated with an overall change in the temporal properties of the PRC. Serotonin immunohistochemistry showed an approximate 90% loss of cells from the dorsal raphe nucleus and decreased density of the serotonergic terminal field in the SCN and IGL. The results support the view that the serotonergic system modulates the phasic actions of light on the hamster circadian rhythm system. The data also indicate that hamsters can have a Type 0 PRC.     

Altering the mammalian circadian clock with the short-acting benzodiazepine, triazolam

The systemic administration of triazolam alters circadian rhythms of hamsters under both entrained (synchronized to a circadian period) and free-running (in the absence of a synchronizing light-dark cycle) conditions. In the latter conditions (i.e. during exposure to constant light or darkness), single injections of triazolam can induce a permanent phase-shift in both behavioral and endocrine rhythms. Repeated injections at fixed circadian intervals can entrain the rhythm of locomotor activity, and in the presence of a synchronizing light—dark cycle, daily injections of triazolam can alter the phase relationship of the activity rhythm to the light—dark cycle. Moreover, following a shift in the light—dark cycle, a single injection of triazolam can shorten the time it takes for the activity rhythm to be resynchronized to the new lighting schedule. In addition to implicating a role for the neurotransmitter GABA in the circadian organization of mammals, these findings may have important clinical implications. If short-acting benzodiazepines can have similar effects on the human circadian clock, they could prove useful in reducing the symptoms associated with ‘jet-lag’ and rotating shift-work schedules, as well as in the treatment of various physical and mental illnesses that have been associated with a disorder in biological timekeeping.     

Circadian rhythms, sleep, and the menstrual cycle

Women with ovulatory menstrual cycles have a circadian rhythm superimposed on the menstrual-associated rhythm; in turn, menstrual events affect the circadian rhythm. In this paper, we review circadian rhythms in temperature, selected hormone profiles, and sleep-wake behavior in healthy women at different phases of the menstrual cycle. The effects on menstrual cycle rhythmicity of disrupted circadian rhythms, for example, with shiftwork and altered circadian rhythms in women with menstrual-related mood disturbances, are discussed. Compared to the follicular phase, in the post-ovulation luteal phase, body temperature is elevated, but the amplitude of the temperature rhythm is reduced. Evidence indicates that the amplitude of other rhythms, such as melatonin and cortisol, may also be blunted in the luteal phase. Subjective sleep quality is lowest around menses, but the timing and composition of sleep remains relatively stable across the menstrual cycle in healthy women, apart from an increase in spindle frequency activity and a minor decrease in rapid eye movement (REM) sleep during the luteal phase. Disruption of circadian rhythms is associated with disturbances in menstrual function. Female shiftworkers compared to non-shiftworkers are more likely to report menstrual irregularity and longer menstrual cycles. There also is accumulating evidence that circadian disruption increases the risk of breast cancer in women, possibly due to altered light exposure and reduced melatonin secretion. Further investigations into the biological consequences of circadian disruption in women will offer insight into some menstrual-associated disorders, including mood changes, as well as reproductive function and possible links with breast cancer.