Rhythms In Hamsters Research Articles

Effects of Microinjections of Substance P Into the Suprachiasmatic Nucleus Region on Hamster Wheel-Running Rhythms

The suprachiasmatic nucleus (SCN) receives a direct retinal projection, which in rats includes substance P (SP)-immunoreactive retinal ganglion cells. While SP has been shown to have neurophysiological effects on SCN cells in Syrian hamsters and rats, it is not known what effects SP in the SCN has on circadian rhythms in hamsters. We examined this question using male Syrian hamsters that were implanted with cannulas aimed at the SCN region and maintained in constant dim red lighting conditions. Hamsters received 0.5 μl microinjections of saline or SP (500 pmol in saline) at a variety of circadian times (CT). Saline injections had little or no phase-shifting effects at any phase tested. SP had no significant effects at CT4–8, 16–20, or 20–24, but did cause small phase delays of −23.7 ± 7 min (mean ± sem) at CT12–16. In order to examine the dose-response relations of this effect, hamsters were also microinjected with 50 and 2500 pmol of SP at CT12–16. Both the 50 and 2500 pmol doses induced very small phase delays (−14.2 ± 7 min and −18.2 ± 5 min, respectively), indicating no obvious dose dependence within this range. These results do not suggest that SP alone in the SCN mimics light effects on circadian rhythms or is a key neurotransmitter involved in photic entrainment. It remains to be determined whether SP interacts with other transmitters in the SCN to modulate their effects on rhythm phase.

Thyrotropin-releasing hormone phase shifts circadian rhythms in hamsters.

The role of thyrotropin-releasing hormone (TRH) in regulating circadian rhythms was investigated by assessing the ability of TRH microinjections into the suprachiasmatic nucleus (SCN) to induce phase shifts in hamster wheel-running behavior. TRH injected into the SCN at 10 and 100 nM doses produced phase advances in wheel-running activity of 18.3 +/- 1.9 and 34.8 +/- 2.9 minutes, respectively, when administered at circadian time (CT) 6. Injections at CT 18 produced no effects. The temporal sensitivity of the SCN to TRH administration was examined by administering TRH at specific circadian times. TRH produced significant phase advances at CT 4, 6, and 8, while no significant changes in wheel-running onset were observed at other CT times. These studies represent the first evidence of TRH's ability to affect circadian function.

Nerve growth factor phase shifts circadian activity rhythms in Syrian hamsters

Nerve growth factor (NGF) receptors are found in high density in the rodent suprachiasmatic nucleus (SCN), a site which regulates mammalian circadian rhyrthms. We examined the effects of NGF (40 ng) or vehicle injections into the SCN, at circadian times (CT) 6, 14 or 22 on activity rhythms in hamsters maintained in constant darkness. NGF caused phase advances at CT6 (30.9 min) and Cf22 (36.9 min), and phase delays at CT14 (31.2 min). Saline and cytochrome-c administration had no phase-shifting effects at CT6 and CT22, but at CT14 cytochrome-c produced large phase delays, implying that NGF-induced delays at this phase may be non-specific. Similarities between NGF-induced shifts and those elicited by the cholinergic agonist carbachol suggest a common mode of action.

Twelve-hour phase shifts of hamster circadian rhythms elicited by voluntary wheel running.

Running in a novel wheel can serve as a nonphotic zeitgeber to entrain or phase shift circadian rhythms in hamsters. In this study, hamsters were entrained to a light:dark schedule of 14:10 h but had no access to running wheels. At four different phase points of the light cycle, hamsters were transferred to constant darkness and provided with running wheels. All hamsters began running shortly after transfer and were allowed to continue running at their own volition. Approximately 20% of the hamsters transferred at zeitgeber time (ZT) 23 (ZT 12 = lights out) ran more than 4 h after transfer and showed phase advances of the circadian activity rhythm by as much as 15 h, while hamsters that ran less than 4 h on average did not phase shift. A similar result was observed for hamsters transferred at ZT 2. Hamsters transferred at ZT 5 and 8 also did not phase shift if they ran less than 4 h, although the relation between longer runs and phase shifts became less evident. A sustained run in excess of 4 h appeared to be associated with large phase advances. These results show that under certain conditions, a single sustained bout of wheel-running activity is capable of phase shifting the circadian pacemaker by more than 12 h.

Analysis of the phase shifting effects of gastrin releasing peptide when microinjected into the suprachiasmatic region

The role of gastrin releasing peptide (GRP) in phase shifting circadian rhythms was investigated. Microinjection of GRP into the suprachiasmatic region (SCN) region produced only small changes in the phase of free-running circadian activity rhythms in hamsters housed in constant darkness. However, co-administration of GRP with vasoactive intestinal peptide (VIP) and peptide histidine isoleucine (PHI) produced significantly larger phase delays than GRP alone. These data support the hypothesis that GRP interacts with VIP and PHI within the SCN to induce phase shifts of circadian rhythms.

Suprachiasmatic nucleus lesions abolish and fetal grafts restore circadian gnawing rhythms in hamsters

It is widely accepted that the suprachiasmatic nuclei (SCN) of the hypothalamus serve as biological pacemakers, organizing daily activities. However some circadian rhythms are controlled by extra-SCN structures. Transplantation of fetal donor SCN in SCN-lesioned rodents induces recovery of rhythmic locomotor and drinking activities. Such grafts do not however, restore appropriate gonadal responses to photoperiodic stimuli. It is not known whether other behavioral rhythms are restored by fetal tissue grafts, or whether various responses are restored simultaneously. In the present study, we established that circadian rhythms of gnawing behavior are abolished following SCN lesions. Next, we measured both gnawing and wheel-running activity in SCN-lesioned hamsters following transplantation of fetal hypothalamic grafts containing the SCN. The results indicate that such grafts restore circadian rhythms of gnawing behavior, and that gnawing and wheel-running rhythms re-emerge at about the same time.

Tau changes after single nonphotic events.

Changes in the free-running period of the circadian rhythms of hamsters occur after single nonphotic events such as a 3-h pulse of running induced by being put in a novel wheel. These changes are mostly in the direction of longer periods, and can exceed 0.2 h; the magnitude of the effect depends on the circadian phase of the pulse. The phase response curves for period changes do not match up with those for phase shifts of the rhythms. Data on free-running rhythms after anisomycin injections and after novelty-induced wheel running in tau mutant hamsters support the view that period changes and phase shifts can occur independently of one another.

Ethanol and circadian rhythms in the syrian hamster: Effects on entrained phase, reentrainment rate, and period

Wheel-running rhythms were examined in male hamsters with access to 28% ethanol in lieu of water. One group was recorded in light-dark (LD) cycle that was phase advanced by 8 h on three occasions separated by 23–27 days. On two of the three occasions, hamsters were subjected to a 2- to 3-h cage change procedure designed to stimulate wheel running, which accelerates the rate of reentrainment to 8-h advances. Ethanol and control hamsters showed no group differences in rhythm amplitude, entrained phase, or reentrainment rate. Both groups showed faster reentrainment in the cage change condition. A second group of hamsters recorded in constant dim showed a small but significant lengthening of the free-running period of their wheel-running rhythm when provided with a 28% ethanol solution. Wheel running decreased during ethanol access in this group. Voluntary ethanol consumption evidently can slow the circadian pacemaker regulating activity rhythms in hamsters but has no measurable effect on photic entrainment or pacemaker response to LD shifts or nonphotic manipulations (stimulated activity). Period lengthening may be secondary to decreased activity, but other period-activity correlations obtained did not reveal a strong association between these two variables.

Phase response curve to anisomycin in tau mutant hamsters.

Administration of the protein synthesis inhibitor, anisomycin, to wild type hamsters produces phase shifts in their circadian rhythms that have similarities to shifts produced by non-photic behavioral stimulation. A mutation that shortens the period of rhythms in hamsters results in altered responsiveness to non-photic input. However, responses of the mutants to anisomycin are unaffected: their phase response curve (PRC) for anisomycin is similar to that of wild types. This suggests that 1) anisomycin is not acting on mechanisms specifically involved in non-photic behavioral phase shifting, and 2) the mutation affects the non-photic input pathway or the pacemaker itself at a point that is upstream from anisomycin's site of action.

Dietary calcium blocks lithium toxicity in hamsters without affecting circadian rhythms

Lithium can be toxic to rodents at plasma concentrations (0.6–1.2 mmol/L) that also phase delay circadian rhythms. In hamsters, raising the concentration of calcium in the diet from 0.1%–3% reduced weight loss and polydipsia caused by 0.4% lithium carbonate. Calcium ingestion did not affect plasma lithium concentration or the phase of the circadian wheel-running rhythm in lithium-treated animals. Calcium ingestion did not alter weight gain, salt or water intake, or circadian rhythms in hamsters not receiving lithium. Dietary calcium supplementation may prevent some toxic effects of lithium without interfering with other central nervous system actions.

Phase-shifting effects of acute increases in activity on circadian locomotor rhythms in hamsters

Experiments were conducted in golden hamsters to examine the relationship between induced acute increases in locomotor activity and phase shifts in the circadian clock underlying the rhythm of activity. Injections of the short-acting benzodiazepine triazolam (TZ) 6 h before the onset of activity resulted in an acute increase in activity and a phase advance in the rhythm of activity; injections of TZ induced larger phase shifts in animals housed without running wheels than in those housed with wheels. Transfer to a cage with access to a running wheel for 1 h at different circadian times induced large phase advances (mean of 2 h) and small phase delays depending on the circadian time of transfer. Maximal mean phase advances resulted when animals were transferred to a cage with wheel 3 h before activity onset, and at this circadian time there was a significant correlation between the magnitude of the phase shift and the amount of increase over baseline activity for the first hour after transfer. These results indicate that access to a running wheel in animals housed without wheels can be a significant phase-shifting stimulus to the circadian clock and that the phase shifts induced by injection of TZ or transfer to a new cage with wheel are related to the activity state of the animal or to the amount of locomotor activity that is induced at particular times.

The benzodiazepine triazolam phase-shifts circadian activity rhythms in a diurnal primate, the squirrel monkey ( Saimiri sciureus)

Triazolam can shift the phase of circadian rhythms in hamsters recorded in constant light or dark. This effect is apparently mediated by physical activity stimulated by the drug. We examined whether triazolam can shift the phase of circadian rhythms in a diurnal primate, the squirrel monkey, that is sedated by triazolam. Single injections of triazolam at 0.15–0.20 mg/kg induced phase advances or delays of monkey activity rhythms recorded in constant light. The phase-response curve is similar to that obtained for hamsters. Behavioral activation is evidently not necessary for a phase-shifting action of triazolam in this primate. A companion study in which triazolam was administered to squirrel monkeys in the dark in a light-dark cycle reentrainment paradigm failed to find evidence for a phase shifting action of triazolam. Shifts induced by triazolam in monkeys recorded in constant light may thus reflect changes in light exposure as a consequence of sedation or altered retinal processing of light.

Suprachiasmatic nuclei influence torpor and circadian temperature rhythms in hamsters.

Siberian hamsters were maintained in a short-day photoperiod (8 h light-day) at 15 degrees C; body temperature (Tb) and locomotor activity were telemetrically recorded at 10-min intervals over the course of 5 mo. Animals manifesting repeated torpor bouts (Tb less than 30 degrees C for several hours) were subjected to lesions of the suprachiasmatic nuclei (SCN), pinealectomy, or sham operations. In the 15 wk after surgery, none of the animals with bilateral lesions of the SCN exhibited torpor; circadian Tb and locomotor activity rhythms, as determined by cosinor and power spectral analysis, also were absent in SCN-lesioned hamsters. Pinealectomized animals and brain-lesioned hamsters with intact SCN had normal circadian temperature and activity rhythms and showed torpor for at least 4 wk postsurgically. Expression of torpor and circadian rhythms of Tb and activity are dependent on intact SCN and persist for several weeks in the absence of pineal secretory activity.

Midazolam, a short-acting benzodiazepine, resets the circadian clock of the hamster

Treatment with the short-acting benzodiazepine, triazolam, has been found to induce changes in both behavioral and endocrine circadian rhythms in hamsters. The objective of this study was to determine if these effects of triazolam could be generalized to other short-acting benzodiazepines. Therefore, the effects of midazolam on the biological clock of the hamster were examined in detail. A phase-response curve and a dose-response curve were measured to determine the effects of a single intraperitoneal injection of midazolam on the circadian clock of hamsters free-running in constant light. Midazolam injections produced maximal phase advances at circadian time (CT) 6 and 9 and maximal phase delays at CT 15 and 21. Doses of 2.5 mg or larger produced phase shifts that were significantly different from those produced by the vehicle controls. In addition, the phase-shifting effects of midazolam were completely blocked by administration of the benzodiazepine receptor antagonist, RO 15-1788, indicating that the phase-shifting actions of midazolam are mediated via benzodiazepine receptors. These results indicate that the previously reported effects of triazolam on the circadian clock can be generalized to other short-acting benzodiazepines.

Administering triazolam on a circadian basis entrains the activity rhythm of hamsters.

A single injection of the short-acting benzodiazepine, triazolam, can induce permanent phase shifts in the circadian rhythm of locomotor activity in free-running hamsters, with the direction and magnitude of the phase shifts being dependent on the circadian time of treatment. The shape of the "phase-response curve" to triazolam injections is totally different from that for light pulses. These findings raise the possibility that repeated injections of triazolam on a circadian basis might be capable of entraining the circadian pacemaker underlying the activity rhythm of hamsters and that the entrainment pattern might differ from that observed in animals entrained to light pulses. To test this hypothesis, blind hamsters received intraperitoneal injections of triazolam (or vehicle) every 23.34, 23.72, 24.00 or 24.66 h for 19-20 days, and the effect of these injections on the period of the rhythm of wheel-running behavior was determined during and after treatment. Repeated injections of 0.1 mg triazolam at these time intervals resulted in the entrainment of the activity rhythm in 36 of 40 animals, whereas 0 of 40 animals entrained to vehicle injections. Importantly, the phase relationship between triazolam injections and the circadian activity rhythm was dependent on the period of drug treatment and could be predicted from the phase-response curve to single injections of triazolam. These phase relationships are dramatically different from those observed between the activity rhythm and 1-h light pulses presented at similar circadian intervals.(ABSTRACT TRUNCATED AT 250 WORDS)

Masking of circadian activity rhythms in hamsters by darkness

Wheel-running activity was recorded in male golden hamsters (Mesocricetus auratus) kept in constant dim illumination. For periods of several weeks the lights in the cabinet were turned off daily at the same time of day, either for 1 h or 2 h. Despite these periodically recurring dark pulses, the circadian activity rhythms continued to free-run, and consequently crossed through the pulses at a more or less regular speed. During a dark pulse, the activity was usually enhanced. The amount of these masking effects varied with the phase of the circadian cycle at which the pulse occurred. The responses were maximal a few hours after the onset of spontaneous activity, and minimal during the rest-time of the animal.

Glutamate phase shifts circadian activity rhythms in hamsters.

The suprachiasmatic nuclei (SCN) have been identified as a pacemaker for many circadian rhythms in mammals. Photic entrainment of this pacemaker can be accomplished via the direct retino-hypothalamic tract (RHT). Glutamate is a putative transmitter of the RHT. In the present study it is demonstrated that glutamate injections in the SCN cause phase shifts of the circadian activity rhythm of the hamster. In contrast, glutamate injections outside the SCN or vehicle injections inside the SCN did not affect the circadian phase. These data suggest that glutamate could be involved in photic entrainment of the circadian pacemaker.

Manipulation of a central circadian clock regulating behavioral and endocrine rhythms with a short-acting benzodiazepine used in the treatment of insomnia

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 that are 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 of abnormal circadian rhythms if pharmacological agents could be used to manipulate the central circadian pacemaker(s) that regulate these rhythms. Studies in our laboratory indicate that treatment with a short-acting benzodiazepine, triazolam, can induce major shifts in both behavioral and endocrine circadian rhythms in hamsters under a variety of experimental conditions. In the absence of a synchronizing light-dark cycle (i.e. during exposure to constant light or constant dark), single or multiple injections of triazolam can induce a permanent phase shift in both the circadian rhythm of locomotor activity and the circadian rhythm of pituitary LH release. In addition, repeated daily injections of triazolam can alter the entrained phase relationship of the circadian activity rhythm to a fixed light-dark cycle, and 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. Thus, triazolam, or drugs with similar phase-shifting effects on the mammalian circadian system, might be useful in the treatment of various physical and mental illnesses that have been associated with a disorder in circadian time-keeping in humans.

Entrainment of split circadian activity rhythms in hamsters.

Hamsters that showed splitting of their circadian rhythms of wheel-running activity following long-term exposure to constant illumination (LL) were exposed to light-dark (LD) cycles with 2-hr dark segments, and with periods of 24.00, 24.23 or 24.72 hr. For comparison, hamsters showing nonsplit rhythms were also studied. In all cases of split rhythms, at least one of the two split components entrained to the LD cycles. In some animals, the second component continued to free-run until it merged with the entrained component, while in others, the second component also entrained to the LD cycle but maintained a stable phase angle of 6-14.5 hr relative to dark onset. These results were obtained in cases where the period of the LD cycle was shorter than that of the split rhythms and in cases where it was longer, implying that split components can be phase-advanced as well as phase-delayed by 2 hr of darkness. Three hamsters that showed stable entrainment of split rhythms were allowed to free-run in LL. The LD cycles were then reinstated, but instead of overlapping with the first component, as it did before, the dark segment was timed to overlap with the second. The entrainment patterns that ensued were similar to the ones obtained during the first LD exposure, indicating that the two split components respond to darkness in a qualitatively similar fashion. These results are further evidence that the pacemaker system underlying split circadian activity rhythms in hamsters is composed of two mutually coupled populations of oscillators that have similar properties, including a bidirectional phase response curve. Such a dual-oscillator organization may also underlie normal, or nonsplit, activity rhythms, as suggested by Pittendrigh and Daan (1976c), but the data are also compatible with the alternative view that the circadian pacemaker consists of a large number of coupled oscillators, which only dissociate into two separate populations in some animals under conditions of moderate LL intensity.

Adrenal corticoids in hamsters: role in circadian timing.

The 24-h patterns of circulating cortisol and corticosterone were determined in male hamsters housed under a 14:10 light-dark cycle. Corticoid levels varied significantly over the 24-h sampling period with peak levels of both hormones occurring near the onset of the daily dark phase. The ratio of cortisol to corticosterone changed dramatically during the day. Corticosterone levels were significantly higher than cortisol during the early part of the light phase; however, cortisol levels became significantly higher than corticosterone when both hormones began their daily rise. To examine whether the circadian rhythm of cortisol secretion could be involved in the physiological control of hamster circadian organization, cortisol was infused at approximately physiological levels into adrenalectomized hamsters either continuously or in a 24-h rhythm. No significant differences were observed in the timing of circadian wheel-running rhythms in hamsters housed in LD 16:8, LD 14:10, or LL when cortisol was infused continuously, in a 24-h rhythm that mimicked the cortisol rhythm of intact hamsters, or in a 24-h rhythm several hours out of phase with the rhythm of intact hamsters. Provision of cortisol in a 24-h rhythm appeared to promote the survival of adrenalectomized hamsters since hamsters receiving a 24-h pattern of cortisol survived the experimental protocol significantly longer than those receiving the same dose of cortisol continuously.