Injections Of Triazolam Research Articles

The Non-Benzodiazepine Hypnotic Zolpidem Impairs Sleep-Dependent Cortical Plasticity

The effects of hypnotics on sleep-dependent brain plasticity are unknown. We have shown that sleep enhances a canonical model of in vivo cortical plasticity, known as ocular dominance plasticity (ODP). We investigated the effects of 3 different classes of hypnotics on ODP. Polysomnographic recordings were performed during the entire experiment (20 h). After a baseline sleep/wake recording (6 h), cats received 6 h of monocular deprivation (MD) followed by an i.p. injection of triazolam (1-10 mg/kg i.p.), zolpidem (10 mg/kg i.p.), ramelteon (0.1-1 mg/kg i.p.), or vehicle (DMSO i.p.). They were then allowed to sleep ad lib for 8 h, after which they were prepared for optical imaging of intrinsic cortical signals and single-unit electrophysiology. Basic neurophysiology laboratory Cats (male and female) in the critical period of visual development (postnatal days 28-41) N/A. Zolpidem reduced cortical plasticity by approximately 50% as assessed with optical imaging of intrinsic cortical signals. This was not due to abnormal sleep architecture because triazolam, which perturbed sleep architecture and sleep EEGs more profoundly than zolpidem, had no effect on plasticity. Ramelteon minimally altered sleep and had no effect on ODP. Our findings demonstrate that alterations in sleep architecture do not necessarily lead to impairments in sleep function. Conversely, hypnotics that produce more "physiological" sleep based on polysomnography may impair critical brain processes, depending on their pharmacology.

Behavioral and Serotonergic Regulation of Circadian Rhythms

Endogenous depression is often accompanied by alterations in core parameters of circadian rhythms, and antidepressant treatments, including serotonergic drugs, sleep deprivation and exercise, alter circadian phase or period in humans or animal models. Antidepressants may act in part through the circadian system, and behavioral antidepressants through a common serotonergic path to the clock. This review evaluates the evidence from animal models that serotonin (5-HT) mediates phase-shifting effects of behavioral stimuli on circadian rhythms. In rodents, 'exercise' stimulated during the rest phase of the rest-activity cycle induces large phase shifts of circadian rhythms. These shifts can be mimicked by short-term sleep deprivation without intense activity. During wheel running or sleep deprivation, 5-HT release in the suprachiasmatic nucleus (SCN) circadian clock is significantly elevated. Lesions of 5-HT afferents to the SCN attenuate phase shifts or entrainment induced by activity in response to some stimuli (e.g., triazolam injections in hamsters, treadmill running in mice) but not others (e.g., novel wheel confinement in hamsters). Antagonists selective to 5HT1, 2 or 7 receptors do not attenuate shifts induced by wheel running, although 5-HT2/7 antagonists do partially block shifts to saline injections. 5-HT agonists (e.g., 8-OH-DPAT) induce large shifts in vitro, but much smaller shifts in vivo, particularly if administered directly to the SCN. Procedures for inducing 5-HT supersensitivity in vivo result in larger shifts to 8-OH-DPAT. 5-HT stimuli may affect the clock by direct and indirect pathways, particularly through the thalamic intergeniculate leaflet, and the role of these pathways may differ across species. At the level of the SCN, 5-HT likely acts through 5-HT7 receptors on neurons and possibly also glial cells. These receptors may be useful targets for the development of antidepressant drugs. In aggregate, the literature provides mixed support for the hypothesis that exercise or behavioral arousal shift the circadian clock by a 5-HT pathway; the role of indirect pathways, interactions with other transmitters, cellular adaptations to denervation, glial cells, and species differences remain to be more fully clarified. Serotonergic and behavioral stimuli provide an intriguing route to elucidate the circadian clockworks and their possible role in depression.

Daily exposure to a nonphotic stimulus can alter photoperiodic response to short days in hamsters.

The ability of mammals to measure seasonal changes in daylength depends upon a circadian clock and the phase-relationship between this clock and the light: dark cycle. Recently, a number of pharmacological and nonpharmacological stimuli have been shown to have pronounced effects on the phase of the circadian clock of rodents. The objective of the present study was to determine if a drug-induced change in the phase-relationship between a measurable circadian rhythm (i.e., wheel running behavior) and the light:dark cycle would alter the effects of the light cycle on the neuroendocrine-gonadal axis. Adult male hamsters with regressed testes due to exposure to an inhibitory 10:14-hr light:dark cycle were daily injected with vehicle or the short-acting benzodiazepine, triazolam, while remaining on short days, while a control group of hamsters was transferred to a photostimulatory 14:10-hr light:dark cycle. Two other groups of hamsters with regressed testes were blinded and daily injected with vehicle or triazolam. The injections were timed to occur about 4 hr before activity onset because previous studies had demonstrated that injections of triazolam at this time can lead to a phase advance in the activity rhythm. The circadian rhythm of wheel running behavior was measured in all the animals maintained on the 10:14-hr light:dark cycle in order to monitor circadian phase. While no testicular growth was observed after 25 days of vehicle injections, growth was observed in the triazolam-treated animals that was comparable to that observed in control animals transferred to long days. Testicular growth in triazolam-treated animals was associated with an earlier onset of locomotor activity, when compared with the vehicle-treated animals. Importantly, triazolam had no effect on the testicular size of blind animals. These results indicate that daily injections of triazolam can stimulate neuroendocrine-gonadal activity by altering the phase-relationship between the cycle and the circadian clock involved in photoperiodic time measurement, and that agents which can affect the clock may be useful in altering seasonal cycles.

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

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

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.

Neuropeptide Y microinjected into the suprachiasmatic region phase shifts circadian rhythms in constant darkness

The geniculohypothalamic tract (GHT) is a projection from the intergeniculate leaflet to the suprachiasmatic nucleus (SCN). The GHT exhibits neuropeptide Y (NPY) immunoreactivity and appears to communicate photic information to the SCN. Microinjection of NPY into the SCN has been found to phase shift circadian rhythms of hamsters housed in constant light in a manner similar to the phase shifts produced by pulses of darkness or triazolam injections. In the present study, NPY was injected into the SCN of Syrian hamsters housed in constant darkness and was found to produce phase shifts similar to those seen in hamsters housed in constant light. Microinjections were not followed by wheel running during the subjective day (the time when NPY microinjections are followed by significant phase advances). These data suggest that NPY produces phase shifts by some mechanism other than by inducing wheel running or by inhibiting the response of SCN neurons to light and supports a role for NPY in nonphotic shifting of the circadian clock.

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 ☐es 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.

Aging alters the entraining effects of an activity-inducing stimulus on the circadian clock

In young hamsters, a single injection of the short-acting benzodiazepine, triazolan, can induce permanent phase shift in the circadian clock, while repeated injections of triazolam entrain the circadian clock to the period of the injections. Triazolam appears to act on the circadian clock by inducing an acute increase in the activity of the animals, which in turn phase-shifts the circadian clock. Surprisingly, single injections of benzodiazepines do not phase-shift the activity rhhthm of old hamsters, despite the fact that such treatment induces acute changes in the activity state young and old animals. We compared the entraining effects of repeated injections of triazolam on the circadian clock of young and old hamsters: while six out of seven young hamsters were entrained to the triazolam injections, only one out of seven old animals was entrained by this treatment. Three of the remaining six old hamsters showed a lengthening of the activity rhythm, while no consistent effect on the period of the activity rhythm was observed in the remaining three old animals. These results indicate that the circadian system of old hamsters becomes selectively unresponsive to synchronizing signals mediated by the activity-rest state, and suggest that aging is associated with a weakened coupling between the activity-rest cycle and the circadian clock.

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.

Daily injections of triazolam induce long-term changes in hamster circadian period

Previous experiments in hamsters indicate that daily injections of the short-acting benzodiazepine, triazolam, can entrain the free-running circadian activity rhythm to the period of the injections and that, after discontinuation of triazolam injections, the period of the free-running activity rhythm remains close to that of the previous injection schedule for 20-50 cycles. In this paper, we extend these findings and demonstrate that 1) long-term treatment with triazolam can induce aftereffects on the period of the circadian clock underlying the activity rhythm that can last for up to 100 days, 2) triazolam-induced changes in period can lead to a loss of effect of daily injections of triazolam which can be restored if the time of injection is altered, and 3) chronic treatment with triazolam also alters the period of the circadian clock in animals entrained to a light-dark cycle, and such changes in period alter the phase relationship between the circadian clock and the entraining light-dark cycle.

Stimulated activity mediates phase shifts in the hamster circadian clock induced by dark pulses or benzodiazepines

A number of environmental and pharmacological stimuli capable of inducing phase shifts and/or period changes in the circadian clock of mammals have now been identified. Agents that can alter circadian clocks provide a means for investigating the cellular and neural mechanisms responsible for their generation, regulation and entrainment. Two stimuli that have been used to probe the basis of circadian rhythmicity are pulses of darkness on a background of constant light and injections of short-acting benzodiazepines, such as triazolam. Surprisingly, these two very different stimuli have remarkably similar phase-shifting effects on the circadian clock of hamsters. The observation that a short-term increase in locomotor activity occurs when the circadian activity rhythm of hamsters is shifted by dark pulses or triazolam injections, coupled with the finding that activity bouts themselves are capable of shifting this rhythm, raises the possibility that dark pulses or triazolam alter the circadian clock by inducing acute hyperactivity. Here we demonstrate that the phase-advancing and phase-delaying effects of dark pulses or triazolam on the circadian activity rhythm can be totally suppressed by immobilization of the animals during treatment. These results indicate that behavioural events mediate the phase-shifting effects of both dark pulses and triazolam on the circadian activity rhythm and question present hypotheses regarding the pathways by which light-dark information and pharmacological agents influence circadian pacemakers.

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)

Single injections of triazolam, a short-acting benzodiazepine, lengthen the period of the circadian activity rhythm in golden hamsters.

Single injections of the benzodiazepine, triazolam, induce phase shifts and cause a lengthening of the circadian activity rhythm in the golden hamster. The effect of triazolam on period depends on the phase of injection, but is not dependent on the direction of the phase shifts. Triazolam injections caused increases in period that were associated with phase advances as well as phase delays in the activity rhythm. This relationship between triazolam-induced phase shifts and changes in period is different from the relationship between light-induced phase shifts and period changes.

Use of benzodiazepines to manipulate the circadian clock regulating behavioral and endocrine rhythms.

Extensive studies have now been carried out demonstrating that the systemic administration of the short-acting benzodiazepine, triazolam, can have pronounced effects on both behavioral and endocrine circadian rhythms. For example, three daily injections of triazolam can phase-advance the circadian rhythm of pituitary luteinizing hormone release and locomotor activity by about 2-3 h in female hamsters maintained in constant light. Triazolam has also been found to facilitate the rate of reentrainment of the activity rhythm following an 8-hour advance or delay in the light-dark cycle. Limited studies with other short-acting benzodiazepines indicate that the effects of triazolam on the circadian system of hamsters can be generalized to this class of drugs. Recent studies in humans indicate that treatment with triazolam can alter the time it takes for human endocrine rhythms to become reentrained following an 8-hour delay in the sleep-wake and light-dark cycle. Such findings raise the possibility that short-acting benzodiazepines may 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 of biological timekeeping.

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.

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.

Arousal induced by injection of triazolam into the dorsal raphe nucleus of rats.

In order to explore the possible sites at which benzodiazepines alter sleep, we have performed sleep studies following administration of 0.5 microgram of triazolam into the dorsal raphe nucleus of rats. Triazolam significantly increased sleep latency and decreased non-rapid eye movement (REM) sleep, with an effect greatest in the first 2 hours after injection. Total REM sleep time was not significantly affected, although there was a modest trend toward reduction in the first 2 hours. In contrast to the decreased sleep resulting from injection into the dorsal raphe nucleus, triazolam did not significantly alter sleep in animals in whom it was injected into surrounding areas. Similarly, the low dose employed in this study did not significantly affect sleep when injected into the lateral ventricle. These data are reminiscent of studies showing transient decreases in sleep following lesions of the dorsal raphe nucleus.

Entrainment of the circadian activity rhythm to the light-dark cycle can be altered by a short-acting benzodiazepine, triazolam.

The circadian rhythm of locomotor activity in hamsters maintained in either constant darkness or constant light can be phase-shifted by a single injection of the short-acting benzodiazepine, triazolam. These results suggest that treatment with triazolam may also alter the entrainment pattern of circadian rhythms in animals that are synchronized to a light-dark (LD) cycle. To test this hypothesis, hamsters maintained on an LD 6:18 light cycle received daily injections of triazolam (or vehicle) for 10-12 days, and any subsequent effects on the phase relationship between the onset of activity and the LD cycle were determined. Daily injections of triazolam (but not vehicle) induced pronounced advances or delays in the phase relationship between the entrained activity rhythm and the LD cycle; the direction of the shift was dependent on the time of the injection. Taken together with data from previous studies, these results suggest that triazolam, and perhaps other short-acting benzodiazepines, can be used to manipulate the mammalian circadian clock under a variety of experimental conditions.

Dose response curve for the phase-shifting effect of triazolam on the mammalian circadian clock

A dose response curve for the phase shifting effect of triazolam, a short-acting benzodiazepine commonly prescribed for the treatment of insomnia, on the circadian rhythm of locomotor activity was measured for the golden hamster. A single intraperitoneal injection of triazolam six hours before the onset of wheel-running activity induced a dose-dependent phase advance in the rhythm. A maximum phase advance, which averaged about 100 minutes, was observed in animals injected with 0.5 to 5.0 mg of triazolam. The use of drugs which promote sleep, and induce phase shifts in a central circadian clock, could be important in the treatment of sleep disorders associated with disrupted schedules and of mental and physical disorders associated with abnormal circadian rhythmicity.