Biomedical subjects
Publications and source records attributed to F W Turek.
Melatonin is being touted as the latest cure-all. But in their eagerness to explain to the public the possible health benefits of this natural hormone, the mass media as well as some scientists have misrepresented the scientific data.
Several lines of evidence suggest the potential involvement of serotonergic pathways in mediating the effects of activity-inducing stimuli on the circadian clock in rodents. The aim of the present 3 experiments was to examine the effects of the serotonergic neurotoxin, p-chloroamphetamine (PCA, 10 mg/kg) on: (1) the monoamine levels of the hypothalamus, frontal cortex and hippocampus in the hamster; (2) the phase shifts in the circadian rhythm of locomotor activity of hamsters in response to treatment with the short-acting benzodiazepine, triazolam (7.5 mg/kg); and (3) the magnitude of the acute increase in locomotor activity associated with triazolam administration in this species. The administration of PCA to hamsters caused changes of specific monoaminergic systems in the hypothalamus, that were limited to a selective decrease in serotonin levels 7 days post-treatment. The phase shifts of the circadian clock in response to triazolam treatment at CT 6 were considerably attenuated following the administration of the 5-HT neurotoxin. The total amount and the profiles of triazolam-induced wheel-running and general cage activity between CT 6 and CT 12 were not significantly affected by the PCA treatment. The finding that a 5-HT neurotoxin can attenuate the phase-shifting effects of triazolam in hamsters, without interfering with its activity-inducing properties, suggests that serotonergic afferents might be involved in the mechanism for non-photic phase-shifting of the circadian system.
Rats were deprived of REM sleep for 24 h by keeping them on small platforms that were placed in a water bath (the platform method). Galanin coding mRNA was visualized using in situ hybridization, and cells expressing galanin mRNA were counted. In REM sleep-deprived animals the cell count was higher in the preoptic area and periventricular nucleus. Lesions of this area have been reported to induce wakefulness in cats and rats. Galanin administered into the lateral ventricle had no effect on sleep. We conclude that REM sleep deprivation can induce galanin gene expression in some brain areas, but galanin alone does not modify spontaneous sleep.
In addition to light, a variety of non-photic stimuli can induce phase shifts in the circadian clock of rodents. We have examined the effects of advanced age on the response of the circadian clock to both photic and non-photic stimuli in old hamsters (i.e., over 16 months of age). Among the age-related changes in the circadian rhythm of locomotor activity are: (1) alterations in the phase angle of entrainment to the light-dark cycle; (2) an altered response to the phase-shifting effects of light pulses; (3) changes in the time it takes to re-entrain to a new light-dark cycle; and (4) a loss of responsiveness to the phase-shifting or entraining effects of stimuli which induce an acute increase of activity. Many of the effects of ageing on the circadian clock system can be simulated in young animals by depleting brain monoamine levels, suggesting that ageing alters monoaminergic inputs to the clock. Some of the age-related changes in the response of the clock to an activity-inducing stimulus can be reversed by implanting old animals with fetal suprachiasmatic nuclear tissue. Determining the physiological basis of age-related changes in the responsiveness of the clock to both internal and external stimuli, and the mechanisms by which normal circadian functioning can be restored, should lead to new insight into the functioning of the circadian clock and may suggest new approaches to the normalization of disturbed circadian rhythms.
Two different approaches have been utilized to study the controlling mechanisms that underlie the generation and entrainment of circadian rhythms in mammals. The use of specific drugs to alter the period and/or the phase of circadian rhythms has provided new insights into both the pathways by which environmental information reaches the mammalian circadian pacemaker in the suprachiasmatic nuclei (SCN) and the cellular and neurochemical events within the SCN itself which are involved in circadian rhythmicity. A second approach, which seeks to exploit genetic differences in the properties of the circadian system, holds the promise of eventually defining the cellular and molecular events that are part of the clock itself, the events that underlie the entrainment of the circadian clock by environmental factors, and the expression of overt rhythms driven by the clock. It is anticipated that the pharmacological and genetic approaches to the study of circadian rhythms will complement each other as the underlying physiological mechanisms of the circadian clock system become defined.
While aging has been associated with changes in the period and amplitude of circadian rhythms, little is known about how aging influences the response of the circadian clock to environmental stimuli. In this paper, we report on recent studies designed to determine the effects of advanced age on the response of the circadian clock to both photic and nonphotic stimuli in old hamsters (e.g., over 16 mo of age). Among the most pronounced age-related changes in the circadian rhythm of locomotor activity are: (a) alterations in the phase-angle of entrainment to the light-dark cycle; (b) an increase in the magnitude of phase shifts induced by pulses of light presented at specific circadian times; and (c) a loss of responsiveness to the phase shifting or entraining effects of stimuli which induce an acute increase of activity. Depletion of brain monoamine levels in young animals can induce changes in the responsiveness of the circadian clock to environmental stimuli which are similar to those which occur spontaneously in old animals, suggesting that aging alters monoaminergic inputs to the clock. Some of the age-related changes in the response of the clock to an activity-inducing stimulus can be reversed by implanting old animals with fetal SCN tissue. Determining the physiological basis for age related changes in the responsiveness of the clock to both internal and external stimuli, and the mechanisms by which normal circadian function can be restored, should lead to new insight into the functioning of the circadian clock and may lead to new approaches for normalizing disturbed circadian rhythms.
Photostimulated male Djungarian hamsters following placement in a long-day photoperiod exhibit a characteristic rise in serum FSH levels that occurs in the absence of a simultaneous rise in LH levels. It is not known whether this singular FSH secretion is dependent upon a differential responsiveness of the gonadotrophs to the pattern of pulsatile GnRH release or is instead driven by a GnRH-independent mechanism. We have assessed the GnRH dependence of this singular FSH secretion by testing the ability of a potent GnRH antagonist (GnRHa: WY-45760) to block FSH and testicular responses to photostimulation. Photoinhibited hamsters were transferred from a short-day (6L:18D) to a long-day photoperiod (16L:8D). Hamsters received two daily injections of a GnRH antagonist or vehicle (VEH). After 0 (short day), 3, 5, 10, 30, or 40 days the hamsters were killed; plasma was assayed for FSH, LH, and testosterone (T), and testes weights were recorded. Testes were sectioned and analyzed for tubular development. In VEH-treated animals, testicular weights increased after photostimulation, reaching mean values of 514 mg by 30 days. Treatment with GnRHa resulted in a significant (p < 0.01) attenuation of testicular growth after 30 days of photostimulation (mean testes weight = 110.1 mg). In VEH-treated hamsters there was a rapid increase in FSH levels after photostimulation that became significant by 5 days and peaked at 10 days. In the GnRHa-treated group, however, these FSH increments were completely blocked at 5 days and significantly reduced at 10 days compared to the values in the corresponding VEH-treated groups.(ABSTRACT TRUNCATED AT 250 WORDS)
Serotonergic mechanisms are believed to play a considerable role in mediating the effects of photic and nonphotic stimuli on circadian rhythmicity. Because aging is associated with significant changes in the responsiveness of the rodent circadian system to major synchronizing agents in the environment, this study examined the phase-shifting effects of the 5-HT1A receptor agonist, 8-hydroxy-2-(di-n-propylamino)tetralin [8-OH-DPAT; 2.0, 5.0, and 8.0 mg/kg ip at circadian time 8 (CT 8)], on the 24-h activity rhythm of young (3-4 mo old) and old (18-19 mo old) golden hamsters. Aging was associated with a dramatic attenuation of the phase-shifting effects of 8-OH-DPAT in this species. The results suggest the existence of age-related deficits in the serotonergic control of mammalian circadian rhythmicity, which could interfere with the temporal adaptation of the senescent organism to its environment.
Noradrenergic locus ceruleus neurons are most active during waking and least active during rapid eye movement (REM) sleep. We expected REM sleep deprivation (REMSD) to increase norepinephrine utilization and activate the tyrosine hydroxylase (TH) gene critical for norepinephrine production. Male Wistar rats were deprived of REM sleep with the platform method. Rats were decapitated after 8, 24, or 72 h on small (REMSD) or large (control) platforms or after 8 or 24 h of rebound sleep after 72 h of the platform treatment. During the first 24 h, norepinephrine concentration, measured by high-performance liquid chromatography/electrochemical detection, was lower in the neocortex, hippocampus, and posterior hypothalamus in REMSD rats than in large-platform controls. After 72 h of REMSD, TH mRNA, measured by in situ hybridization, was increased in the locus ceruleus and norepinephrine concentrations were increased. Polygraphy showed that small-platform treatment caused effective and selective REMSD. Serum corticosterone measurement by radioimmunoassay indicated that the differences found in norepinephrine and TH mRNA were not due to differences in stress between the treatments. The novel finding of sleep deprivation-specific increase in TH gene expression indicates an important mechanism of adjusting to sleep deprivation.
A variety of stimuli, which are associated with acute increases in locomotor activity, induce similar phase-dependent shifts in the circadian rhythm of locomotor activity in golden hamsters. Treatment with the benzodiazepine, triazolam (Tz), or transfer of an animal normally housed without a running wheel to a new cage with 1 h of access to a wheel are both examples of such stimuli. Phase shifts normally induced by injections of Tz can be blocked by lesions of the intergeniculate leaflet (IGL) of the thalamus. Experiments were conducted to determine whether phase shifts induced by transfer to a new cage with a running wheel also require an intact IGL. Animals normally housed without running wheels were transferred to new cages with access to wheels for 1 h a few hours before the expected onset of activity. They then received either lesions of the IGL or sham lesions and, after recovery, they were again transferred to new cages with running wheels for 1 h. Lesions of the IGL blocked phase shifts normally induced by wheel access while sham lesions had no effect. The amount of wheel-running and total locomotor activity which occurred during access to the running wheel were significantly reduced by IGL lesions. These results indicate that the phase-shifting effect of a novel running wheel depends upon an intact IGL.
In a search for genes that regulate circadian rhythms in mammals, the progeny of mice treated with N-ethyl-N-nitrosourea (ENU) were screened for circadian clock mutations. A semidominant mutation, Clock, that lengthens circadian period and abolishes persistence of rhythmicity was identified. Clock segregated as a single gene that mapped to the midportion of mouse chromosome 5, a region syntenic to human chromosome 4. The power of ENU mutagenesis combined with the ability to clone murine genes by map position provides a generally applicable approach to study complex behavior in mammals.
In the present study, 18-25-month-old hamsters free-running in constant dim light were injected, both before and after receiving fetal grafts containing either cerebellar tissue or the suprachiasmatic nuclei (SCN), with a dose of triazolam given at a time known to reliably phase shift the rhythm of locomotor activity in young hamsters. SCN-grafted animals, but not control animals implanted with fetal cerebellar tissue, showed a significantly greater response to the phase shifting effects of triazolam, demonstrating that at least some age-related changes in the circadian system can be reversed by neuronal transplantation. These results raise the possibility that neuropharmacological interventions that can simulate the effects of fetal SCN grafts might be useful in the treatment of age-related disorders in circadian function.
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 monamine 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 +/- 17.7 min, P < 0.01). Our results suggest that monoaminergic systems are essential for the phase-shifting effect of Tz upon the circadian system in hamsters.
Changes in behavior which occur on a daily or circadian basis represent one of the most ubiquitous strategies by which most living organisms have adapted to their environment. Underlying the daily changes in behavior are a multitude of endocrine and metabolic rhythms which provide adaptively significant temporal organization within the organism. In mammals there appears to be a central circadian clock in the SCN which is responsible for generating and coordinating the entire 24-hour temporal organization of the animal. The circadian clock regulates the timing, duration, and characteristics of sleep, and together the circadian clock and sleep interact to control the timing of endocrine secretions. While the impact of disturbed endocrine circadian rhythms for the survival of the species has received very little attention, the almost universal presence of circadian rhythmicity within the endocrine system argues in support of the hypothesis that a disruption of the normal circadian organization within the endocrine system can have serious consequences for the health and well-being of the organism. It is particularly noteworthy that in advanced age, various alterations in circadian endocrine rhythms have been observed and these alterations may impair the ability of the animal to adapt normally to the environment. Relatively speaking, the study of circadian rhythms is a new field of biology, and as a result, much remains to be discovered about the physiological mechanisms that underlie rhythmicity, as well as the functional significance of 24-hour temporal organization for the survival of the species.
Reproductive activity in the Djungarian hamster is controlled by seasonal variations in day length. Exposure to long days stimulates testis development, while exposure to short days induces testis regression. We recently found that testis regression after gonadotropin deprivation in rats is associated with increases in apoptosis. Here we sought to determine whether or not apoptosis is associated with the testis regression and/or recrudescence that occurs naturally in seasonally breeding mammals. Newborn male hamsters were maintained on long days (16L:8D) until 3 wk of age before being transferred to short days (8L:16D). Following decreases in serum FSH within 3 days of exposure to short days, testis weight decreased by 52% at Day 10, reaching a 70% decrease after 21 days. Analysis of testis cell DNA fragmentation showed a 4.9-fold increase of low-molecular-weight DNA as early as 5 days after transfer to short days; this was followed by a time-dependent decrease. The observed increases in testis cell apoptosis were correlated with decreases in serum testosterone, but decreases in Leydig cell LH receptor content were delayed. In a second study, 6-wk-old hamsters with regressed testes due to a 3-wk exposure to short days were transferred back to long days. After increases in serum FSH within 3 days of photostimulation, a 2-fold elevation in testis weight was found at Day 5. The increase in testis weight was associated with a 65% decrease of testis apoptosis within 5 days of photostimulation. Also, increases in serum testosterone and LH receptor content were observed after 5 and 10 days of exposure to long days, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
To determine the magnitude and direction of phase shifts of human circadian rhythms occurring within 1 day after a single exposure to bright light, plasma thyrotropin, melatonin, and cortisol levels and body temperature were monitored for 38 h in 17 men who were each studied two times, once during continuous dim light conditions and once with light exposure. After a period of entrainment to a fixed sleep-wake cycle, a 3-h light pulse (5,000 lux) was presented under constant routine conditions, and the resultant phase shifts were measured, also under constant routine conditions, on the 1st day after pulse presentation. The phase shifts in response to light occurred within 24 h and were in the delaying direction for most of the nocturnal period, with the crossover to phase advances occurring approximately 1 h after the temperature minimum. Phase shifts averaged 1 h, with delays being larger than advances, and were achieved without significant changes in rhythm amplitude. The immediate response of the human circadian clock to a single 3-h light pulse is thus characteristic of "type 1" resetting.
To determine whether a single episode of physical activity is capable of inducing rapid phase shifts in human circadian rhythms, 17 subjects were studied two times under constant routine conditions, once in the absence of stimulus and once with a 3-h nighttime pulse of exercise interrupting the constant routine conditions. The profiles of plasma cortisol, thyrotropin (TSH), and melatonin and of body temperature were monitored continuously to derive estimations of circadian phase position. The phase shifts were measured on the 1st day after exercise exposure. The timing of the exercise period ranged from -5 h to +4 h around the time of the minimum body temperature rhythm. Nighttime exercise was associated with 1- to 2-h phase delays of both the melatonin and TSH rhythms, with the size of the delays tending to be smaller when the exercise was presented in the latter part of the nighttime period and in the early morning. These data demonstrate that nonphotic stimuli may exert phase-shifting effects on the human circadian pacemaker.
Melatonin, a pineal hormone, is known to play an important role in mediating changes in the reproductive system which occur in response to seasonal changes in the length of the day. An in vitro pituitary perifusion system has been used to examine both direct and indirect effects of melatonin on pituitary gonadotropin basal secretion and responsiveness to GnRH stimulation. Anterior pituitaries from adult male golden hamsters were perifused with an APS 10 perifusion system for 6 h. Both basal secretion and secretion in response to hourly GnRH stimulation (1 min, 35 ng/ml pulses) were evaluated for LH and FSH. In order to expose pituitaries to shorter and longer durations of melatonin, tissue was perifused with melatonin-containing medium following dissection in the daytime (when endogenous melatonin levels are low) and following dissection in the nighttime (when endogenous melatonin levels are high). Exposure of pituitary tissue from golden hamsters to melatonin during perifusion caused a decrease in basal secretion of LH but did not affect GnRH stimulated LH or FSH release regardless of the time of dissection. There was, however, an effect of some diurnal factor to lower FSH responsiveness to GnRH stimulation from tissue dissected in the nighttime. It is suggested that melatonin may be responsible for this diurnal difference. Therefore, melatonin can act directly at the anterior pituitary to lower basal LH secretion, and melatonin or some other diurnal factor can act in vivo to lower pituitary FSH secretion in response to GnRH stimulation during the night.