Multioscillator Circadian System Research Articles

Mechanical loading and hyperosmolarity as a daily resetting cue for skeletal circadian clocks

Daily rhythms in mammalian behaviour and physiology are generated by a multi-oscillator circadian system entrained through environmental cues (e.g. light and feeding). The presence of tissue niche-dependent physiological time cues has been proposed, allowing tissues the ability of circadian phase adjustment based on local signals. However, to date, such stimuli have remained elusive. Here we show that daily patterns of mechanical loading and associated osmotic challenge within physiological ranges reset circadian clock phase and amplitude in cartilage and intervertebral disc tissues in vivo and in tissue explant cultures. Hyperosmolarity (but not hypo-osmolarity) resets clocks in young and ageing skeletal tissues and induce genome-wide expression of rhythmic genes in cells. Mechanistically, RNAseq and biochemical analysis revealed the PLD2-mTORC2-AKT-GSK3β axis as a convergent pathway for both in vivo loading and hyperosmolarity-induced clock changes. These results reveal diurnal patterns of mechanical loading and consequent daily oscillations in osmolarity as a bona fide tissue niche-specific time cue to maintain skeletal circadian rhythms in sync.

Circadian Timing of the Female Reproductive System

Precise chronological transition is critical for a normal female reproductive system. Many epidemiologic studies have demonstrated rhythmicity in parturition during the resting phase, and even seasonal breeding. Circadian rhythms are controlled by a multi-oscillatory circadian system. These rhythms are determined by both genetic and environmental factors. The female reproductive axis is also highly rhythmic, and the hypothalamic-pituitary-gonadal axis and the hypothalamic-pituitary-adrenal axis are also functional multi-oscillatory circadian systems. Bidirectional communication between the central and peripheral tissue clocks is essential for maintaining the rhythm from ovulation to parturition. This review discusses the circadian timing of the female reproductive system, specifically its underlying metabolic and molecular clock functions. This review particularly focused on the following areas: multi-oscillatory system in the female reproductive system; the ovarian cycle and the timing of ovulation; timing of mating and seasonal reproduction; circadian rhythms in pregnancy; circadian timing of labor onset and parturition.

Circadian rhythm in mRNA expression of the glutathione synthesis gene Gclc is controlled by peripheral glial clocks in Drosophila melanogaster.

Circadian coordination of metabolism, physiology, and behaviour is found in all living kingdoms. Clock genes are transcriptional regulators, and their rhythmic activities generate daily rhythms in clock-controlled genes which result in cellular and organismal rhythms. Insects provide numerous examples of rhythms in behaviour and reproduction, but less is known about control of metabolic processes by circadian clocks in insects. Recent data suggest that several pathways involved in protecting cells from oxidative stress may be modulated by the circadian system, including genes involved in glutathione (GSH) biosynthesis. Specifically, rhythmic expression of the gene encoding the catalytic subunit (Gclc) of the rate-limiting GSH biosynthetic enzyme was detected in Drosophila melanogaster heads. The aim of this study was to determine which clocks in the fly multi-oscillatory circadian system are responsible for Gclc rhythms. Genetic disruption of tissue-specific clocks in D. melanogaster revealed that transcriptional rhythms in Gclc mRNA levels occur independently of the central pacemaker neurons, because these rhythms persisted in heads of behaviourally arrhythmic flies with a disabled central clock but intact peripheral clocks. Disrupting the clock specifically in glial cells abolished rhythmic expression of Gclc, suggesting that glia play an important role in Gclc transcriptional regulation, which may contribute to maintaining homeostasis in the fly nervous system.

The role of kisspeptin and RFRP in the circadian control of female reproduction

In female mammals, reproduction shows ovarian and daily rhythms ensuring that the timing of the greatest fertility coincides with maximal activity and arousal. The ovarian cycle, which lasts from a few days to a few weeks, depends on the rhythm of follicle maturation and ovarian hormone production, whereas the daily cycle depends on a network of circadian clocks of which the main one is located in the suprachiasmatic nuclei (SCN). In the last ten years, major progress has been made in the understanding of the neuronal mechanisms governing mammalian reproduction with the finding that two hypothalamic Arg-Phe-amide peptides, kisspeptin (Kp) and RFRP, regulate GnRH neurons. In this review we discuss the pivotal role of Kp and RFRP neurons at the interface between the SCN clock signal and GnRH neurons to properly time gonadotropin-induced ovulation. We also report recent findings indicating that these neurons may be part of the multi-oscillatory circadian system that times female fertility. Finally, we will discuss recent investigations indicating a role, and putative therapeutic use, of these neuropeptides in human reproduction.

A Multi-Oscillatory Circadian System Times Female Reproduction

Rhythms in female reproduction are critical to insure that timing of ovulation coincides with oocyte maturation and optimal sexual arousal. This fine tuning of female reproduction involves both the estradiol feedback as an indicator of oocyte maturation, and the master circadian clock of the suprachiasmatic nuclei (SCN) as an indicator of the time of the day. Herein, we are providing an overview of the state of knowledge regarding the differential inhibitory and stimulatory effects of estradiol at different stages of the reproductive axis, and the mechanisms through which the two main neurotransmitters of the SCN, arginine vasopressin, and vasoactive intestinal peptide, convey daily time cues to the reproductive axis. In addition, we will report the most recent findings on the putative functions of peripheral clocks located throughout the reproductive axis [kisspeptin (Kp) neurons, gonadotropin-releasing hormone neurons, gonadotropic cells, the ovary, and the uterus]. This review will point to the critical position of the Kp neurons of the anteroventral periventricular nucleus, which integrate both the stimulatory estradiol signal, and the daily arginine vasopressinergic signal, while displaying a circadian clock. Finally, given the critical role of the light/dark cycle in the synchronization of female reproduction, we will discuss the impact of circadian disruptions observed during shift-work conditions on female reproductive performance and fertility in both animal model and humans.

Putative pacemakers in the eyestalk and brain of the crayfish Procambarus clarkii show circadian oscillations in levels of mRNA for crustacean hyperglycemic hormone.

Crustacean hyperglycemic hormone (CHH) synthesizing cells in the optic lobe, one of the pacemakers of the circadian system, have been shown to be present in crayfish. However, the presence of CHH in the central brain, another putative pacemaker of the multi-oscillatory circadian system, of this decapod and its circadian transcription in the optic lobe and brain have yet to be explored. Therefore, using qualitative and quantitative PCR, we isolated and cloned a CHH mRNA fragment from two putative pacemakers of the multi-oscillatory circadian system of Procambarus clarkii, the optic lobe and the central brain. This CHH transcript synchronized to daily light-dark cycles and oscillated under dark, constant conditions demonstrating statistically significant daily and circadian rhythms in both structures. Furthermore, to investigate the presence of the peptide in the central brain of this decapod, we used immunohistochemical methods. Confocal microscopy revealed the presence of CHH-IR in fibers and cells of the protocerebral and tritocerebal clusters and neuropiles, particularly in some neurons located in clusters 6, 14, 15 and 17. The presence of CHH positive neurons in structures of P. clarkii where clock proteins have been reported suggests a relationship between the circadian clockwork and CHH. This work provides new insights into the circadian regulation of CHH, a pleiotropic hormone that regulates many physiological processes such as glucose metabolism and osmoregulatory responses to stress.

DISSOCIATION OF THE CIRCADIAN SYSTEM OF OCTODON DEGUS BY T28 AND T21 LIGHT-DARK CYCLES

Octodon degus is a primarily diurnal rodent that presents great variation in its circadian chronotypes due to the interaction between two phase angles of entrainment, diurnal and nocturnal, and the graded masking effects of environmental light and temperature. The aim of this study was to test whether the circadian system of this diurnal rodent can be internally dissociated by imposing cycles shorter and longer than 24 h, and to determine the influence of degus chronotypes and wheel-running availability on such dissociation. To this end, wheel-running activity and body temperature rhythms were studied in degus subjected to symmetrical light-dark (LD) cycles of T28h and T21h. The results show that both T-cycles dissociate the degus circadian system in two different components: one light-dependent component (LDC) that is influenced by the presence of light, and a second non–light-dependent component (NLDC) that free-runs with a period different from the external lighting cycle. The LDC was more evident in the nocturnal than diurnal chronotype, and also when wheel running was available. Our results show that, in addition to rats and mice, degus must be added to the list of species that show an internal dissociation in their circadian rhythms when exposed to forced desynchronization protocols. The existence of a multioscillatory circadian system having two groups of oscillators with low coupling strength may explain the flexibility of degus chronotypes. (Author correspondence: angerol@um.es)

Sleep and Circadian Rhythms in Humans

During the past 50 years, converging evidence reveals that the fundamental properties of the human circadian system are shared in common with those of other organisms. Concurrent data from multiple physiological rhythms in humans revealed that under some conditions, rhythms oscillated at different periods within the same individuals and led to the conclusion 30 years ago that the human circadian system was composed of multiple oscillators organized hierarchically; this inference has recently been confirmed using molecular techniques in species ranging from unicellular marine organisms to mammals. Although humans were once thought to be insensitive to the resetting effects of light, light is now recognized as the principal circadian synchronizer in humans, capable of eliciting weak (Type 1) or strong (Type 0) resetting, depending on stimulus strength and timing. Realization that circadian photoreception could be maintained in the absence of sight was first recognized in blind humans, as was the property of adaptation of the sensitivity of circadian photoreception to prior light history. In sighted humans, the intrinsic circadian period is very tightly distributed around approximately 24.2 hours and exhibits aftereffects of prior entrainment. Phase angle of entrainment is dependent on circadian period, at least in young adults. Circadian pacemakers in humans drive daily variations in many physiologic and behavioral variables, including circadian rhythms in alertness and sleep propensity. Under entrained conditions, these rhythms interact with homeostatic regulation of the sleep/wake cycle to determine the ability to sustain vigilance during the day and to sleep at night. Quantitative understanding of the fundamental properties of the multioscillator circadian system in humans and their interaction with sleep/wake homeostasis has many applications to health and disease, including the development of treatments for circadian rhythm and sleep disorders.

Circadian rhythm of ERG in Iguana iguana: role of the pineal.

In green iguanas, the pineal controls the circadian rhythm of body temperature but not the rhythm of locomotor activity. As part of a program to investigate the characteristics of this multioscillator circadian system, the authors studied the circadian rhythms of the electroretinographic response (ERG) and asked whether the pineal gland is necessary for the expression of this rhythm. ERGs from a total of 24 anesthetized juvenile iguanas were recorded under four different conditions: (a) complete darkness (DD), (b) dim light-dark cycles (dLD), (c) constant dim light (dLL), and (d) pinealectomized in DD. Results demonstrate that the b-wave component of the ERG shows a very clear circadian rhythm in DD and that this rhythm persists in dLL and entrains to dLD cycles. The ERG response is maximally sensitive during the subjective day. Pinealectomy does not abolish the circadian rhythm in ERG, demonstrating that the oscillator responsible for the ERG rhythm is located elsewhere.

Light-induced uncoupling of multioscillatory circadian system in a diurnal rodent, Asian chipmunk.

Responses of the circadian locomotor rhythm to a single light pulse were examined in a diurnal rodent, Asian chipmunk, by exposing it to a 1-h light pulse of 2,000 lx under constant conditions. A light pulse given at the beginning and end of the subjective night produced a phase delay and advance shifts, respectively. When pulsed around the midpoint of the subjective night, the circadian rhythm was shifted as much as 12 h in most animals or became arrhythmic in some. In the latter case, an additional light pulse restored the circadian rhythm. Some animals were unresponsive to light. The phase response curve is categorized as type 0. A large phase-shift was sometimes followed by splitting of an activity band into two components. These results are best explained by an assumption that the chipmunk circadian system is composed of two mutually coupled major oscillators, each of which is constituted by multiple oscillators. Our results suggest that light affects the oscillatory coupling not only of the major oscillators but also of constitutional oscillators.

S4-3 Cellular basis of multioscillatory circadian system in the brain

In this paper, a simplified nonlinear method is proposed to enhance the transient stability of multimachine power system by using a Static Synchronous Series Compensator (SSSC). The rate of dissipation of transient energy is used to determine the additional damping provided by a SSSC. The proposed algorithm is based on the direct Lyapunov method. The simplicity of the proposed scheme and its robustness with respect to large disturbances constitute the main positive features. Simulation results in the case of 3-machines power system show the effectiveness of the proposed method under large disturbances (3-phase and single phase short-circuits).

S4-3 Cellular basis of multioscillatory circadian system in the brain

S4-3 Cellular basis of multioscillatory circadian system in the brain

Anticipatory activity rhythms under daily schedules of water access in the rat.

Rats anticipate a fixed daily feeding time by entrainment of a component of their multioscillatory circadian system. The range of stimuli capable of entraining this component is little studied. Previous studies suggest that restricted water access is not an effective entrainment stimulus, as measured by general locomotion. The present study re-examined the issue, using two other measures of activity: wheel running and activity at a food-water delivery bin. Rats restricted to 1 hr of water each day in the middle of the light and to food in the 12-hr dark period showed no anticipation of this event in the wheel-running measure, but some rats did show anticipation in the delivery bin activity measure. Rats (bin activity measure only) restricted to 1 hr of water and 1 hr of food separated by intervals of 7, 10, or 12 hr, in either the light or the dark, showed consistent anticipation of food access time but little or no anticipation of water access time. Water access time also did not sustain food anticipatory rhythms in animals whose food-water schedules were reversed. However, deprivation of water or of both food and water for 72 or 90 hr was usually associated with specific increases in bin activity at both the usual feeding and drinking times. Water access, like food, appears to provide cues capable of entraining an anticipatory circadian mechanism. Differences in the type and amount of anticipatory activity preceding these events may reflect differences in the strengths of the two entrainment cues and/or in the activity levels or specific behavioral strategies promoted by hunger and thirst.

Retinally perceived light can entrain the pineal melatonin rhythm in Japanese quail

The avian pineal organ contains a circadian oscillator that can drive a daily rhythm of melatonin synthesis. In some avian species the pineal organ may act, via the cyclic release of melatonin, as a pacamaker within a multioscillator circadian system. The routes by which light entrains the pineal melatonin rhythm were investigated in the Japanese quail. A ‘patching’ protocol was used to expose directly either the eyes or the pineal to a light-dark cycle while the rest of the bird was exposed to constant light. The results show that the pineal melatonin rhythm can be entrained (1) by light perceived directly or (2) by light perceived by the eyes. Furthermore, the pathway by which light entrains the pineal melatonin rhythm includes the optic nerves because transection of the optic nerve eliminates the ability of ocularly perceived light to entrain the pineal melatonin rhythm.

Pineal melatonin rhythms in the lizard Anolis carolinensis: I. Response to light and temperature cycles.

Both light and temperature can influence the pineal's synthesis of the indoleamine melatonin. An investigation of the effects of light and temperature cycles on the pineal melatonin rhythm (PMR) showed the following: (1) Both daily light cycles and daily temperature cycles could entrain the PMR; melatonin levels peaked during the dark phase of a light-dark cycle or the cool phase of a temperature cycle. (2) The PMR could be entrained by a temperature cycle as low as 2 degrees C in amplitude in lizards held in constant light or constant darkness. (3) The length of the photoperiod or thermoperiod affected the phase, amplitude, or duration of the PMR. (4) When presented together, the effects of light and temperature cycles on the PMR depended on the phase relationship between the light and temperature cycles, as well as on the strength of the entraining stimuli, such as the amplitude of the temperature cycle. (5) Exposure to a constant cold temperature (10 degrees C) eliminated the PMR, yet a rhythm could still be expressed under a 24-hr temperature cycle (32 degrees C/10 degrees C), and the rhythm peaked during the 10 degrees C phase of the cycle. (6) A 6-hr dark pulse presented during the day did not elicit a premature rise in melatonin levels. These studies show how environmental stimuli can control the pineal rhythm of melatonin synthesis and secretion. Previous studies have supported a model in which the lizard's pineal acts as a circadian pacemaker within a multioscillator circadian system, and have implicated melatonin as a hormone by which the pineal may communicate with the rest of the system. The lizard pineal, therefore, may act as a photo- and thermoendocrine transducer translating light and temperature information into an internal cue in the form of the PMR. The PMR, in turn, may control the phase and period of circadian clocks located elsewhere, insuring that the right internal events occur at the right time of day.

Interaction between light- and feeding-entrainable circadian rhythms in the rat

The interaction between light-entrainable rhythms (LER) and feeding-entrainable rhythms (FER) of wheel running was studied by maintaining 16 rats in constant darkness and on ad lib feeding for 14 days, followed by a restricted feeding schedule (2 hr food/day) for 42 days, and 25 days on ad lib feeding. In 4 rats the LER assumed the same period as food access during restricted feeding. The pre-restriction periods of the LER of these rats differed from that of the feeding schedule by 5 min or less. However, since food access did not determine the phase of the LER, food does not appear to be a true Zeitgeber for this rhythm. No synchronization was observed when the two periods differed by more than 10 min, although changes in period or phase of the LER during restricted feeding were observed in all rats. Four hour phase advances or delays of the feeding schedule did not result in consistent effects on the phase of the LER. Two hour light pulses near the onset or end of the LER resulted in phase delays and phase advances of the LER, but had no effect on the FER. In the context of a multioscillator circadian system, these results suggest that light- and feeding-entrainable circadian oscillators are only weakly coupled.

Entrainment of the circadian activity rhythm of a lizard to melatonin injections

The circadian activity rhythms of lizards (Sceloporus occidentalis) can be entrained (synchronized) to a period of 24 hr by melatonin injections given every other day at the same time of day, but not by saline injections. The activity onsets of the entrained lizards exhibited two preferred phase-relationships (∼165° and ∼30°) with the time of melatonin injections with the 30° phase only rarely observed. These results suggest that endogenous rhythms of melatonin secretion (i.e., from the pineal organ) may be involved in synchronizing circadian oscillations within the lizard's multioscillator circadian system.

Circadian organization in Japanese quail.

Our recent studies have implicated both the eyes and pineal as major components of the circadian system of Japanese quail. We assessed the role of these organs by examining the effect of their removal on the circadian activity rhythm of quail exposed to either 24 hr light-dark (LD) cycles or to continuous darkness (DD). Removal of only the pineal had no effect on the activity rhythm of quail in either LD or DD. Blinding (by orbital enucleation) had a major effect under both LD and DD. One third of the blinded birds showed entrainment under LD although entrainment patterns were very variable, whereas two thirds of blinded birds were arrhythmic. All blinded plus pinealectomized birds were arrhythmic in LD as were all blinded and blinded plus pinealectomized birds in DD. Accordingly, effects of pinealectomy can be seen only when pinealectomy is combined with blinding. The fact that blinding disrupts circadian organization in both LD and DD indicates that the eyes must act as major components of the quail's circadian system. In view of the postulated role for melatonin, an indoleamine, in circadian systems, the eyes, pineal, and blood of quail were assayed for this compound. Robust daily rhythms in melatonin content were observed in all three tissues. The blood rhythm is due to secretion of melatonin into the vascular system by both the pineal and eyes. The ocular melatonin rhythm continued after sectioning of the optic nerve, was reentrainable to a shift in the phase of the LD cycle, and persisted for at least 2 days in DD. These data suggest that the eyes play a major role within the circadian system and support the hypothesis that circadian pacemakers may reside within the eyes of quail. The results are discussed in view of the findings of others in both quail and other avian species. A general model for circadian organization in birds is presented in which the eyes, the pineal, and the suprachiasmatic nuclei of the hypothalamus comprise major elements of a multioscillator circadian system.

A comparison of light and temperature entrainment: evidence for a multioscillator circadian system

ABSTRACT. Both photoperiodic and thermoperiodic cycles synchronize circadian calling activity of male crickets, Teleogryllus commodus (Walker). Because the phase relationships between these cycles and the entrained singing activity are clearly distinguishable, we studied the relative power of these factors in affecting the circadian clock. Upon resumption of constant conditions after exposure to a thermoperiodic cycle, singing activity sometimes splits into two daily bouts, each with a distinctive period. This observation, together with results derived from simultaneous fluctuation of both factors in and out of phase, suggests that the system integrating environmental input is composed of multiple oscillators.

Neurobiology of Circadian Rhythms in Mammals

The discovery in 1972 that the hypothalamic suprachiasmatic nucleus (SCN) is a major component of the multioscillatory circadian system in mammals provided biologists with a focal point for the study of the neural basis of circadian rhythms. Although the SCN's precise role in the generation of circadian rhythms is still not understood, recent studies of this nucleus have laid the groundwork for the eventual elucidation of the neural basis for: the generation of circadian signals, the interaction of circadian oscillators, the regulation of circadian oscillators by internal and external environmental factors, and the regulation of the temporal pattern of a variety of cellular, physiological, and behavioral events by circadian pacemaking units in mammals. (Accepted for publication 16 January 1983)