Main Clock Research Articles

A 3.5 GHz Spread-Spectrum Clock Generator With a Memoryless Newton-Raphson Modulation Profile

A frequency-locked loop (FLL) based spread-spectrum clock generator (SSCG) with a memoryless Newton-Raphson modulation profile is introduced in this paper. The SSCG uses an FLL as a main clock generator. It brings not only an area reduction to the SSCG but also the advantage of having multiple frequency deviations. A double binary-weighted DAC is proposed that modulates the frequency information of the frequency detector using a 1-1-1 MASH <formula formulatype="inline" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex Notation="TeX">$\Delta \Sigma$</tex></formula> modulator. The Newton-Raphson mathematical algorithm is applied to the proposed profile generator in order to generate the optimized nonlinear profile without needing any memory, resulting in a reduction in the area and the power consumption. It also makes it possible to have multiple modulation frequencies. The SSCG can support 14 frequency deviations of <formula formulatype="inline" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex Notation="TeX">$\pm 0.5\%$</tex></formula> to 3.5% in steps of 0.5% and three modulation frequencies of <formula formulatype="inline" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex Notation="TeX">$f _{\rm m}$</tex></formula> , <formula formulatype="inline" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex Notation="TeX">$2 f _{\rm m}$</tex></formula> and <formula formulatype="inline" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex Notation="TeX">$3 f _{\rm m}$</tex></formula> . It achieved an EMI reduction of 19.14 dB with a 0.5% down spreading and a 31 kHz modulation frequency, while employing a core area of 0.076 <formula formulatype="inline" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex Notation="TeX">${\hbox{mm}}^{2}$</tex> </formula> in a 0.13- <formula formulatype="inline" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"><tex Notation="TeX">$\mu{\hbox{m}}$</tex> </formula> CMOS process and consuming 23.72 mW at 3.5 GHz.

Tuning the Mammalian Circadian Clock: Robust Synergy of Two Loops

The circadian clock is accountable for the regulation of internal rhythms in most living organisms. It allows the anticipation of environmental changes during the day and a better adaptation of physiological processes. In mammals the main clock is located in the suprachiasmatic nucleus (SCN) and synchronizes secondary clocks throughout the body. Its molecular constituents form an intracellular network which dictates circadian time and regulates clock-controlled genes. These clock-controlled genes are involved in crucial biological processes including metabolism and cell cycle regulation. Its malfunction can lead to disruption of biological rhythms and cause severe damage to the organism. The detailed mechanisms that govern the circadian system are not yet completely understood. Mathematical models can be of great help to exploit the mechanism of the circadian circuitry. We built a mathematical model for the core clock system using available data on phases and amplitudes of clock components obtained from an extensive literature search. This model was used to answer complex questions for example: how does the degradation rate of Per affect the period of the system and what is the role of the ROR/Bmal/REV-ERB (RBR) loop? Our findings indicate that an increase in the RNA degradation rate of the clock gene Period (Per) can contribute to increase or decrease of the period - a consequence of a non-monotonic effect of Per transcript stability on the circadian period identified by our model. Furthermore, we provide theoretical evidence for a potential role of the RBR loop as an independent oscillator. We carried out overexpression experiments on members of the RBR loop which lead to loss of oscillations consistent with our predictions. These findings challenge the role of the RBR loop as a merely auxiliary loop and might change our view of the clock molecular circuitry and of the function of the nuclear receptors (REV-ERB and ROR) as a putative driving force of molecular oscillations.

2,2,2-Tribromoethanol Phase-Shifts the Circadian Rhythm of the Liver Clock in Per2::Luciferase Knockin Mice: Lack of Dependence on Anesthetic Activity

Comprehensive gene expression profiling in mice in response to the inhalation of sevoflurane has revealed that circadian clock gene expression is affected strongly in the liver, heart, lung, and kidney, in this order, but moderately in the spleen and slightly in the brain. Therefore, we examined whether the administration of general anesthetics at different times of the day induces phase shifts of the liver clock in Per2::Luciferase knockin mice. One to 4 days of intraperitoneal injection of 2,2,2-tribromoethanol (240 mg/kg, anesthetic time 60 min) or 2,2,2-trichloroethanol (240 mg/kg, 60 min), common anesthetics in veterinary surgery, caused phase delays when injected during the daytime and phase advances when injected during the nighttime. Inhalation administration of isoflurane for 30 or 60 min during the daytime did not induce a phase delay. Injection of propofol (300 mg/kg, 17 min) during the daytime induced an insignificant phase delay of the Per2 bioluminescence rhythm. Injection of 2,2,2-tribromoethanol did not induce a phase shift in the suprachiasmatic nucleus, the main oscillator, or in behavioral locomotor rhythms, suggesting that 2,2,2-tribromoethanol induced phase shifts of the liver clock independent of the main suprachiasmatic clock. The expression of clock genes, such as Bmal1 and Clock, in mouse liver was decreased strongly 1 and 4 h after a single injection of 2,2,2-tribromoethanol. These results demonstrate that 2,2,2-tribromoethanol or 2,2,2-trichloroethanol produce phase shifts of the peripheral clock, independent of anesthetic activity. These anesthetics may cause circadian rhythm disorders in peripheral organs when administered as general anesthetics several times during the day.

Interactions between metabolism and circadian clocks: reciprocal disturbances

Obesity is a medical condition of excess body fat, recognized as a global epidemic. Besides genetic factors, overconsumption of high-energy food and a sedentary lifestyle are major obesogenic causes. A newly identified determinant is altered circadian rhythmicity. To anticipate and adapt to daily changes in the environment, organisms have developed an endogenous circadian timing system, comprising a main circadian clock, located in the suprachiasmatic nucleus (SCN) of the hypothalamus, principally synchronized to the light-dark cycle. Secondary peripheral clocks are found in various tissues, such as the liver, pancreas, and adipose tissue. These clocks control the rhythmic patterns of myriad metabolic processes. We will review the evidence that metabolic dysfunction is associated with circadian disturbances at both central and peripheral levels and, conversely, that disruption of circadian clock functioning can lead to obesity. The roots of these reciprocal interactions will be illustrated by transcriptional crosstalk between metabolic and circadian systems. Chronotherapeutic approaches of dieting to maintain or restore a proper circadian alignment could be useful to limit the magnitude of metabolic risks.

Quartz Resonator Based, 0.12 μW, 32768 Hz Oscillator with ±100 ppm Frequency Accuracy

A 0.12 μW power dissipation quartz oscillator with 32,768 Hz frequency was designed and fabricated. Stability of the oscillator versus power supply and temperature variations was measured. The design is suitable for the role of the RTC (real-time clock) or main system clock in low-power, battery-powered and energy harvesting systems.

Alteration of Hypothalamic–Pituitary–Thyroid Axis Function in Non-Small-Cell Lung Cancer Patients

The aim of this study was to evaluate the hypothalamic-pituitary-thyroid (HPT) axis function in patients suffering from lung cancer. Thyrotropin-releasing hormone (TRH), thyroid-stimulating hormone (TSH), free thyroxine (FT4), interleukin (IL)-2, and melatonin serum levels were measured in blood samples collected every 4 hours for 24 hours from 11 healthy participants (H; ages 35-53 years) and 9 patients suffering from non-small-cell lung cancer (C; ages 43-63 years). Relationships between hormone levels overall and over time of day were evaluated within and among groups. A prominent circadian rhythm with peaks near midnight was present for TSH and melatonin serum levels in both H and C, indicating similar synchronization of the main body clock to the 24-hour environmental light-dark cycle. As regards 24-hour means in H and C, TSH was lower in C, whereas TRH, FT4, and IL-2 were higher in C, with no difference in melatonin levels. Simple linear regression, FT4 versus TRH, showed a positive correlation in H and a negative correlation in C, whereas FT4 versus TSH showed a negative correlation in both groups. For FT4 versus IL-2, a negative correlation was found in C but not for H, whereas TSH versus TRH showed no correlation for either group. Both groups were found to be similarly synchronized to the 24-hour sleep-wake schedule, but HPT axis function was altered in patients suffering from lung cancer. When compared with healthy controls, cancer patients showed modifications of hormone serum levels overall and a negative correlation between individual TRH and FT4 levels.

High speed column-parallel CDS/ADC circuit with nonlinearity compensation for CMOS image sensors

A high speed column-parallel CDS/ADC circuit with nonlinearity compensation is proposed in this paper. The correlated double sampling (CDS) and analog-to-digital converter (ADC) functions are integrated in a three-phase column-parallel circuit based on two floating gate inverters and switched-capacitor network. The conversion rate of traditional single-slope ADC is speeded up by dividing quantization to coarse step and fine step. A storage capacitor is used to store the result of coarse step and locate the section of ramp signal of fine step, which can reduce the clock step from 2 n to 2(n/2+1). The floating gate inverters are implemented to reduce the power consumption. Its induced nonlinear offset is cancelled by introducing a compensation module to the input of inverter, which can equalize the coupling path in three phases of the proposed circuit. This circuit is designed and simulated for CMOS image sensor with 640×480 pixel array using Chartered 0.18 μm process. Simulation results indicate that the resolution can reach 10-bit and the maximum frame rate can reach 200 frames/s with a main clock of 10 MHz. The power consumption of this circuit is less than 36.5 μW with a 3.3 V power supply. The proposed CDS/ADC circuit is suitable for high resolution and high speed image sensors.

Effects of PACAP on the oxidative stress-induced cell death in chicken pinealocytes is influenced by the phase of the circadian clock

Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide with highly potent neuro- and general cytoprotective actions. PACAP is also an important modulator of circadian rhythmic functions, including time-dependent effects in the pineal gland. It is not known whether PACAP influences the survival of pinealocytes. The present study had two aims. First, we tested whether the cytoprotective effects of PACAP are present also in the pineal cells. As the pineal gland is the main circadian master clock in birds, we also tested whether this effect depends on the time of day. Using flow cytometry, we detected a significant decrease of cell viability after hydrogen peroxide-induced oxidative stress in chicken pinealocytes. PACAP alone did not influence cell survival. Co-incubation with PACAP in the dark phase (9pm) was able to attenuate the toxic effect of H2O2. The survival-promoting effect could be counteracted by simultaneously applied PACAP antagonist, PACAP6-38. However, co-treatment with PACAP during the light phase (9am) did not result in significant differences in the percentage of living cells. In summary, our results show that PACAP has a protective effect against the oxidative stress-induced cell death in chicken pinealocytes, but this effect is dependent on the phase of the circadian biological clock.

Behavioural food anticipation in clock genes deficient mice: confirming old phenotypes, describing new phenotypes

Animals fed daily at the same time exhibit circadian food-anticipatory activity (FAA), which has been suggested to be driven by one or several food-entrainable oscillators (FEOs). FAA is altered in mice lacking some circadian genes essential for timekeeping in the main suprachiasmatic clock (SCN). Here, we confirmed that single mutations of clock genes Per1(-/-) and Per2(Brdm1) alter FAA expression in constant darkness (DD) or under a light-dark cycle (LD). Furthermore, we found that Per1(-/-);Per2(Brdm1) and Per2(Brdm1);Cry1(-/-) double mutant animals did not display a stable and significant FAA either in DD or LD. Interestingly, rescued behavioural rhythms in Per2(Brdm1);Cry2(-/-) mice in DD were totally entrained to feeding time and re-synchronized after phase-shifts of mealtime, indicating a higher SCN sensitivity to feeding cues. However, under an LD cycle and restricted feeding at midday, FAA in double Per2(Brdm1);Cry2(-/-) mutant mice was absent. These results indicate that shutting down one or two clock genes results in altered circadian meal anticipation. Moreover, we show that in a genetically rescued SCN clock (Per2(Brdm1);Cry2(-/-)), food is a powerful zeitgeber to entrain behavioural rhythms, leading the SCN to be more sensitive to feeding cues than in wild-type littermates.

The neuronal network of the endogenous clock

Abstract Endogenous clocks control the rhythm of many biological processes. Malfunction of endogenous clocks in humans can lead to various diseases as sleep disorders, depres­sions, the metabolic syndrome and cancer. All animals have a main clock in the brain. This clock comprises a network of clock neurons that communicate with each other. In each clock neuron, conserved clock genes and pro­teins interact in to generate a molecular os­cillation. The molecular basis of this rhythm generation as well as the anatomy of the neuronal clock network is best investigated in the fruit fly Drosophila melanogaster. In the little fly, clock genes can be shut down in specific clock neurons. Furthermore, specific clock neurons can be electrically silenced and the rhythmic behaviour of such manipulated flies can be studied. A flurry of recent studies has begun to identify the role of specific clock neurons in the clock network, and these find­ings are helping to understand the basic neu­ronal mechanisms of endogenous clocks.

Lose Sleep, Gain Weight: Another Piece of the Obesity Puzzle

It’s 11 p.m., and you sit in front of a glowing computer screen, writing e-mails and eating a sandwich. You’ll work until after midnight, when you’ll fall asleep in front of the light and blare of a TV before rising again at 6 a.m. What’s wrong with this picture? Because of modern conveniences and pressures, many of us keep our bodies exposed to light, food, and activity at times when our organs and cells expect dark, quiet, and sleep. In epidemiologic studies, shorter sleep has been correlated with incidence of obesity, hypertension, and other metabolic disorders. Experimental sleep studies find a similar connection. Increasingly, studies of the possible mechanisms behind these associations suggest that lack of sleep is part of a bigger problem with the 24/7 lifestyle many people today lead. Increasingly, scientists are finding that many physiologic activities related to metabolism don’t happen continuously but oscillate on a regular schedule. Studies in mice as well as humans suggest that when our internal clock is disrupted, it may throw off many bodily functions, especially metabolism. Many environmental factors have been shown to contribute to circadian disruption. Noise in busy hospitals, street noise, and airport noise have all been reported to disrupt sleep or reduce its quality. Research in animals and humans shows that exposure to light during early biological night resets the main circadian clock by producing a phase delay (the biological urge to go to sleep and wake up later than usual), and exposure during late biological night results in a phase advance (going to sleep and waking up earlier than usual). We live in a world where air passengers can see the glow of major cities 200 miles away. So the fact that human circadian systems appear sensitive even to low-level artificial light exposure raises significant concerns for the health effects of our electrified modern society. For instance, exposure to a few hours of ordinary room light of about 100 lux brightness (which most people get every night before they go to bed) can significantly reset the human circadian pacemaker, Jamie Zeitzer and colleagues reported in the September 2005 American Journal of Physiology—Regulatory, Integrative and Comparative Physiology. However, it’s not certain what the effects of very brief light exposures may be. The duration of light exposure needed to cause shifts hasn’t been well studied, according to a review by Charles Czeisler and Joshua Gooley in volume 27 (2007) of Cold Spring Harbor Symposia on Quantitative Biology. [For more information about the health effects of too much artificial light, see “Missing the Dark: Health Effects of Light Pollution,” EHP 117:A20–A27 (2009).] Epidemiologic studies suggest that lack of sleep or sleeping on an altered schedule is an independent risk factor for gaining weight. But it’s still not certain whether short sleep actually causes obesity and its associated health effects. For instance, some scientists have suggested that the association between obesity and lack of sleep may be due to the fact that people who are obese may be more likely to have a sleep disorder such as sleep apnea, or that the reported lack of sleep is a symptom of psychosocial stress. But a body of studies have shown a connection between short sleep and obesity, other health effects associated with obesity, and increased appetite or food intake.

Neurogenetics of food anticipation

Circadian clocks enable the organisms to anticipate predictable cycling events in the environment. The mechanisms of the main circadian clock, localized in the suprachiasmatic nuclei of the hypothalamus, involve intracellular autoregulatory transcriptional loops of specific genes, called clock genes. In the suprachiasmatic clock, circadian oscillations of clock genes are primarily reset by light, thus allowing the organisms to be in phase with the light-dark cycle. Another circadian timing system is dedicated to preparing the organisms for the ongoing meal or food availability: the so-called food-entrainable system, characterized by food-anticipatory processes depending on a circadian clock whose location in the brain is not yet identified with certainty. Here we review the current knowledge on food anticipation in mice lacking clock genes or feeding-related genes. The food-entrainable clockwork in the brain is currently thought to be made of transcriptional loops partly divergent from those described in the light-entrainable suprachiasmatic nuclei. Possible confounding effects associated with behavioral screening of meal anticipation in mutant mice are also discussed.

Synchronization in a Multilevel CMOS Time-to-Digital Converter

Accurate time-to-digital conversion is typically based on determining the positions of the timing signals within the period of an accurate clock with digital delay-line interpolators. In order to save circuit area and to improve single-shot precision to the picosecond level, multilevel interpolators can be used. Timing signals are generally asynchronous with respect to the main clock, and thus, in order to obtain unambiguous and errorless results, careful attention should be given to the synchronization of the timing signals and various operating blocks and to the generation of the interpolation residue between the interpolators. This paper attempts to describe these problems in detail and suggests some solutions using a time-to-digital converter architecture based on two-level interpolation as a test vehicle, which demonstrates 6-ps rms single-shot precision in a measurement range of 1 ms.

New light on the serotonergic paradox in the rat circadian system

The main mammalian circadian clock, localized in the suprachiasmatic nuclei can be synchronized not only with light, but also with serotonergic activation. Serotonergic agonists and serotonin reuptake inhibitors (e.g., fluoxetine) have a non-photic influence (shifting effects during daytime and attenuation of photic resetting during nighttime) on hamsters' and mice' main clock. Surprisingly, in rats serotonergic modulation of the clock shows essentially photic-like features in vivo (shifting effects during nighttime). To delineate this apparent paradox, we analyzed the effects of fluoxetine and serotonin agonists on rats' clock. First, fluoxetine induced behavioral phase-advances associated with down-regulated expression of the clock genes Per1 and Rorbeta and up-regulated expression of Rev-erbalpha during daytime. Moreover, fluoxetine produced an attenuation of light-induced phase-advances in association with altered expression of Per1, Per2 and Rorbeta during nighttime. Second, we showed that 5-HT(1A) receptors -maybe with co-activation of 5-HT(7) receptors- were implicated in non-photic effects on the main clock. By contrast, 5-HT(3) and 5-HT(2C) receptors were involved in photic-like effects and, for 5-HT(2C) subtype only, in potentiation of photic resetting. Thus this study demonstrates that as for other nocturnal rodents, a global activation of the serotonergic system induces non-photic effects in the rats' clock during daytime and nighttime.

Pseudorandom signal sampler for relaxed design of multistandard radio receiver

This paper proposes a novel software defined radio (SDR) receiver design using non-uniform sampling (NUS) technique implemented by original design of a pseudorandom signal sampler (PSS) circuit for controlling data conversion to relax multistandard receiver circuit constraints. The proposed and designed NUS-based SDR receiver allows spectral alias suppression at integer multiples of sampling frequency offering the advantages of relaxing anti-aliasing filter (AAF), reducing the analog-to-digital converter (ADC) dynamic power consumption and the automatic gain control (AGC) range as well. The PSS circuit, generating pseudorandom clock signal, with enough time-quantization accuracy, was designed. The PSS is implemented in 65-nm digital CMOS technology and occupies 470(μm)2. It features up to 200MHz “mean clock” for 3.2GHz main clock while drawing 242μA for 1.2V supply. Mixed experimental/simulation tests, of designed NUS-based SDR receiver, revealed a confirmation of alias-free performances and the achievement of a 72dB (12-bit ADC) dynamic range after signal reconstruction.

Entrainment and coupling of the hamster suprachiasmatic clock by daily dark pulses

The circadian rhythm of locomotor activity of hamsters kept in constant light (LL) can split into two distinct components that, in steady state, lie 180 degrees apart. The splitting phenomenon is the result of antiphase circadian oscillations between left and right sides of the suprachiasmatic nuclei (SCN), the master circadian clock in mammals. In unsplit hamsters housed in LL, a single dark pulse produces a phase-shift of the wheel-running activity rhythm, accompanied by a transient down-regulation of clock gene expression in the SCN. In the present study, we evaluated the effects of daily 1-hr dark pulses on wheel-running activity rhythm and on the expression of clock and nonclock proteins in the SCN of Syrian hamsters exposed to LL conditions. The results show that a daily 1-hr dark pulse entrained the rhythm of wheel-running activity of unsplit hamsters. In addition, in split animals, unimodal coupling of the two locomotor activity components was produced by daily 1-hr dark pulses. In the SCN, the effects of entrainment and unimodal coupling of the two separate components by dark observed in behavior were also evident in the bilateral expression of the proteins c-FOS, p-ERK, PERIOD 1, and calbindin. These results show that the bilaterally asymmetric SCN clock, underlying split circadian behavior, can be recoupled in phase and entrained by short daily dark exposure, indicating the synchronizing potency of darkness on the main circadian clock.

A peripheral pacemaker drives the circadian rhythm of synaptic boutons in Drosophila independently of synaptic activity

Circadian rhythms in the morphology of neurons have been demonstrated in the fly Drosophila melanogaster. One such rhythm is characterized by changes in the size of synaptic boutons of an identified flight motor neuron, with larger boutons during the day compared with those at night. A more detailed temporal resolution of this rhythm shows here that boutons grow at a time of increased locomotor activity during the morning but become gradually smaller during the day and second period of increased locomotor activity in the evening. We have experimentally manipulated the synaptic activity of the fly during short periods of the day to investigate whether changes in bouton size might be a consequence of the different levels of synaptic activity associated with the locomotion rhythm of the fly. In the late night and early morning, when the flies normally have an intense period of locomotion, the boutons grow independently of whether the flies are active or completely paralyzed. Bouton size is not affected by sleep-deprivation during the early night. The cycle in bouton size persists for 2 days even in decapitated flies, which do not move, reinforcing the notion that it is largely independent of synaptic activity, and showing that a pacemaker other than the main biological clock can drive it.

Dysfonctionnements de l’horloge biologique et leurs traitements

Biological rhythms are periodic phenomena entrained to environmental changes by exogenous factors called synchronizers or entraining agents namely the light-dark cycle, the rest-activity cycle and the seasons, among others. In humans the major synchronizers are the light-dark and rest activity cycles. The endogenous component of a biological rhythm is dependent upon a number of clock genes. The main biological clock (oscillator or pacemaker) is the suprachiasmatic nucleus (SCN) of the anterior hypothalamus. The photoperiod (light-dark cycle) perceived by the retina acts on the SCN genes. Peripheral clocks have also been described in a number of tissues e.g. retina, adrenals. In a number of occurrences the synchronizers are badly perceived (transmeridian flights, shiftwork, nightwork...) or are not at all perceived (blindness). This situation is named rhythm desynchronization, it is external when the desynchronization is strictly related to the environment or internal when it is related to a dysfunction of the clock like in e.g. aging, Alzheimer disease, seasonal affective disorders (SAD) or hormone-dependent cancers which results in fatigue, sleep and mood disorders... A number of drugs called resynchronizing agents or chronobiotics which act on the biological clock are able to resynchronize the clock and to improve the patients' condition. Bright light is used in the treatment of SAD, melatonin, the pineal hormone, is also of interest when administered at precise timings in the 24hours scale. Other drugs like B12 vitamin or psychotropic drugs have also been proposed as chronobiotics.

Low reproductive success in Per1 and Per2 mutant mouse females due to accelerated ageing?

Recent studies on mice with mutations in the Clock gene have shown that this mutation disrupts oestrus cyclicity and interferes with successful pregnancy. In order to determine whether two other molecular components of the main clock, namely the period genes, Per1 and Per2, have an effect on the length of the oestrous cycle and the reproductive success, we used Per1- and Per2-deficient females. We show that although fecundity of young adult Per mutant females does not differ from that of wild-type females, middle-aged Per mutant mice are characterised by lower reproductive success than the control group. This may be a consequence of irregularity and acyclicity of the oestrous cycle of the middle-aged mutants. Besides, we demonstrate that Per mutant females have significantly more embryonal implantations in the uterus than successfully delivered offspring. The reproductive deficits of the middle-aged Per mutant females are comparable with those seen in aged wild-type mice. This suggests that Per1 and Per2 mutations cause an advanced ageing.

Serotonergic activation potentiates light resetting of the main circadian clock and alters clock gene expression in a diurnal rodent

The main circadian clock, localized in the suprachiasmatic nuclei (SCN) in mammals, can be synchronized by light and non-photic factors such as serotonergic cues. In nocturnal rodents, injections during the subjective day of the 5-HT 1A/7 receptor agonist 8-hydroxy-2-(di- n-propylamino) tetralin (8-OH-DPAT) or its positive enantiomer, induce behavioral phase-advances in correlation with decreased expression of two clock genes, Per1/2. In addition, 8-OH-DPAT and the selective serotonin reuptake inhibitor fluoxetine reduce light-induced phase-shifts during the subjective night. Beside the chronobiotic effects of serotonin, changes of serotonergic activity in humans have been involved in mood disorders, that are often associated with alterations in circadian rhythmicity. To get insights into the circadian role of serotonin in diurnal species, we investigated its modulation of the SCN in Arvicanthis ansorgei housed in constant darkness. In striking contrast to nocturnal rodents, daily serotonin content in Arvicanthis SCN peaked during daytime while the sensitivity window of its SCN to (+)8-OH-DPAT occurred essentially during the subjective night. Moreover, fluoxetine produced behavioral phase-advances at circadian time (CT) 0 and CT12. Expression of Per1/ 2, Rev-erbα/ β and Rorα/ β in the SCN was not modified after fluoxetine or (+)8-OH-DPAT injection. Furthermore, both treatments enhanced light-induced phase-advances and delays. Light responses of Per1 and Rorβ expression at CT0 and those of Per2 and Rev-erbα at CT12 were markedly altered by serotonergic activation. The present findings demonstrate that the serotonergic modulation of the SCN clock appears to differ between nocturnal species and the diurnal Arvicanthis. The potentiating effects of fluoxetine on light resetting in a diurnal rodent may be clinically relevant.