Circadian Amplitude Research Articles

The circadian rhythm of blood pressure is masked by postural changes

It is well known that blood pressure (BP) varies throughout the day with lower levels observed during nocturnal sleep. This diurnal rhythm is partly explained by the BP-lowering effect of sleep and by an endogenous circadian rhythm of BP with a nocturnal through. As BP is acutely responsive to postural changes, the present study aimed at clarifying how posture affects the circadian variation of BP. Eleven healthy subjects (10 men, 1 woman; mean age ¡À SD: 22.6 ¡À 3.4 years old) were studied individually in time isolation for 6 consecutive days. After 2 baseline days on their regular sleep schedule, subjects underwent a 72-h ultradian sleep–wake cycle (USW) procedure, which consists of 60-min wake periods in dim light ( < 10 lux) alternating with 60-min naps in darkness ( < 0.3 lux). Subjects remained in a semi- recumbent posture throughout the first 48 h of the USW procedure. During the last 24 h of the USW procedure, they slept in a supine position during naps and changed into a sitting and standing position at 5 and 10 min after lights on, respectively. They returned to a sitting then supine position at 22 and 56 min after light on, respectively. BP was measured 1, 6, 11, 16 and 21 min after lights on. Circadian phase was assessed from core body temperature (CBT) (4x/min) which minimum was determined by a dual-harmonic regression and assigned a circadian phase of 0 degree. BP measures were averaged per subject by 60-degree bins and analyzed using non-linear mixed models. As expected, postural changes affected BP levels. No BP change was observed throughout wake periods when subjects remained in a semi-recumbent posture ( P ¡Ý0.38). In comparison, during the last 24 h of the USW procedure, diastolic (DBP) and mean arterial BP (MAP) increased by 11.97 ¡À 2.11 and 10.11 ¡À 2.34 mmHg, respectively, when participants changed from the supine to the standing position (measured 1 and 21 min after lights on, respectively; P < 0.001). Moreover, we observed a significant circadian rhythm of systolic BP (SBP), DBP, and MAP ( P ¡Ü0.001) during wake periods without postural change, with an acrophase occurring 7.4 ¡À 0.2 h, 8.6 ¡À 0.3 h and 7.0 ¡À 0.3 h after subjects¡ ¯ regular wake time, respectively. In comparison, during the postural change segment of the USW procedure, none of these rhythms were of sufficient amplitude to be significant, even though average values were comparable. Our observations indicate that postural changes significantly mask the endogenous circadian rhythm of BP. These results have practical implications for the measurement and interpretation of BP data collected in field conditions. Even when sleep is controlled for, the determination of circadian phase and amplitude of BP in ambulatory conditions is of limited value due to the confounding effects of changes in activity levels and posture. This research was supported by an operating grant from the Canadian Institutes of Health Research (CIHR). P. Boudreau was supported by a fellowship from the Institut de recherche Robert-Sauv ¨ ∣ en sant ¨ ∣ et en s ¨ ∣ curit ¨ ∣ du travail (IRSST).

Helix-loop-helix Protein Id2 Stabilizes Mammalian Circadian Oscillation Under Constant Light Conditions

The mammalian circadian oscillator is composed of interacting positive and negative transcription events. The clock proteins PER1 and PER2 play essential roles in a negative limb of the feedback loop that generates the circadian rhythm in mammals. In addition, the proteins CLOCK and BMAL1 (also known as ARNTL) form a heterodimer that drives the Per genes via the E-box consensus sequences within their promoter regions. In the present study, we demonstrate that Id2 is involved in stabilization of the amplitudes of the circadian oscillations by suppressing transcriptional activation of clock genes Clock and Bmal1. Id2 shows dynamic oscillation in the SCN, with a peak in the late subjective night. Under constant dark conditions (DD), Id2(-/-) mice showed no apparent difference in locomotor activity, however, under constant light conditions (LL), Id2(-/-) mice exhibit aberrant locomotor activity, with lower circadian oscillation amplitudes, although the free running periods in Id2(-/-) mice show no differences from those in either wild type or heterozygous mice. Id2(-/-) animals also exhibit upregulation of Per1 in constant light, during both the subjective night and day. In wild type mice, Id2 is upregulated by constant light exposure during the subjective night. We propose that Id2 expression in the SCN contributes to maintenance of dynamic circadian oscillations.

Dynamic Circadian Modulation in a Biomathematical Model for the Effects of Sleep and Sleep Loss on Waking Neurobehavioral Performance

Recent experimental observations and theoretical advances have indicated that the homeostatic equilibrium for sleep/wake regulation--and thereby sensitivity to neurobehavioral impairment from sleep loss--is modulated by prior sleep/wake history. This phenomenon was predicted by a biomathematical model developed to explain changes in neurobehavioral performance across days in laboratory studies of total sleep deprivation and sustained sleep restriction. The present paper focuses on the dynamics of neurobehavioral performance within days in this biomathematical model of fatigue. Without increasing the number of model parameters, the model was updated by incorporating time-dependence in the amplitude of the circadian modulation of performance. The updated model was calibrated using a large dataset from three laboratory experiments on psychomotor vigilance test (PVT) performance, under conditions of sleep loss and circadian misalignment; and validated using another large dataset from three different laboratory experiments. The time-dependence of circadian amplitude resulted in improved goodness-of-fit in night shift schedules, nap sleep scenarios, and recovery from prior sleep loss. The updated model predicts that the homeostatic equilibrium for sleep/wake regulation--and thus sensitivity to sleep loss--depends not only on the duration but also on the circadian timing of prior sleep. This novel theoretical insight has important implications for predicting operator alertness during work schedules involving circadian misalignment such as night shift work.

Tumor-Size Dependent Circadian Endocrine Disturbances in Untreated Patients With Primary Prostate Cancer

The aim of this study was to analyse the circadian profiles of central and peripheral circulating hormones in patients with unoperated primary localized prostate cancer (PC) as well as in age-matched controls with benign prostatic hyperplasia (BPH) in order see whether tumor-specific endocrine disturbances may occur. The hormones analysed in serum included melatonin, prolactin (PRL), growth hormone (GH), thyroid-stimulating hormone (TSH), total thyroxine (T4), luteinizing hormone (LH), testosterone (T), and cortisol (Cor). Blood was collected at four-hourly intervals over one complete 24-h cycle from 19 patients with PC (68 ± 3 years; T1N0M0-T4NxM0) and 20 patients with BPH (69 ± 3 years; BPH+-BPH+++). The most prominent result was that the circadian amplitude of melatonin was drastically depleted in PC (-71%) compared to BPH, and patients with T2 as well as T3/4 showed extremely low melatonin leading to a loss of circadian rhythmicity. A similar trend existed for PRL where no circadian rhythms were detectable in PC with larger tumors. Loss of circadian rhythmicity was also observed for GH in PC, and TSH-concentrations were clearly depleted by 43 – 53% in patients with T2-and T3/4-tumors compared to patients with medium- and large-sized BPH although T4 was within normal limits. LH as well as T and Cor did not show any changes in PC compared to BPH. This indicates that the growth of PC leads to specific endocrine changes affecting melatonin, PRL, GH and TSH without influencing peripheral hormones (T4, T, Cor) as well as LH. The possible significance of these findings for the prognosis of patients with PC is discussed, as well as the potential underlying mechanisms focussing on the question why circulating melatonin may be diminished in PC and whether this could be functionally involved in the observed endocrine disturbances.

Daily rest-activity patterns in the bipolar phenotype: A controlled actigraphy study

This study assessed daily rest-activity patterns in euthymic, medication-naïve bipolar phenotype individuals. The Mood Disorder Questionnaire was used to identify 19 bipolar phenotype individuals and 21 controls. Participants wore an Actiwatch-L for 2 weeks to assess their sleep behaviour and circadian rest-activity rhythmicity. Bipolar phenotype individuals had increased movement during sleep, as assessed by the fragmentation index, greater activity levels during their least active 5 h (2 am–7 am), and lower circadian relative amplitude compared to controls. Higher activity levels during sleep affecting circadian amplitude in young adults with the bipolar phenotype may be associated with vulnerability for developing mood disorder.

An out-of-lab trial: a case example for the effect of intensive exercise on rhythms of human clock gene expression

BackgroundAlthough out-of-lab investigation of the human circadian clock at the clock gene expression level remains difficult, a recent method using hair follicle cells might be useful. While exercise may function as an entrainment cue for circadian rhythms, it remains unclear whether exercise affects human circadian clock gene expression.MethodsEfforts to observe apparent effects of exercise on clock gene expression require that several specific conditions be met: intense exercise should be habitually performed at a relatively uncommon time of day over an extended period; and any relative phase shift thereby observed should be validated by comparison of exercise and no-exercise periods. Wake-up and meal times should be kept almost constant over the experimental period. The present study was conducted using a professional fighter who met these strict criteria as subject. Facial hair samples were collected at 4-h intervals around the clock to ascertain rhythms of clock gene expression.ResultsDuring a period in which nighttime training (from 20:00 to 22:00) was habitually performed, circadian clock gene expression was phase-delayed by 2 to 4 h compared with that during a no-exercise period. Maximum level and circadian amplitude of clock gene expression were not affected by the nighttime training.ConclusionOur trial observations illustrate the possibility that heavy physical exercise might strongly affect the circadian phase of clock gene expression. Exercise might be therefore effective for the clinical care of circadian disorders. The results also suggest that athletes may require careful scheduling of heavy physical exercise to maintain normal circadian phase and ensure optimal athletic performance.

Diminished leptin signaling can alter circadian rhythm of metabolic activity and feeding

Leptin, a hormone mainly produced by fat cells, shows cell-specific effects to regulate feeding and metabolic activities. We propose that an important feature of metabolic dysregulation resulting in obesity is the loss of the circadian rhythm of biopotentials. This was tested in the pan-leptin receptor knockout (POKO) mice newly generated in our laboratory. In the POKO mice, leptin no longer induced pSTAT-3 signaling after intracerebroventricular injection. Three basic phenotypes were observed: the heterozygotes had similar weight and adiposity as the wild-type (WT) mice (>60% of the mice); the homozygotes were either fatter (∼30%), or rarely leaner (<5%) than the WT mice. By early adulthood, the POKO mice had higher average body weight and adiposity than their respective same-sex WT littermate controls, and this was consistent among different batches. The homozygote fat POKO showed significant reduction of midline estimating statistic of rhythm of circadian parameters, and shifts of ultradian rhythms. The blunted circadian rhythm of these extremely obese POKO mice was also seen in their physical inactivity, longer feeding bouts, and higher food intake. The extent of obesity correlated with the blunted circadian amplitude, accumulative metabolic and locomotor activities, and the severity of hyperphagia. This contrasts with the heterozygote POKO mice which showed little obesity and metabolic disturbance, and only subtle changes of the circadian rhythm of metabolic activity without alterations in feeding behavior. The results provide a novel aspect of leptin resistance, almost manifesting as an "all or none" phenomenon.

P75 Neurotrophin Receptor Is a Clock Gene That Regulates Oscillatory Components of Circadian and Metabolic Networks

The p75 neurotrophin receptor (p75(NTR)) is a member of the tumor necrosis factor receptor superfamily with a widespread pattern of expression in tissues such as the brain, liver, lung, and muscle. The mechanisms that regulate p75(NTR) transcription in the nervous system and its expression in other tissues remain largely unknown. Here we show that p75(NTR) is an oscillating gene regulated by the helix-loop-helix transcription factors CLOCK and BMAL1. The p75(NTR) promoter contains evolutionarily conserved noncanonical E-box enhancers. Deletion mutagenesis of the p75(NTR)-luciferase reporter identified the -1039 conserved E-box necessary for the regulation of p75(NTR) by CLOCK and BMAL1. Accordingly, gel-shift assays confirmed the binding of CLOCK and BMAL1 to the p75(NTR-)1039 E-box. Studies in mice revealed that p75(NTR) transcription oscillates during dark and light cycles not only in the suprachiasmatic nucleus (SCN), but also in peripheral tissues including the liver. Oscillation of p75(NTR) is disrupted in Clock-deficient and mutant mice, is E-box dependent, and is in phase with clock genes, such as Per1 and Per2. Intriguingly, p75(NTR) is required for circadian clock oscillation, since loss of p75(NTR) alters the circadian oscillation of clock genes in the SCN, liver, and fibroblasts. Consistent with this, Per2::Luc/p75(NTR-/-) liver explants showed reduced circadian oscillation amplitude compared with those of Per2::Luc/p75(NTR+/+). Moreover, deletion of p75(NTR) also alters the circadian oscillation of glucose and lipid homeostasis genes. Overall, our findings reveal that the transcriptional activation of p75(NTR) is under circadian regulation in the nervous system and peripheral tissues, and plays an important role in the maintenance of clock and metabolic gene oscillation.

MicroRNA-185 oscillation controls circadian amplitude of mouse Cryptochrome 1 via translational regulation

Mammalian circadian rhythm is observed not only at the suprachiasmatic nucleus, a master pacemaker, but also throughout the peripheral tissues. Investigation of the regulation of clock gene expression has mainly focused on transcriptional and posttranslational modifications, and little is known about the posttranscriptional regulation of these genes. In the present study, we investigate the role of microRNAs (miRNAs) in the posttranscriptional regulation of the 3'-untranslated region (UTR) of the mouse Cryptochrome 1 (mCry1) gene. Knockdown of Drosha, Dicer, or Argonaute2 increased mCry1-3'UTR reporter activity. The presence of the miRNA recognition element of mCry1 that is important for miR-185 binding decreased mCRY1 protein, but not mRNA, level. Cytoplasmic miR-185 levels were nearly antiphase to mCRY1 protein levels. Furthermore, miR-185 knockdown elevated the amplitude of mCRY1 protein oscillation. Our results suggest that miR-185 plays a role in the fine-tuned regulation of mCRY1 protein expression by controlling the rhythmicity of mCry1 mRNA translation.

Sleepiness phenomics: Modeling individual differences in subjective sleepiness profiles

The present study investigates individual differences in subjective sleepiness profiles during 36h of sustained wakefulness in a modified constant routine protocol. Twenty-three volunteers (11 females), aged between 18 and 47yrs (M age=30.41, SD=10.26) enrolled in the study. Subjective sleepiness ratings were collected every 2h by means of visual analogue scales. Circadian rhythmicity was assessed by means of salivary cortisol. Subjective sleepiness data were analyzed using functional principal component analysis (fPCA). Our results show that approximately 80% of the variance is accounted for by three functional components. The first component explains 50.28% of the variance and is characterized by a profile of exclusively positive loadings, representing vertical shifts from the mean sleepiness profile. Scores on this component are positively related to self-reported habitual sleep times and mean slow wave activity (SWA) during wake. Positive scores on the second component (18.40% of the variance) are characterized by a higher than average peak-to-trough amplitude in subjective sleepiness profiles. Participants with higher than average scores on this component show a significantly higher amplitude in salivary cortisol profiles as opposed to participants with lower than average scores. Participants with positive scores on the third component (10.09% of the variance) show higher than average levels of subjective sleepiness during morning hours, a buildup of wake effort occurring later and more afternoon sleepiness after sleep deprivation than negative scorers. Peak levels of salivary cortisol occur significantly later in these participants. Taken together, our results suggest that component 1 represents tonic differences in sleepiness profiles primarily related to mechanisms of sleep homeostasis, component 2 to circadian amplitude differences and component 3 to diurnal preference. However, since the components are additions to a mean profile, each of the three components is likely to correspond to a mixture of multiple physiological parameters, rather than to a single process. The approach shows interesting potential for (1) revealing unidentified physiological processes, (2) testing existing assumptions about regulatory mechanisms at the basis of interindividual variability in sleepiness profiles and (3) the specification of sleepiness phenotypes on a quantitative basis.

Toward a Personalized Chronotherapy of High Blood Pressure and a Circadian Overswing

Timing can greatly affect the response to a stimulus, including antihypertensive medications. Herein, we assess the response of 30 patients to losartan/hydrochlorothiazide (L/H), administered for at least 1 month at a given circadian stage to each patient, this stage being changed during consecutive spans to cover six treatment times from awakening to bedtime at approximately 3-hour intervals. At the end of each stage, each patient underwent a 7-day around-the-clock ambulatory blood pressure (BP) profile, analyzed chronobiologically. A larger reduction of the midline estimating statistic of rhythm (MESOR; a rhythm-adjusted mean) of diastolic BP was achieved by L/H administration in the early morning for more patients (P < .05), while treatment upon awakening was the best choice for most patients to reduce the circadian amplitude of BP the most (P < .01). The optimal treatment time varied considerably among patients, however. Special attention should be given to the effect on the circadian amplitude since treatment can increase it above a threshold, beyond which there is a marked increase in cardiovascular disease risk. The results indicate the desirability to individualize the optimization of the antihypertensive effect of L/H by timing along the circadian scale.

Effects of the TRPV1 antagonist ABT‐102 on body temperature in healthy volunteers: pharmacokinetic/ pharmacodynamic analysis of three phase 1 trials

To characterize quantitatively the relationship between ABT-102, a potent and selective TRPV1 antagonist, exposure and its effects on body temperature in humans using a population pharmacokinetic/pharmacodynamic modelling approach. Serial pharmacokinetic and body temperature (oral or core) measurements from three double-blind, randomized, placebo-controlled studies [single dose (2, 6, 18, 30 and 40 mg, solution formulation), multiple dose (2, 4 and 8 mg twice daily for 7 days, solution formulation) and multiple-dose (1, 2 and 4 mg twice daily for 7 days, solid dispersion formulation)] were analyzed. NONMEM was used for model development and the model building steps were guided by pre-specified diagnostic and statistical criteria. The final model was qualified using non-parametric bootstrap and visual predictive check. The developed body temperature model included additive components of baseline, circadian rhythm (cosine function of time) and ABT-102 effect (Emax function of plasma concentration) with tolerance development (decrease in ABT-102 Emax over time). Type of body temperature measurement (oral vs. core) was included as a fixed effect on baseline, amplitude of circadian rhythm and residual error. The model estimates (95% bootstrap confidence interval) were: baseline oral body temperature, 36.3 (36.3, 36.4)°C; baseline core body temperature, 37.0 (37.0, 37.1)°C; oral circadian amplitude, 0.25 (0.22, 0.28)°C; core circadian amplitude, 0.31 (0.28, 0.34)°C; circadian phase shift, 7.6 (7.3, 7.9) h; ABT-102 Emax , 2.2 (1.9, 2.7)°C; ABT-102 EC50 , 20 (15, 28) ng ml(-1) ; tolerance T50 , 28 (20, 43) h. At exposures predicted to exert analgesic activity in humans, the effect of ABT-102 on body temperature is estimated to be 0.6 to 0.8°C. This effect attenuates within 2 to 3 days of dosing.

Association Between Phase Shifts, Expression Levels, and Amplitudes in Peripheral Circadian Clocks

Data analyses examining the relationship between circadian phase shifts and amplitudes are scarce. The aim of this data analysis was to explore the association between the phase shifts of gene expression, their amplitudes, and daily levels as a result of a given treatment under ad libitum or restricted feeding (RF) conditions. Two hundred forty data sets of gene expression (clock and metabolic genes) from various tissues and treatments were statistically analyzed. The data revealed a significant association between phase delays and increased amplitudes. Moreover, upon subgroup analyses, separating the RF from the non-RF groups, phase delays were significantly correlated with increased amplitudes and phase advances with decreased amplitudes. This picture was also achieved when clock genes, but not metabolic genes, were analyzed. In contrast, under RF, increased amplitude of metabolic genes correlated with phase advances. Moreover, phase advances under RF led to increased average daily levels in clock genes, but not in metabolic genes. In summary, these data demonstrated statistically significant association between phase shifts, daily levels, and amplitudes in circadian gene expression in peripheral tissues under timed versus ad libitum feeding conditions. (Author correspondence: oren.froy@mail.huji.ac.il)

Effects of insufficient sleep on circadian rhythmicity and expression amplitude of the human blood transcriptome

Insufficient sleep and circadian rhythm disruption are associated with negative health outcomes, including obesity, cardiovascular disease, and cognitive impairment, but the mechanisms involved remain largely unexplored. Twenty-six participants were exposed to 1 wk of insufficient sleep (sleep-restriction condition 5.70 h, SEM = 0.03 sleep per 24 h) and 1 wk of sufficient sleep (control condition 8.50 h sleep, SEM = 0.11). Immediately following each condition, 10 whole-blood RNA samples were collected from each participant, while controlling for the effects of light, activity, and food, during a period of total sleep deprivation. Transcriptome analysis revealed that 711 genes were up- or down-regulated by insufficient sleep. Insufficient sleep also reduced the number of genes with a circadian expression profile from 1,855 to 1,481, reduced the circadian amplitude of these genes, and led to an increase in the number of genes that responded to subsequent total sleep deprivation from 122 to 856. Genes affected by insufficient sleep were associated with circadian rhythms (PER1, PER2, PER3, CRY2, CLOCK, NR1D1, NR1D2, RORA, DEC1, CSNK1E), sleep homeostasis (IL6, STAT3, KCNV2, CAMK2D), oxidative stress (PRDX2, PRDX5), and metabolism (SLC2A3, SLC2A5, GHRL, ABCA1). Biological processes affected included chromatin modification, gene-expression regulation, macromolecular metabolism, and inflammatory, immune and stress responses. Thus, insufficient sleep affects the human blood transcriptome, disrupts its circadian regulation, and intensifies the effects of acute total sleep deprivation. The identified biological processes may be involved with the negative effects of sleep loss on health, and highlight the interrelatedness of sleep homeostasis, circadian rhythmicity, and metabolism.

Non-optimal codon usage is a mechanism to achieve circadian clock conditionality

Circadian rhythms are oscillations in biological processes that function as a key adaptation to the daily rhythms of most environments. In the model cyanobacterial circadian clock system, the core oscillator proteins are encoded by the gene cluster kaiABC1. Genes with high expression and functional importance like the kai genes are usually encoded by optimal codons, yet the codon usage bias of the kaiBC genes is not optimized for translational efficiency. We discovered a relationship between codon usage and a general property of circadian rhythms called conditionality; namely, that endogenous rhythmicity is robustly expressed under some environmental conditions but not under others2. Despite the generality of circadian conditionality, however, its molecular basis is unknown for any system. Here we show that non-optimal codon usage was selected as a post-transcriptional mechanism to switch between circadian and non-circadian regulation of gene expression as an adaptive response to environmental conditions. When the kaiBC sequence was experimentally optimized to enhance expression of the KaiB and KaiC proteins, intrinsic rhythmicity was enhanced at cool temperatures that are experienced by this organism in its natural habitat. However, fitness at those temperatures was highest in cells whose endogenous rhythms were suppressed at cool temperatures as compared with cells exhibiting high-amplitude rhythmicity. These results indicate natural selection against circadian systems in cyanobacteria that are intrinsically robust at cool temperatures. Modulation of circadian amplitude is therefore critical to its adaptive significance3. Moreover, these results show the direct effects of codon usage on a complex phenotype and organismal fitness. Our work also challenges the long-standing view of directional selection towards optimal codons4–7, and provides a key example of natural selection against optimal codon to achieve adaptive responses to environmental changes.

Nascent-Seq analysis of Drosophila cycling gene expression

Significance Statement Rhythmic mRNA expression is a hallmark of circadian rhythms. To address this regulation in Drosophila , we used high throughput sequencing to compare nascent RNA and mRNA levels in fly heads at different times of day. This indicated that some genes undergo rhythmic transcription with no impact on mRNA, whereas a few strongly rhythmic mRNAs have weak transcriptional oscillations. A substantial number of rhythmic genes manifest more robust mRNA cycling than transcriptional cycling, indicating that post-transcriptional regulation plays a widespread role in boosting circadian gene expression amplitude. Our study highlights the importance of directly assaying transcription to understand gene regulation.

Structural and Electrical Myocardial Remodeling in a Rodent Model of Depression

Despite a well-documented association between stress and depression with cardiac morbidity and mortality, there is no satisfactory explanation for the mechanisms linking affective and cardiac disorders. This study investigated cardiac electrophysiological properties in an animal model of depression. Depression-relevant physiological and behavioral parameters were measured in adult male wild-type rats during and after a period of intermittent social defeat stress (n = 12) or empty cage exposure (control, n = 11). Nine days after the last defeat/empty cage exposure, high-definition epicardial mapping was performed under anesthesia. Stressed animals versus controls displayed a larger reduction in the circadian amplitude of heart rate (-32% [3%] versus -13 [2%]; p = .001) and body temperature (-33% [4%] versus -5% [2%]; p = .001) rhythms, had smaller body weight gain (+11% [1%] versus +17% [1%]; p < .001), and showed a larger reduction in sucrose solution intake (-19% [6%] versus -7% [4%]; p = .006). Epicardial mapping analysis revealed a decrease in the transversal conduction velocity of the wavefront (0.23 [0.0] versus 0.27 [0.1] m/s; p = .02) and a shortening of the effective refractory period (86.8 [2.1] versus 95.9 [3.0] milliseconds; p = .01) in stressed animals. Upon killing, moderate left ventricular fibrosis was observed in the stressed group. Intermittent social stress procedure is associated with depression-like symptoms and altered myocardial electrical stability in a potentially proarrhythmic manner. In particular, reduced myocardial refractoriness and impaired conduction, which are considered major determinants of arrhythmogenesis, represent possible mechanisms underlying cardiac vulnerability.

Aging-like circadian disturbances in folate-deficient mice

The elderly population shows various circadian disturbances, including dampened amplitude of rhythmicity and decreased responsiveness to light. The common poor folate status in the elderly might account for these aging-related circadian disturbances. To test this hypothesis, we investigated whether folate deficiency in mice affects circadian oscillations of the master clock in the suprachiasmatic nuclei, and the shifting responses to light. Mice fed a diet without folate for 6 weeks displayed markedly reduced (4.5-fold) erythrocyte folate concentration and increased (2.3-fold) homocysteinemia compared with control mice. Folate deficiency decreased the circadian amplitude of vasopressin and the clock protein PERIOD 2 (PER2) in the master clock, slowed the rate of re-entrainment of behavioral rhythms after delayed light-dark cycle and reduced light-induced phase-delays, without detectable morphologic changes in the retina, such as the number of melanopsinergic ganglion cells, that might have impaired photodetection. In conclusion, folate deficiency and consecutive hyperhomocysteinemia led to dampened PER2 and vasopressin oscillations in the master clock and reduced responsiveness to photic resetting, which constitute hallmarks of aging effects on circadian rhythmicity.

Circadian amplitude and homeostatic buildup rate in postural control

Postural control during quiet stance is a common everyday physical activity. Sleepiness is increasingly prevalent in our 24-h society. Yet, little research exists that quantitatively links the fluctuations in sleepiness and postural control. This study quantifies the circadian amplitude and homeostatic buildup rate in postural control. With a force plate we assessed postural control in 12 participants (21–38 years) every 2h during 24h of sustained wakefulness. The sway area was 1.39±0.71mm2 at the circadian high around noon, and 4.02±0.67mm2 at the circadian low around 6 am (a 189% change, p=0.02). The circadian amplitude of the sway area was therefore 2.63mm2. The sway area was 1.92±0.64mm2 at the start of the 24-h period and 4.42±0.69mm2 at the end of the period (a 130% change, p<0.001). The homeostatic buildup rate of sway area was 0.04h−1. The circadian- and homeostatic effects on sway variability, sway velocity, sway frequency and fractal dimension were smaller but still significant. This study found that the circadian amplitude and homeostatic buildup rate are quantifiable from posturographic data, and that they have significant impact on postural control. This finding is important because it means that one could apply the framework of the famous two-process model of sleep regulation (published by Borbély in 1982) to explain the previously reported sleepiness-related changes in postural control.

Mis-expression of the BK K+ channel disrupts suprachiasmatic nucleus circuit rhythmicity and alters clock-controlled behavior

In mammals, almost all aspects of circadian rhythmicity are attributed to activity in a discrete neural circuit of the hypothalamus, the suprachiasmatic nucleus (SCN). A 24-h rhythm in spontaneous firing is the fundamental neural intermediary to circadian behavior, but the ionic mechanisms that pattern circuit rhythmicity, and the integrated impact on behavior, are not well studied. Here, we demonstrate that daily modulation of a major component of the nighttime-phased suppressive K(+) current, encoded by the BK Ca(2+)-activated K(+) current channel (K(Ca)1.1 or Kcnma1), is a critical arbiter of circadian rhythmicity in the SCN circuit. Aberrant induction of BK current during the day in transgenic mice using a Per1 promoter (Tg-BK(R207Q)) reduced SCN firing or silenced neurons, decreasing the circadian amplitude of the ensemble circuit rhythm. Changes in cellular and circuit excitability in Tg-BK(R207Q) SCNs were correlated with elongated behavioral active periods and enhanced responses to phase-shifting stimuli. Unexpectedly, despite the severe reduction in circuit amplitude, circadian behavioral amplitudes in Tg-BK(R207Q) mice were relatively normal. These data demonstrate that downregulation of the BK current during the day is essential for the high amplitude neural activity pattern in the SCN that restricts locomotor activity to the appropriate phase and maintains the clock's robustness against perturbation. However, a residually rhythmic subset prevails over the ensemble circuit to drive the fundamental circadian behavioral rhythm.