蚤蝇的昼夜活动节律及睡眠行为研究

蚤蝇是重要的法医昆虫,同时是实验室中遗传、发育和生物测定等研究的重要对象。然而,蚤蝇的昼夜活动节律和睡眠行为及其在脑部的神经网络目前还不清晰。本文通过捕获本地蚤蝇并对其进行分子鉴定,研究了蚤蝇的昼夜活动节律和睡眠行为,同时表征了蚤蝇脑部核心钟神经元和多巴胺神经元。结果表明:蚤蝇在12h光照∶12h黑暗(12L∶12D)条件下不存在对开灯前或关灯前的活动预期,其双峰活动模式是对开关灯的光反应行为。在全黑暗(DD)条件下蚤蝇内源活动周期接近24h。黑腹果蝇神经肽PDF抗体免疫显示蚤蝇脑部核心钟神经元4~5个,不像黑腹果蝇一样存在明显的神经轴突。在睡眠行为上,蚤蝇雄虫和雌虫在整体活动强度、睡眠节律模式、总睡眠上均没有明显差异。相反,雄虫总睡眠次数和晚上睡眠次数低于雌虫,而总睡眠持续时间、晚上睡眠持续时间、总入睡时间和晚上入睡时间高于雌虫。此外,影响睡眠的重要多巴胺神经元在蚤蝇脑部的分布与黑腹果蝇类似。     

Rest-activity circadian rhythm and sleep quality in patients with binge eating disorder

Recent findings suggest that altered rest-activity circadian rhythms (RARs) are associated with a compromised health status. RARs abnormalities have been observed also in several pathological conditions, such as cardiovascular, neurological, and cancer diseases. Binge eating disorder (BED) is the most common eating disorder, with a prevalence of 3.5% in women and 2% in men. BED and its associate obesity and motor inactivity could induce RARs disruption and have negative consequences on health-related quality of life. However, the circadian RARs and sleep behavior in patients with BED has been so far assessed only by questionnaires. Therefore, the purpose of this study was to determine RARs and sleep parameters by actigraphy in patients with BED compared to a body mass index-matched control group (Ctrl). Sixteen participants (eight obese women with and eight obese women without BED diagnosis) were recruited to undergo 5-day monitoring period by actigraphy (MotionWatch 8®, CamNtech, Cambridge, UK) to evaluate RARs and sleep parameters. In order to determine the RARs, the actigraphic data were analyzed using the single cosinor method. The rhythmometric parameters of activity levels (MESOR, amplitude and acrophase) were then processed with the population mean cosinor.

The Actiwatch Sleep Analysis Software (Cambridge Neurotecnology, Cambridge, UK) evaluated the sleep patterns. In each participant, we considered seven sleep parameters (sleep onset: S-on; sleep offset: S-off; sleep duration: SD; sleep latency: SL; movement and fragmentation index: MFI; immobility time: IT; sleep efficiency: SE) calculated over a period of five nights.

The population mean cosinor applied to BED and Ctrl revealed the presence of a significant circadian rhythm in both groups (p < 0.001). The MESOR (170.0 vs 301.6 a.c., in BED and Ctrl, respectively; p < 0.01) and amplitude (157.66 vs 238.19 a.c., in BED and Ctrl, respectively p < 0.05) differed significantly between the two groups. Acrophase was not different between BED and Ctrl, as well as all sleep parameters. Both groups displayed a low level of sleep quality (SE 80.7% and 75.7% in BED and Ctrl, respectively). These data provided the first actigraphy-based evidence of RARs disruption and sleep behavior disorder in patients with BED. However, while sleep disorders could be reasonably ascribed to overweight/obesity and the related lower daily physical activity, RARs disruption in this pathology should be ascribed to factors other than reduced physical activity. The circadian timing approach can represent a novel potential tool in the treatment of patients with eating disorders. These data provide exploratory evidence of behavioral association in a small population of patients that, if confirmed in a wider number of subjects and across different populations, may lead to a revision and enhancement of interventions in BED patients.     

Circadian rhythm of autophagy proteins in hippocampus is blunted by sleep fragmentation

Autophagy is essential for normal cellular survival and activity. Circadian rhythms of autophagy have been studied in several peripheral organs but not yet reported in the brain. Here, we measured the circadian rhythm of autophagy-related proteins in mouse hippocampus and tested the effect of sleep fragmentation (SF). Expressions of the autophagy-related proteins microtubule‐associated protein 1 light chain 3 (LC3) and beclin were determined by western blotting and immunohistochemistry. Both the hippocampal LC3 signal and the ratio of its lipid-conjugated form LC3-II to its cytosolic form LC3-I showed a 24 h rhythm. The peak was seen at ZT6 (1 pm) and the nadir at ZT16 (1 am). The LC3 immunoreactivity in hippocampal CA1 pyramidal neurons also distributed differently, with more diffuse cytoplasmic appearance at ZT16. Chronic SF had a mild effect to disrupt the 24 h rhythm of LC3 and beclin expression. Interestingly, a greater effect of SF was seen after 24 h of recovery sleep when LC3-II expression was attenuated at both the peak and trough of circadian activities. Overall, the results show for the first time that the hippocampus has a distinct rhythm of autophagy that can be altered by SF.     

An investigation of circadian rhythm in Escherichia coli

Background: Persuasive evidence for circadian programs in non-photosynthetic bacteria other than cyanobacteria is still lacking, we aimed to investigate the circadian rhythm of specific growth rate in Escherichia coli ATCC 25922, one of the important prokaryotes. Methods: To grow E. coli under different light and dark conditions. When the growth entered into the stationary phase, we stopped the culture and obtained the viable counts by MTT assay every 3 h. The specific growth rates (SGRs) were calculated and analyzed with cosinor method for potential rhythms. Results: Single cosinor method revealed that the SGR of E. coli displayed rhythmic variations with a period of around 24 h both under light/dark cycles and under constant darkness. The best-fitting periods and best-fitting cosine curves were acquired. Conclusions: The SGR of E. coli (ATCC 25922) in a culture medium with limiting substrates in the stationary and death phases displayed rhythmic variations with a period of around 24 h under light/dark cycles and constant darkness conditions.     

Simulated microgravity influences circadian rhythm of NIH3T3 cells

Gravity heavily influences living organisms on earth including their circadian rhythm, which is fundamentally important for coordinately physiology in organisms as diverse as cyanobacteria, fungus and humans. Numerous researches have revealed that microgravity in outer space can affect circadian rhythm of astronauts and rodent animals, but the mechanism remains unknown. Using rotary cell culture system to simulate microgravity environment, we investigated the role of simulated microgravity in regulating the circadian rhythm of NIH3T3 cells. Our experiments found that simulated microgravity can not only influence the mRNA level of some core circadian genes, but also modify the circadian rhythm of Per1 and Per2 synchronized after phorbol myristate acetate treatment. Remarkably, MEK/ERK pathway was transiently activated after a 2-h simulated microgravity treatment, with a significant upregulation of Kras, Raf1 and p-ERK1/ERK2. Moreover, U0126, a selective inhibitor of MEK/ERK pathway, could disrupt the circadian rhythm of Per1 and Per2 synchronized after simulated microgravity treatment. Together, our results unveil that simulated microgravity could act like a zeitgeber to influence the circadian rhythm of NIH3T3 by acting on MEK/ERK pathway, indicating that MEK/ERK pathway may act as a bridge which connects cells mechanotransduction pathway and circadian rhythm regulation.     

Circadian rhythm of the locomotor activity in Drosophila melanogaster and its mutants 'sine oculis' and 'small optic lobes'

ABSTRACT. The locomotor activity patterns of wildtype Drosophila melanogaster and the mutants so (sine oculis) and sol (small optic lobes) were investigated. In all strains the proportions showing circadian rhythmicity, arrhythmicity and more complex patterns which could not be thus classified were similar. The occurrence of abnormal activity patterns is thus not a property of the mutation as previously claimed (Mack & Engelmann, 1981). In flies with a distinct circadian rhythmicity, the period lengths (τ) varied between strains, τ for wildtype Italy and the mutant so was longer than for wildtype Berlin and the mutant sol. As different τ's have been reported by others, τ does not seem to be closely determined for Drosophila melanogaster. Many flies exhibited two rhythms simultaneously, one with τ shorter and one with τ longer than 24 h, apparently implying two-oscillator control of the locomotor activity. The eyeless so mutants were entrained by LD cycles, so the compound eyes are not necessary, and so must possess the relevant photoreceptor(s) elsewhere. This may therefore also be true for the wildtype. Histology of the so adults revealed no correlation between the degree of reduction in the medulla and the occurrence of abnormal activity patterns. Since the only structures common to the medulla of so and sol are known to be large tangential cells, it is concluded that either they are of importance for the rhythmic system, or the oscillator(s) controlling locomotor activity is (are) not located in the medulla.     

Progressive retinal ganglion cell loss in primary open-angle glaucoma is associated with temperature circadian rhythm phase delay and compromised sleep

Advanced primary open-angle glaucoma (POAG) is characterized by progressive retinal ganglion cell complex (RGCC) damage that may cause subsequent disruption of the circadian rhythms. Therefore, we evaluated circadian body temperature (BT) rhythm and sleep characteristics of 115 individuals (38 men and 77 women) diagnosed with POAG. GLV (global loss volume; %), a measure of RGCC damage, was estimated by high-definition optical coherence tomography, and RGC functional ability was assessed by pattern electroretinogram amplitude (PERGA). Depending on dynamics of POAG progression criteria, two groups were formed that were distinctively different in GLV: Stable POAG group (S-POAG; GLV = 5.95 ± 1.84, n = 65) and Progressive POAG group (P-POAG; GLV = 24.27 ± 5.09, n = 50). S-POAG and P-POAG groups were not different in mean age (67.61 ± 7.56 versus 69.98 ± 8.15) or body mass index (24.66 ± 3.03 versus 24.77 ± 2.90). All subjects performed 21 around-the-clock BT self-measurements during a 72-h period and kept activity/sleep diaries. Results showed pronounced disruption of circadian physiology in POAG and its progression with increasing severity of the disease. The daily mean of BT was unusually low, compared to age-matched controls. Moreover, our results revealed distinctive features of BT circadian rhythm alterations in POAG development and POAG progression. S-POAG is associated with lowered BT circadian rhythm robustness and inter-daily phase stability compared to controls. In the P-POAG group, the mean phase of the circadian BT rhythm was delayed by about 5 h and phases were highly scattered among individual patients, which led to reduced group mean amplitude. Circadian amplitudes of individuals were not different between the groups. Altogether, these results suggest that the body clock still works in POAG patients, but its entrainment to the 24-h environment is compromised. Probably because of the internal desynchronization, bedtime is delayed, and sleep duration is accordingly shortened by about 55 min in P-POAG compared to S-POAG patients. In the entire POAG cohort (both groups), later sleep phase and shorter mean sleep duration correlate with the delayed BT phase (r = 0.215; p = 0.021 and r = 0.322; p = 0.0004, respectively). An RGCC GLV of 15% apparently constitutes a threshold above which a delay of the circadian BT rhythm and a shortening of sleep duration occur.     

Is there a 24-hour rhythm in alcohol craving and does it vary by sleep/circadian timing?

ABSTRACT

Increasing evidence implicates sleep/circadian factors in alcohol use; however, the role of such factors in alcohol craving has received scant attention. Prior research suggests a 24-hour rhythm in related processes (e.g., reward motivation), but more research directly investigating a rhythm in craving is needed. Moreover, prior evidence is ambiguous whether such a rhythm in alcohol craving may vary by sleep/circadian timing. To examine these possibilities, 36 late adolescents (18–22 years of age; 61% female) with regular alcohol use but without a current alcohol use disorder were recruited to complete smartphone reports of alcohol craving intensity six times a day for two weeks. During these two weeks, participants wore wrist actigraphs and completed two in-lab assessments (on Thursday and Sunday) of dim light melatonin onset (DLMO). Average actigraphically derived midpoint of sleep on weekends and average DLMO were used as indicators of sleep and circadian timing, respectively. Multilevel cosinor analysis revealed a 24-hour rhythm in alcohol craving. Findings across the sleep and circadian timing variables converged to suggest that sleep/circadian timing moderated the 24-hour rhythm in alcohol craving. Specifically, people with later sleep/circadian timing had later timing of peak alcohol craving. These findings add to the growing evidence of potential circadian influences on reward-related phenomena and suggest that greater consideration of sleep and circadian influences on alcohol craving may be useful for understanding alcohol use patterns and advancing related interventions.     

The role of circadian rhythm on the pharmacokinetic of methotrexate in streptozotocin-induced diabetes mellitus rats

Chronopharmacokinetic studies have been conducted both in animals and humans. Anticancer agents are of great interest due to their narrow therapeutic range and large pharmacokinetic variability. It was reported that the pharmacokinetics of MTX showed a circadian rhythm in rats and humans. Since diabetes-induced physiological changes can affect pharmacokinetics of drugs, it was reported that MTX blood concentration in diabetic rats was higher than that of the control groups. The present study was designed to elucidate whether these diabetes-induced changes in pharmacokinetics occurred during the day and thus administered MTX at four different times in streptozotocin-induced diabetes mellitus (SIDM) rats. Blood samples were drawn at 5, 15, 30, and 60 min after IV infusion of MTX in both the SIDM and control groups. Control and SIDM Area under the concentration - time curve (AUC) values showed a significant circadian rhythm with a peak located in mid-dark phase at 14:00. Clearance values were significantly low at 14:00 in the diabetic group when compared to other periods and the control group. The MTX AUC was increased when treatment with dexamethasone was given to suppress the endogenous production of corticosterone in both control and SIDM rats. These results suggest that the extent of MTX pharmacokinetics varies with the time of day in the SIDM rats and these variations might be related to changes in corticosterone concentrations.     

Sleep quality and methylation status of core circadian rhythm genes among nurses and midwives

Poor sleep quality or sleep restriction is associated with sleepiness and concentration problems. Moreover, chronic sleep restriction may affect metabolism, hormone secretion patterns and inflammatory responses. Limited recent reports suggest a potential link between sleep deprivation and epigenetic effects such as changes in DNA methylation profiles. The aim of the present study was to assess the potential association between poor sleep quality or sleep duration and the levels of 5-methylcytosine in the promoter regions of PER1, PER2, PER3, BMAL1, CLOCK, CRY1 CRY2 and NPAS2 genes, taking into account rotating night work and chronotype as potential confounders or modifiers. A cross-sectional study was conducted on 710 nurses and midwives (347 working on rotating nights and 363 working only during the day) aged 40–60 years. Data from in-person interviews about sleep quality, chronotype and potential confounders were used. Sleep quality and chronotype were assessed using Pittsburgh Sleep Quality Questionnaire (PSQI) and Morningness–Eveningness Questionnaire (MEQ), respectively. Morning blood samples were collected. The methylation status of the circadian rhythm genes was determined via quantitative methylation-specific real-time PCR assays (qMSP) reactions using DNA samples derived from leucocytes. The proportional odds regression model was fitted to quantify the relationship between methylation index (MI) as the dependent variable and sleep quality or sleep duration as the explanatory variable. Analyses were carried out for the total population as well as for subgroups of women stratified by the current system of work (rotating night shift/day work) and chronotype (morning type/intermediate type/evening type). A potential modifying effect of the system of work or the chronotype was examined using the likelihood ratio test. No significant findings were observed in the total study population. Subgroup analyses revealed two statistically significant associations between a shorter sleep duration and 1) methylation level in PER2 among day workers, especially those with the morning chronotype (OR = 2.31, 95%CI:1.24–4.33), and 2) methylation level in CRY2 among subjects with the intermediate chronotype, particularly among day workers (OR = 0.52, 95%CI:0.28–0.96). The study results demonstrated a positive association between average sleep duration of less than 6 hours and the methylation level of PER2 among morning chronotype subjects, and an inverse association for CRY2 among intermediate chronotype subjects, but only among day workers. Both the system of work and the chronotype turned out to be important confounders and modifiers in a number of analyses, making it necessary to consider them as potential covariates in future research on sleep deficiency outcomes. Further studies are warranted to explore this under-investigated topic.     

Time estimation circadian rhythm in shift workers and diurnally active humans

The ability of shift workers to estimate timer intervals of short duration was examined. The study included 22 shift workers and 10 diurnally working control subjects. A circadian rhythm in time estimates was documented in control subjects, but it was found to bed disrupted in shift workers. Spectral analysis revealed frequency or circadian component in time estimates to be lower among the shift workers. Furthermore, an interesting relationship was marked between time estimates and oral temperature in 4 control subjects and 6 shift workers in that the time of the closest estimation coincided with the peak time of their body temperature.     

The role of the epidermis in the generation of the circadian rhythm of carbon dioxide fixation in leaves of Bryophyllum fedtschenkoi

The role of the epidermis in the generation of the endogenous circadian rhythm of CO₂ exchange in leaves of Bryophyllum fedtschenkoi has been examined. At 25° C the rhythm of CO₂ output exhibited by whole leaves kept in continuous darkness and an initially CO₂-free air stream also occurs in isolated pieces of mesophyll. The sensitivity to light of the rhythms in whole leaves and in isolated mesophyll appears to be identical. At 15° C, however, no rhythm is observed in isolated mesophyll tissue, despite there being a conspicuous rhythm in intact leaves. The rhythm of net CO₂ assimilation in whole leaves kept in continuous light and a stream of normal air at either 25° C or at 15° C is abolished by removal of the epidermis, although at 15° C and under the higher of the two light levels used, there is an indication that rhythmicity may begin to reappear after the third day of the experiment. Thus, only under certain environmental conditions is the rhythm of CO₂ exchange in Bryophyllum leaves independent of the epidermis. The results indicate that the rhythm of carbon dioxide fixation in continuous darkness and CO₂-free air is generated primarily in the mesophyll cells, whereas the rhythm in continuous light and normal air is generated in the stomatal guard cells or in an interaction of these cells with the mesophyll cells.Abbreviation PEPCase phosphoenolpyruvate carboxylase     

Suprachiasmatic nucleus and circadian core temperature rhythm in the rat

Core temperature was telemetered from 26 singly-housed adult male inbred Fischer rats standardized in an ambient temperature of 24 ± 1°C, in light from 0600–1800 alternating with darkness (L:D 12:12), with food and water freely available. The rats were operated upon first for bilateral electrolytic lesioning of the suprachiasmatic nucleus (SCN) or by a sham-operation, which consisted of an inserted electrode which neither penetrated into the SCN area nor was activated to produce a lesion. Next, a temperature sensor was implanted intraperitoneally. The telemetered data obtained at 10-min intervals from each rat were analyzed by the least-squares fit of certain trial periods (cosinor methods). A circadian population rhythm persisted in the SCN-lesioned rats which sustained destruction of both SCN (P < 0.01). The amplitude of the circadian temperature rhythm was attenuated(P < 0.01) and the rhythm's acrophase advanced (P < 0.05) from mid-dark to a time near the transition from light to darkness. Unilateral lesions of the suprachiasmatic nuclei altered the circadain amplitude but not the phasing.     

Preoperative circadian physical activity rhythm and postoperative delirium in cardiovascular surgery patients

ABSTRACT

Postoperative delirium, a common perioperative complication, is frequently observed in elderly surgical patients. Few studies have investigated the life rhythm of preoperative patients, and whether or not the preoperative life rhythm is associated with the development of postoperative delirium. The purpose of this study was to investigate the relationship between the preoperative circadian physical activity rhythm and postoperative delirium in cardiovascular surgery patients. A total of 43 patients who underwent cardiovascular surgery were included in this prospective study between July 2016 and September 2017 at Kobe University Hospital. All subjects used a “Life Microscope” wristband (a wristwatch-type terminal incorporating a 3-axis accelerometer monitoring) for 3–7 days at home before the planned surgery. Hourly mean values were calculated for the metabolic equivalents from the obtained activity amounts, and subsequently evaluated using cosine periodic regression analysis. The circadian rhythm parameters of mesor (24 h time series mean), amplitude (half the peak-trough variation), and acrophase (peak time) for the metabolic equivalents were obtained. The intensive care delirium screening checklist was used to assess for postoperative delirium. The acrophase significantly advanced in the postoperative delirium group (median, 11:55 h [interquartile range, 11:06–12:27 h]) compared to the group without postoperative delirium (median, 13:25 h [interquartile range, 12:52–14:13 h]) (p < .001). Furthermore, binary logistic regression analysis showed that advances in the physical activity phase remained independently associated with postoperative delirium (odds ratio, 0.003 [95% confidence interval, 0–0.63]). These results suggest that misalignment between advanced life rhythm before hospitalization and life rhythm after hospitalization might be associated with risk for developing postoperative delirium. Our results led us to speculate that adequate consideration of the patient’s life rhythm before hospitalization is needed to prevent postoperative delirium.     

Case study of circadian rhythm sleep disorder following haloperidol treatment: reversal by risperidone and melatonin

A patient with Gilles de la Tourette syndrome treated with haloperidol, ingested once daily after awakening from sleep, exhibited an irregular sleep-wake pattern with a free-running component of approximately 48 h. Transfer to risperidone, ingested once daily after awakening from sleep, was beneficial resulting in a sleep-wake cycle more synchronized at the appropriate phase to the external zeitgebers, and fewer nocturnal disturbances. The circadian sleep-wake schedule was fully synchronized when the patient had been subsequently treated with melatonin at 21:00h, before intended nocturnal sleep, in addition to risperidone in the morning. Restoration of the sleep-wake circadian pattern was accompanied by the patient's subjective report of significant improvement in his quality of life, social interactions, and occupational status. This observation suggests that circadian rhythm sleep disorders can be related to the typical neuroleptic haloperidol and restored by the atypical neuroleptic risperidone. Similar findings reported in patients suffering from other disorders support the hypothesis that the described disruption of the sleep-wake schedule is medication rather than illness-related. Therefore, it is very important to realize that circadian rhythm sleep disorders may be a side effect of neuroleptics.     

Cell signaling,receptors, electrical effects and therapy in circadian rhythm

Abstract

Circadian rhythm has been the object of much attention. This review addresses the aspects of cell signaling, receptors, therapy and electrical effects in a multifaceted fashion. The pineal gland, which produces the important hormones melatonin and serotonin, exerts a prominent influence, in addition to the supraschiasmatic nucleus. Many aspects involve free radicals which have played a widespread role in biochemistry.     

月经周期对女性睡眠-觉醒和静息-活动的影响

目前有关月经周期对睡眠影响的研究结果并不一致,而对月经周期中昼夜睡眠-觉醒及静息-活动节律尚缺乏系统性的研究.本研究旨在观察正常育龄期女性月经周期中睡眠-觉醒及静息-活动昼夜节律的变化.我们采用静息-活动监测仪(actigraphy)和睡眠日志,调查了12个自然生活状态下健康育龄期妇女在月经周期不同阶段,即行经期、围排卵期、黄体早期及黄体晚期中睡眠与活动节律的变化.结果显示,睡眠-觉醒节律参数在四期之间无统计学显著差异;而静息-活动节律方面,所有受试女性静息-活动节律的平均日周期长度为(24.01±0.29)h,并且四期之间无显著性差异.行经期日间稳定系数(interdaily stability,IS)比黄体早期显著增加(P＜0.05).黄体早期日间活动开始时间明显较黄体晚期提前(P＜0.05);黄体早期的活动峰值时相比围排卵期显著提前(P＜0.05).月经周期可以影响静息-活动昼夜节律时相.而总体静息-活动数量与质量未发生显著变化;健康育龄期妇女在月经周期的各阶段中睡眠-觉醒节律亦无明显变异.     

Effects of weight, temperature and behaviour on the circadian rhythm of salivary cortisol in growing pigs

In farm animals, salivary cortisol has become a widely used parameter for measuring stress responses. However, only few studies have dealt with basal levels of concentration of cortisol in pigs and its circadian rhythm. The aim of this study was to examine the effects of ambient temperature and thermoregulatory behaviour on the circadian rhythm of salivary cortisol levels in fattening pigs. Subjects were 30 fattening pigs of different weight (60 to 100 kg), kept in six groups in an uninsulated building in pens with partly slatted floors. Saliva samples were taken every 2 h over periods of 24 h at different ambient temperatures at two times in winter and four times in summer. Thermoregulatory behaviour was recorded in the same 24-h time periods. The effect of time of day, body weight, ambient temperature and behaviour on the cortisol level was analysed using a mixed-effects model. Two peaks of cortisol levels per day were found. This circadian pattern became more pronounced with increasing weight and on days where thermoregulatory behaviour was shown. Mean cortisol levels per day were affected by weight but not by thermoregulatory behaviour. From our data, we conclude that long-term variations in cortisol concentration may be influenced by increasing age and weight more than by the respective experimental situation. In assessing animal welfare, it seems more reliable to consider the circadian pattern of cortisol concentration instead of only one value per day.     

Association of depressive symptoms with habitual sleep duration and sleep timing in junior high school students

ABSTRACT

This study aims to investigate independent associations of habitual sleep durations and sleep timings on weekdays and weekends with depressive symptoms in adolescents who have classes in the morning. We studied grade 7–9 students (942 males and 940 females, aged 12–15 years), who had classes in the morning, at public junior high schools in Japan in a cross-sectional design. The students answered a self-report questionnaire, which covers habitual sleep durations, bedtimes and wake-up times on weekdays and weekends, and depressive symptoms. The Short Mood and Feelings Questionnaire (SMFQ) was used to determine the level of depressive symptoms. The relationship between the variables on sleep habits and the SMFQ score were studied using multivariate linear regression and generalized additive models (GAM), controlling for sex, age and school. Multivariate linear regression analysis revealed that sleep duration on weekdays and relative mid-sleep time on weekdays (i.e. mid-sleep time on weekdays – mid-sleep time on weekends) were independently significantly (p < .001) associated with the SMFQ score. GAM analysis also revealed that sleep duration on weekdays (a reverse J-shaped relationship) and the relative mid-sleep time on weekdays (a negative monotonic/linear relationship) were independently significantly (p < .001) associated with the SMFQ score. These associations were confirmed in both males and females when they were analyzed separately. These results suggest that sleep duration on weekdays and the relative mid-sleep time on weekdays may be independently associated with the level of depressive symptoms in junior high school students who have classes in the morning. These findings may have important implications for the development of novel strategies for preventing mental health problems in adolescents.