A Longitudinal Assessment of Sleep Timing, Circadian Phase, and Phase Angle of Entrainment across Human Adolescence

The aim of this descriptive analysis was to examine sleep timing, circadian phase, and phase angle of entrainment across adolescence in a longitudinal study design. Ninety-four adolescents participated; 38 (21 boys) were 9-10 years ("younger cohort") and 56 (30 boys) were 15-16 years ("older cohort") at the baseline assessment. Participants completed a baseline and then follow-up assessments approximately every six months for 2.5 years. At each assessment, participants wore a wrist actigraph for at least one week at home to measure self-selected sleep timing before salivary dim light melatonin onset (DLMO) phase - a marker of the circadian timing system - was measured in the laboratory. Weekday and weekend sleep onset and offset and weekend-weekday differences were derived from actigraphy. Phase angles were the time durations from DLMO to weekday sleep onset and offset times. Each cohort showed later sleep onset (weekend and weekday), later weekend sleep offset, and later DLMO with age. Weekday sleep offset shifted earlier with age in the younger cohort and later in the older cohort after age 17. Weekend-weekday sleep offset differences increased with age in the younger cohort and decreased in the older cohort after age 17. DLMO to sleep offset phase angle narrowed with age in the younger cohort and became broader in the older cohort. The older cohort had a wider sleep onset phase angle compared to the younger cohort; however, an age-related phase angle increase was seen in the younger cohort only. Individual differences were seen in these developmental trajectories. This descriptive study indicated that circadian phase and self-selected sleep delayed across adolescence, though school-day sleep offset advanced until no longer in high school, whereupon offset was later. Phase angle changes are described as an interaction of developmental changes in sleep regulation interacting with psychosocial factors (e.g., bedtime autonomy)

Timing system of HIRFL-CSR

The national science project HIRFL-CSR has recently been officially accepted. As a cyclotron and synchotron complex, it puts some particularly high demands on the control system. There are hundreds of pieces of equipment that need to be synchronized. An integrated timing control system is built to meet these demands. The output rate and the accuracy of the controller are 16 bit/mu s. The accuracy of the time delay reaches 40 ns. The timing control system is based on a typical event distribution system, which adopts the new event generation and the distribution scheme. The scheme of the tuning control system with innovation points, the architecture and the implemented method are presented in the paper.

Calcium-binding proteins in the circadian centers of the common marmoset (Callithrix jacchus) and the rock cavy (Kerodon rupestris) brains

The hypothalamic suprachiasmatic nucleus (SCN) and the thalamic intergeniculate leaflet (IGL) are considered to be the main centers of the mammalian circadian timing system. In primates, the IGL is included as part of the pregeniculate nucleus (PGN), a cell group located mediodorsally to the dorsal lateral geniculate nucleus. This work was carried out to comparatively evaluate the immunohistochemical expression of the calcium-binding proteins calbindin D-28k (CB), parvalbumin (PV), and calretinin (CR) into the circadian brain districts of the common marmoset and the rock cavy. In both species, although no fibers, terminals or perikarya showed PV-immunoreaction (IR) into the SCN, CB-IR perikarya labeling was detected throughout the SCN rostrocaudal extent, Seeming to delimit its cytoarchitectonic borders. CR-IR perikarya and neuropil were noticed into the ventral and dorsal portions of the SCN, lacking immunoreactivity in the central core of the marmoset and filling the entire nucleus in the rockcavy. The PGN of the marmoset presented a significant number of CB-, PV-, and CR-IR perikarya throughout the nucleus. The IGL of the rocky cavy exhibited a prominent CB- and CR-IR neuropil, showing similarity to the pattern found in other rodents. By comparing with literature data from other mammals, the results of the present study suggest that CB, PV, and CR are differentially distributed into the SCN and IGL among species. They may act either in concert or in a complementary manner in the SCN and IGL, so as to participate in specific aspects of the circadian regulation. (c) 2008 Elsevier Inc. All rights reserved.

Serotonin and circadian rhythms

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nucleotide binding and autophosphorylation of the clock protein KaiC as a circadian timing process of cyanobacteria

A negative feedback control of kaiC expression by KaiC protein has been proposed to generate a basic oscillation of the circadian clock in the cyanobacterium Synechococcus sp. PCC 7942. KaiC has two P loops or Walker's motif As, that are potential ATP-/GTP-binding motifs and DXXG motifs conserved in various GTP-binding proteins. Herein, we demonstrate that in vitro KaiC binds ATP and, with lower affinity, GTP. Point mutation by site-directed mutagenesis of P loop 1 completely nullified the circadian rhythm of kaiBC expression and markedly reduced ATP-binding activity. Moreover, KaiC can be autophosphorylated in vitro. These results suggest that the nucleotide-binding activity of KaiC plays important roles in the generation of circadian oscillation in cyanobacteria.

Discrete retinal input to the parabrachial complex of a new-world primate, the common marmoset (Callithrix jacchus)

Traditional retinal projections target three functionally complementary systems it) the brain of mammals: the primary visual system, the visuomotor integration systems and the circadian timing system. In recent years, studies in several animals have been conducted to investigate the retinal projections to these three systems, despite some evidence of additional targets. The aim of this study was to disclose a previously unknown connection between the retina and the parabrachial complex of the common marmoset, by means of the intraocular injection of cholera toxin Subunit b. A few labeled retinal fibers/terminals that are detected in the medial parabrachial portion of the marmoset brain show clear varicosities, Suggesting terminal fields. Although the possible role of these projections remains unknown, they may provide a modulation of the cholinergic parabrachial neurons which project to the thalamic dorsal lateral geniculate nucleus. (c) 2008 Elsevier Ireland Ltd. All rights reserved.

5-HT(1B) receptor in the suprachiasmatic nucleus of the common marmoset (Callithrix jacchus)

Serotonin (5-HT) is involved in the fine adjustments at several brain centers including the core of the mammal circadian timing system (CTS) and the hypothalamic suprachiasmatic nucleus (SCN). The SCN receives massive serotonergic projections from the midbrain raphe nuclei, whose inputs are described in rats as ramifying at its ventral portion overlapping the retinohypothalamic and geniculohypothalamic fibers. In the SCN, the 5-HT actions are reported as being primarily mediated by the 5-HT1 type receptor with noted emphasis for 5-HT(1B) subtype, supposedly modulating the retinal input in a presynaptic way. In this study in a New World primate species, the common marmoset (Callithrix jacchus), we showed the 5-HT(1B) receptor distribution at the dorsal SCN concurrent with a distinctive location of 5-HT-immunoreactive fibers. This finding addresses to a new discussion on the regulation and synchronization of the circadian rhythms in recent primates. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

Ritmo circadiano de atividade motora e distribuição diária dos comportamentos afiliativos ao longo da fase juvenil em saguis (callithrix jacchus)

The temporal allocation of the active phase in relation to light and dark cycle (LD) changes during puberty in humans, degus, rats and rhesus. In marmosets, the animal model used in several biomedical researches, there is evidence of a delay at the beginning of the active phase and an increase in total daily activity after onset of puberty. However, as this aspect was evaluated in animals maintained in natural environmental conditions, it was not possible to distinguish between the effects of puberty and of seasonality. Furthermore, as motor activity is the result of different behaviors in this species, it is also important to characterize the diurnal distribution of other behaviors in juvenile stage. With the aim of characterizing the circadian rhythm of motor activity and the diurnal profile of affiliative behavior in marmosets, the motor activity of 5 dyads juveniles between 4 and 12 months of age and their parents was recorded continuously for actímetro. The families were maintained under artificial LD 12:12 h, constant temperature and humidity. The duration of grooming behavior, proximity and social play among juveniles was recorded 2 times a week in sessions of 15 minutes each hour of the active phase. Afetr onset of puberty in juvenile, it was observed that there was no change in the parameters of circadian motor activity rhythm which were common to most animals. Despite the absence of pubertal modulation, it was observed that the circadian activity profiles have stronger synchrony between individuals of the same family than that of different families, which may indicate that the circadian activity rhythm was modulated by the dynamics of social interactions. In relation to age, the total daily activity and the ratio between evening and morning activity (EA/MA) were higher in juveniles than in adults, which may be associated with differences in the circadian timing system between age groups. Furthermore, the onset of the 10 consecutive hours of higher activity (M10) occurred earlier in adult males than in other members of the group, probably as a way to avoid competition for resources in one of the first activities of the day that is foraging. During the juvenile stage, there was an increase in total daily activity that may be associated with increased motor ability of juveniles. In addition to the circadian activity rhythm, the daytime profile of proximity and social play behaviors was similar between the 5th and 12th month of life of juveniles, in which the interval between 7- 10 h in the morning showed the highest values of proximity and lower values of play social. Moreover, the duration of the grooming showed a similar distribution to adults from the 8th month, wherein the higher values occurring at the interval between 11 14 h of day. Considering the results, the parameters of the circadian activity rhythm had a greater influence of social factors than puberty. In relation to age, there were no changes related to the allocation of the active phase in relation to the LD cycle, but total daily activity, the ratio AV/AM and the start of the M10 is possible to observe differences between juveniles and adults

Estudo da organização funcional do sistema circadiano por meio de ferramentas computacionais e matemáticas

Circadian rhythms are variations in physiological processes that help living beings to adapt to environmental cycles. These rhythms are generated and are synchronized to the dark light cycle through the suprachiasmatic nucleus. The integrity of circadian rhythmicity has great implication on human health. Currently it is known that disturbances in circadian rhythms are related to some problems of today such as obesity, propensity for certain types of cancer and mental disorders for example. The circadian rhythmicity can be studied through experiments with animal models and in humans directly. In this work we use computational models to gather experimental results from the literature and explain the results of our laboratory. Another focus of this study was to analyze data rhythms of activity and rest obtained experimentally. Here we made a review on the use of variables used to analyze these data and finally propose an update on how to calculate these variables. Our models were able to reproduce the main experimental results in the literature and provided explanations for the results of experiments performed in our laboratory. The new variables used to analyze the rhythm of activity and rest in humans were more efficient to describe the fragmentation and synchronization of this rhythm. Therefore, the work contributed improving existing tools for the study of circadian rhythms in mammals

Avaliação comportamental e neuroquímica de ratos em dessincronização forçada: possíveis implicações para um modelo animal de oscilações no humor

Bipolar disorder has been growing in several countries. It is a disease with high mortality and has been responsible by the social isolation of the patients. Bipolar patients have alterations in circadian timing system, showing a phase shift in various physiological variables. There are several arguments demonstrating alterations in circadian rhythms may be part of the bipolar disorder pathophysiology. Given the necessity for further elucidation, the goal of this study was to validate the forced desynchronization protocol as an animal model for bipolar disorder. To do this, Wistar rats were submitted to a forced desynchronization protocol which consists in a symmetrical light dark cycle with 22h. Under this protocol, rats dissociate the locomotor activity rhythm into two components: one synchronized to the light / dark cycle with 22h, and another component with period longer than 24 hours following the animal endogenous period. These rhythms with different periods sometimes there is coincidence, which we named CAP (Coincidence Active Phase) and the opposite phase, non-coincidence, called NCAP (Non-Concidence Active Phase). The hypothesis is that in CAP animals present a mania-like behavior and animals in NCAP depressive-like behavior. We found some evidence described in detail throughout this thesis. In sum, the animals under forced desynchronization protocol were more stressed, showed an increase in stereotypic behaviors such as grooming and reduction in other behaviors such as risk assessment and vertical exploration when compared to the control group. The CAP animals showed increased locomotor activity, especially during the dark phase when compared to controls (rats under T24) and less depressive behavior in the forced swim test. The animals in NCAP showed a higher anxiety in elevated plus maze, but they don t have ahnedonia. The animals under dissociation have more labeled 5HT1A cells at the amygdala area, which appoint that they have more amygdala inhibition. Taking these data together, we could partially validated the forced desynchronization protocol as an animal model for mood oscillations

Efeitos da senescência no núcleo supraquiasmático do sagui (Callithrix Jacchus): plasticidade morfológica e neuroquímica

The circadian timing system (CTS) is responsible for the generation and synchronization and the suprachiasmatic nucleus (SCN) of the hypothalamus has been described as the major circadian pacemaker in many mammalian species. The internal temporal organization managed by SCN is disturbed with aging bringing many pathological disorders that range from loss of complex cognitive performance to simple physiological functions. Therefore, our aim was perform a comparative study of the morphological aspects and neurochemical composition in the SCN of marmosets (Callithrix jacchus) adults and older using immunohistochemical techniques. We found morphometric and neurochemical changes in th SCN o folder animals in comparison to adults, among these a possible decreased in retinal projection to the SCN of older animals, found through a decline in CTB immunostaining, which can occur due atrophy and/or decreasing of fibers from the retinohypothalamic tract (RHT). The Klüver-Barrera histological technique strongly suggests a decrease in those fibers from RHT. Also, by means of a morphometric study, it is found a atrophy and numerical decline of neurons in SCN of aged animals, investigated by Nissl technique, and immunostaining with NeuN and calbindin. Relative optical density (ROD) analysis were used to evaluate the expression of some neurochemical components in SCN, such as GFAP expression, which was increased in older, result that indirectly reinforces that morphological changes occurs due the aging; the vasoactive intestinal polipeptide (VIP) showed no expression alteration in SCN of older animals; the serotonin (5-HT) was descreased in the dorsomedial portion of the SCN, and neurpeptide Y (NPY) apparently also decrease due to the increase of age. Many of these modifications were seen in other animals, such as rodents, human primates and non-human primates. These data about marmoset comes to add new information of the effect of aging on structures responsibles for the circadian rhytmicity, and that some behavioral changes controlled by th SCN, and founded in aged animals, may be caused by these morphological and neurochemical changes. Although some results have been quantitatively negative, qualitatively all analysis show significant change comparing adult and older animals, perhaps due to a low sampling number. In conclusion, the marmoset presents several morphological and neurochemical changes in the SCN of aged animals compared to adults, which may result in behavioral changes that favor pathology aging related

Efeitos de vocalizações de co-específicos e do escuro sobre o ritmo circadiano da atividade motora em sagüis (Callithrix jacchus)

The principal zeitgeber for most of species is the light-dark photocycle (LD), though other environment factors as food availability, temperature and social cues may act. Daily adjustment of the circadian pacemaker may result from integration of environmental photic and non-photic cues with homeostatic cues. Characterization of non-photic effects on circadian timing system in diurnal mammals is scarce in relation to nocturnal, especially for ecologically significant cues. Thus, we analyzed the effect of conspecific vocalizations and darkness on circadian activity rhythm (CAR) in the diurnal primate Callithirx jacchus. With this objective 7 male adults were isolated in a room with controlled illumination, temperature (26,8 ± 0,2°C) and humidity (81,6 ± 3,6%), and partial acoustic isolation. Initially they were under LD 12:12 (~300:2 lux), and subsequently under constant illumination (~2 lux). Two pulses of conspecific vocalizations were applied in total darkness, separated by 22 days, at 7:30 h (external time) during 1 h. They induced phase delays at circadian times (CTs) 1 and 10 and predominantly phase advances at CTs 9 and 15. After that, two dark pulses were applied, separated by 14 days, during 1 h at 7:30 h (external time). These pulses induced phase delays at CTs 2, 3 and 18, predominantly phase advances at CTs 8, 10 and 19, and no change at CT 14. However, marmosets CAR showed oscillations in endogenous period and active phase duration influenced by vocalizations from animals outside the experimental room, which interfered on the phase responses to pulses. Furthermore, social masking and relative coordination with colony were observed. Therefore, phase responses obtained in this work cannot be attributed only to pulses. Afterwards, pulses of conspecific vocalizations were applied in total darkness at 19:00 h (external time), during 1 h for 5 consecutive days, and after 21 days, for 30 consecutive days, on attempt to synchronize the CAR. No animal was synchronized by these daily pulses, although oscillations in endogenous period were observed for all. This result may be due to habituation. Other possibility is the absence of social significance of the vocalizations for the animals due to random reproduction, since each vocalization has a function that could be lost by a mixture of sounds. In conclusion, conspecific vocalizations induce social masking and relative coordination in marmosets CAR, acting as weak zeitgeber

Caracterização do ritmo de atividade motora durante a puberdade em sagüis (Callithrix jacchus) sob condições semi-naturais

Most of ontogenetic studies on circadian timing system have been developed on infants, adults and elderly. The puberty has not been a stage of life few studied, except for researches in human adolescents, that presents phase delay in sleep-wake cycle. However, few studies have focused on the basis of this circadian change due to methodological difficulties. Thus, an animal model to study the sleep-wake cycle at puberty is essential. In the common marmoset, a social primate, the circadian activity periodicity stabilizes around 4 months (juvenile stage) and the 8h period component has a seasonal variation. Puberty stage of this species begins near the 8th month of age in males and near the 7th month in females with 7 months of duration. With the aim to characterize the circadian motor activity rhythm during puberty in marmosets (Callithrix jacchus) the motor activity was continuous registered by actiwatches in 6 animals between 5-12 months. Since the social factor influence the behavior of this specie, behavioral observations were realized in 30 minutes windows twice/week to a general evaluation of the influence social interactions dynamic across experiment. Determination of puberty onset was done by fecal progesterone and estrogens in females, and androgens in males. From the analysis of the multiple regression test was selected a model that evaluate age and seasonal variables effect on the activity rhythm according to the higher explanation coefficient. The total activity was the only parameter influenced by age. Moreover, the activity onset was the parameter more explained by the model, and the sunrise was the factor that most influenced it. After the puberty onset, 2 dyads advanced the activity onset. The activity total decreased in 1 dyad and increased in 2 dyads. This increase may be related to the birth of infants in these families. The motor activity circadian component stabilized later in 1 dyad, coinciding with the puberty onset of these animals, while bimodality, caused by the 8 h component, was modulated by seasonality. The agonistic behavior was not evaluated due to reduced number of events. There were changes across ages in affiliative behavior of contact in 1 dyad, grooming done in 1 animal and grooming received in 2 animals. Although there is evidence of puberty effect on the activity motor rhythm, the photoperiodic fluctuations influenced the rhythm. Therefore is not possible to affirm if the puberty modulate the activity rhythm in marmosets

Perfil hormonal anual de machos e fêmeas adultos de saguis (callithrix jacchus)

Many behavioral and biological variables of animals are expressed in the form of biological rhytms, down by the Circadian Timing System, that synchronize them with the environment from external stimuli such as light. One of them is the secretion profile of most circulating hormones regulated by the hypothalamuspirtuitary axis, which controls functions essential for the survival and reproduction of organisms. The sagüi, Callithrix jacchus, one of the most studied species about their endocrine physiology, is an appropriate subject for evaluating the profile of plasma prolactin and cortisol of adult males and females born in captivity throughout the year. Three male and two adult femelas were housed individually and subjected to natural environmental conditions over two years. Blood samples were used to measure the circulating levels of both hormones by methods radioimmunoassay (RIA) and immunoassay (ELISA), respectively. The analysis during the year of the plasmatic values of both hormones test was performed by ANOVA for repeated measures, the correlation of Spearman, and the test of Friedman and Student's t-test. The levels of prolactin in plasma were higher during the months in which there is a greater incidence of births of baby in the colony, possibly serving for modulating the expression of the behavior of parental care in both sexes. The plasma cortisol showed a lift in anticipation of the station with the highest birth rate and may be associated with the preparation of individual participation in caring for the baby, and also with the establishment of emotional bond between reproductive partners. Thus, this study shows that, despite the variations observed in the environment in which the animals live, plasma levels of prolactin and cortisol vary little throughout the year.