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.

Efeito do exercício físico matinal realizado sob luz solar no ciclo vigília-sono de adolescentes

The sleep onset and offset delay at adolescence in relation to childhood. Besides biological causes, some external factors as academic obligations and socialization contributes, increasing the burden of school and socialization. However, morning school schedules reduce sleep duration. Besides light strong effect, studies in humans have indicated that exercise influence circadian synchronization. To evaluate the effect of the morning exercise under sunlight on sleep-wake cycle (SWC) of adolescents, 160 high school students (11th year) were exposed to the following conditions: lesson in usual classroom (Group C), lesson in swimming pool exposed to sunlight (Group E), half of them carrying through physical activity (EE) and the other resting (EL). Each experimental group met two stages: assessment of SWC 1 week before and 1 week during the intervention, which was held in Monday and Wednesday between 7:45 and 8:30 am. In the baseline, there were applied the questionnaires "Health and Sleep" and cronotype evaluation (H & O). In addition, students were evaluated before and during the intervention by "Sleep Diary", "Karolinska Sleepiness Scale" (KSS), Psychomotor Vigilance Test (PVT) and actimetry. During the intervention, there was a delay in wake-up time on the weekend and a trend to greater sleep duration on week for the three groups. At the weekend, only the groups EE and EL increased sleep duration. There was no difference in bedtime, irregularity of sleep schedules and nap variables. The sleepiness showed a circadian pattern characterized by higher alertness levels at 11:30 am and sleepiness levels at bedtime and wake-up time on week. On weekends there were higher levels of alertness in these times. In the days of intervention, there was an increase of sleepiness at 11:30 am for groups EL and EE, which may have been caused by a relaxing effect of contact with the water of the pool. In addition, the group EE showed higher alert levels at 14:30 pm on Monday and at 8:30 am in the Wednesday, possibly caused by exercise arousal effect. The reaction time assessed through the TPV did not vary between the stages. The sleep quality improved in the three groups in the second stage, making impossible the evaluation of intervention effect. However, the sleep quality increased on Monday and Tuesday only on the group EE. From the results, it is suggested that the intervention promoted effects on the sleepiness at some day hours. In other SWC variables there were no effects, possibly due to a large SWC irregularity on weekends. Thus, the evaluation of higher weekly frequency EF is necessary, since only two days were insufficient to promote greater effect on adolescents SWC

A Zona Incerta no sagüi (Callithrix jacchus): Análise Citoarquitetônica, Neuroquímica e Projeção Retiniana

The retinal projections in mammals usually reach, classically, three major functional systems: the primary visual system, the accessory optic system, and the circadian timing system. But the retinal projections also reach areas classically considered non-visual, one of which groups the neurons of the zona incerta (ZI), target this study. The primary visual system includes thalamic lateral geniculate complex is formed by the dorsal lateral geniculate nucleus, intergeniculate leaflet and the ventral lateral geniculate nucleus and other Components. The accessory optic system is composed of the small nuclei: nuclei terminal dorsal, lateral, medial and the interstitial nucleus of the superior posterior fasciculus. These nuclei are involved in visuo-motor activities. The circadian timing system is comprised of the suprachiasmatic nucleus of the hypothalamus, that act as master circadian pacemaker, entraining pathways and efferents pathways to the efectors, and the intergeniculate leaflet, that seems to act as a modulator of the pacemaker. The retinal projections too reach classically considered non-visual areas, including the zona incerta. This region is localized in the ventral thalamus and has been implicated in various functional properties including nociceptive and somatosensory processing, motor response, sociosexual behaviour, feeding and drinking, in symptoms of neurodegenerative diseases, arousal and attention. It also displays connection with several areas of central nervous system. The aim of this study was characterize the retinal projection in the zona incerta of Callithrix jacchus (sagüi), a primate of the New World through the anterograde axonal transport of the cholera toxin subunit b and analyze the citoarchicteture using Nissl and NeuN, and neurochemical substances such as serotonin, GABA, VIP, VP, GFAP and binding-calcium proteins. The zona incerta showed a different division of the literature in citoarquitetura, both by means of Nissl as neurochemical by NeuN, with a subdivision ventrolateral and dorsomedial. The neurochemical to the other substances corroborate with this subdivision. The GFAP was almost completely negative for the zona incerta, result non evidenced in previous studies yet. The 16 retinal projection in sagüi, unlike other primates and rodents, reached the caudal portion only. This work helps to make further studies are conducted based on this subdivision and the localization of the neurochemical substances associated with possible behaviors that the zona incerta is involved

Influência de fotoperíodo artificial no comportamento de um primata neotropical diurno (Callithrix jacchus)

One of the main environmental cues for the adjustment of temporal organization of the animals is the light-dark cycle (LD), which undergoes changes in phase duration throughout the seasons. Photoperiod signaling by melatonin in mammals allows behavioral changes along the year, as in the activity-rest cycle, in mood states and in cognitive performance. The aim of this study was to investigate if common marmoset (Callithrix jacchus) exhibits behavioral changes under short and long photoperiods in a 24h cycle, assessing their individual behaviors, vocal repertoire, exploratory activity (EA), recognition memory (RM) and the circadian rhythm of locomotor activity (CRA). Eight adult marmosets were exposed to a light-dark cycle of 12:12; LD 08:16; LD 12:12 and LD 16:08, sequentially, for four weeks in each condition. Locomotor activity was recorded 24h/day by passive infrared motion detectors above the individual cages. A video camera system was programmed to record each animal, twice a week, on the first two light hours. From the videos, frequency of behaviors was registered as anxiety-like, grooming, alert, hanging position, staying in nest box and feeding using continuous focal animal sampling method. Simultaneously, the calls emitted in the experimental room were recorded by a single microphone centrally located and categorized as affiliative (whirr, chirp), contact (phee), long distance (loud shrill), agonistic (twitter) and alarm (tsik, seep, see). EA was assessed on the third hour after lights onset on the last week of each condition. In a first session, marmosets were exposed to one unfamiliar object during 15 min and 24h later, on the second session, a novel object was added to evaluate RM. Results showed that long days caused a decreased of amplitude and period variance of the CRA, but not short days. Short days decreased the total daily activity and active phase duration. On long days, active phase duration increased due to an advance of activity onset in relation to symmetric days. However, not all subjects started the activity earlier on long days. The activity offset was similar to symmetric days for the majority of marmosets. Results of EA showed that RM was not affected by short or long days, and that the marmosets exhibited a decreased in duration of EA on long days. Frequency and type of calls and frequency of anxiety-like behaviors, staying in nest box and grooming were lower on the first two light hours on long days. Considering the whole active phase of marmosets as we elucidate the results of vocalizations and behaviors, it is possible that these changes in the first two light hours are due to the shifting of temporal distribution of marmoset activities, since some animals did not advance the activity onset on long days. Consequently, the marmosets mean decreased because the sampling was not possible. In conclusion, marmosets synchronized the CRA to the tested photoperiods and as the phase angle varied a lot among marmosets it is suggested that they can use different strategies. Also, long days had an effect on activity-rest cycle and exploratory behaviors

Influência da auto seleção à luz no ritmo circadiano de atividade motora em Callithrix Jacchus

Marmosets, Callithrix jacchus, are strictly diurnal animals. The motor activity rhythmicity is generated by the circadian timing system and is modulated by environmental factors, mainly by photic stimuli that compose the light-dark cycle. Photic stimuli can reset the biological oscilators changing activity motor pattern, by a mechanism called entrainment. Otherwise, light can act directly on expressed rhythm, without act on the biological oscillators, promoting the masking. Thus, photic stimuli can synchronize the circadian activity rhythm (CAR) by two distinct mechanisms, acting isolated or at a combined way. Among the elements that can influence photic synchronization, the duration and time of photic exposure is pointed out. If in the natural environment the marmoset can choose places of different intensity illumination and is synchronized to light-dark cycle (LD), how the photic synchronization mechanism can be evaluated in laboratory by light self-selection? With objective to response this question, four adult male marmosets were studied at two conditions: with and without sleeping box. The animals were submitted to a LD cycle (12:12/ 350:2 lx) and constant light (LL: 350 lx) conditions in individual cages with an opaque sleeping box, that permitted the light self-selection. At the room, the temperature was 25.6 ºC (± 0.3 ºC) and humidity was 78.7 (± 5%). The motor activity was recorded at 5 min bins by infrared movement sensors installed at the top of the cages. The motor activity profile was distinct at the two conditions: without the sleeping box protection against light, the activity frequency was higher at CT 11-12 (ANOVA; F(3.23) = 62.27; p < 0.01). Also, the duration of the active phase (α) was prolonged of about 1 h (t test, p < 0.05) and the animals showed a significant delay on the activity onset and offset (t test, p < 0.05) and at the acrophase (confidence intervals of 5%) of CAR. In LL, the light continuous exposure prolonged the active phase and influenced the endogenous expression of the circadian activity rhythm period. From the result analysis, it is concluded that the light self-selection can modify several parameters of CAR in marmosets, allowing the study of the synchronization mechanism using the burrow model. Thus, without sleeping box there was a phase delay between the CAR and LD (entrainment) and an increase of activity near lights off (positive masking). Furthermore, in LL, the light continuous exposure modifies α and the endogenous expression of CAR. It is suggested that the light self-selection might be take into account at investigations that evaluate the biological rhythmicity in marmosets

Estampa temporal na memória de reconhecimento em estudantes universitários

Several studies have shown that there is a circadian modulation of explicit memory. This modulation can occur independently in each one of the mnemonic processes. The aim of this study was to evaluate the influence of time of training on short-term memory (STM) and long-term memory (LTM), using a recognition task. Moreover, a possible circadian modulation in retrieving was investigated when this process matched the acquisition hour (time stamp). The chronotype variable was also considered. Fifty-seven undergraduate students aging between 18 and 25 years (21,72 ± 2,14; 28 ♂) participated in this study. In the training phase (acquisition) the subjects heard a ten word list. Following this, they answered a recognition test to evaluate STM and one week later they answered a recognition test to evaluate LTM. In each chronotype, the subjects were divided in groups according to the training hour, part of them in the morning and the other in the afternoon. One week later some of the subjects in each group underwent LTM testing in the morning and others in the afternoon. When the subjects performances were analyzed together, independently of the chronotypes, a training hour effect was found in the LTM. The subjects trained in the afternoon had better performance. No time of day effect was found in the STM and in retrieving from the LTM. However, the morning types who were trained and tested in the same hour had a better performance in the LTM when compared to morning types trained and tested in different hours. This effect did not occur when the other chronotypes were analyzed. The circadian modulation seems to occur at least in two different ways. First, there is a circadian modulation in the acquisition/consolidation processes, with a better performance occuring in the afternoon. Secondly, there is a modulation in the retrieval mnemonic process, called time stamp phenomenon. This phenomenon, that occurred in the morning types, is showed for the first time in humans

Time-dependent alterations of soluble and cellular components in human milk

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

Picos de pressão intra-ocular: comparação entre curva tensional diária, minicurva e medida da pressão intra-ocular às 6 horas

INTRODUÇÃO: A pressão intra-ocular (Po) é o fator de risco isolado mais importante para o desenvolvimento do glaucoma primário de ângulo aberto (GPAA). O controle da Po é o objetivo principal da terapia antiglaucomatosa até o momento. A curva tensional diária (CTD) é de grande importância para o diagnóstico e seguimento do glaucoma primário de ângulo aberto. Métodos simplificados como a minicurva têm sido utilizados em seu lugar por serem mais práticos. OBJETIVO: Comparar curva tensional diária, minicurva e medida isolada às 6 horas quanto à detecção de picos pressóricos e verificar a influência da variação postural na medida das 6 horas da manhã. MÉTODOS: Sessenta e quatro pacientes (126 olhos) com glaucoma primário de ângulo aberto ou suspeita de glaucoma foram submetidos à curva tensional diária. A minicurva considerou as medidas das 9, 12, 15 e 18 horas da mesma curva tensional diária. A medida das 6 horas foi realizada no escuro, com o paciente deitado, utilizando o tonômetro de Perkins. Logo após, foi feita nova medida, com o paciente sentado, usando o tonômetro de Goldmann. A Po média e a ocorrência de picos (Po > 21 mmHg) da curva tensional diária e minicurva foram comparados, assim como o horário de ocorrência dos picos. RESULTADOS: A Po média foi maior no glaucoma primário de ângulo aberto do que nos suspeitos tanto na minicurva como na curva tensional diária Quando comparadas, a curva tensional diária apresentou médias de Po maiores que a minicurva. A medida das 6 horas foi maior quando feita com o paciente deitado. A minicurva não detectou 60,42% dos picos nos pacientes com glaucoma primário de ângulo aberto e 88,24% dos picos nos suspeitos. CONCLUSÃO: 1. A curva tensional diária detectou mais picos pressóricos do que a minicurva; 2. A média de Po das 6 horas foi maior com o paciente deitado; 3. A Po das 6 horas com o paciente deitado foi maior do que a Po média da curva tensional diária e da minicurva.

Ritmo Circadiano de Triiodotironina (T3) e Tiroxina (T4) Plasmática em Ovelhas Ideal durante o Anestro Estacional

The objective was to determine the plasma levels of the triiodothyronine (T-3) and thyroxine (T-4), in six Ideal ewe sheeps during seasonal anoestrus, by radioimmunoassay. The blood samples were collected from the jugular vein catheter over a period of 24 hours, at 2-h intervals (12, 14, 16, IX, 20, 22, 24, OZ, 04, 06, OX, and 10 hours). II was used analysis of variance for data adjustments, and the decomposition of the collected periods in non-orthogonal polynomial. It was observed a crescent linear effect of the T-4 plasma levels, presenting lower level of 49.60+/-11.41 and higher level of 76.99+/-10.0 ng/ml, at fourth and last blood samples collection, respectively. T-3 plasma levels showed a significant cubic response during 24 hours, with the highest value of 0.94+/-0.20 ng/ml at the end of the the study. In conclusion, thyroidal hormones levels were lower in the first blood samples collection, increasing progressively during the night, reaching the highest levels at the morning period.

Evaluation testicular fine needle aspiration cytology and serum testosterone levels in dogs

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

O aumento em superfície em Adelia membranifolia (Müll. Arg.) Pax & K. Hoffm. e Peltophorum dubium (Spreng.) Taub., em resposta ao estresse por deficiência nutricional e alagamento do substrato

Foram estudadas as respostas plásticas em plântulas e plantas jovens de duas espécies arbóreas nativas, Adelia membranifolia (Müll. Arg.) Pax & K. Hoffm. (Euphorbiaceae) e Peltophorum dubium (Spreng.) Taub. (Leguminosae-Caesalpinoidae), submetidas ao alagamento do substrado associado ao déficit nutricional. Avaliou-se o acréscimo em superfície no módulo de expressão, definido como a região do eixo da plântula que, sob condições de estresse, sofre rediferenciação sendo considerada a hipertrofia lenticelar como a expressão morfogenética mais comum. em A. membranifolia a associação do estresse nutricional e alagamento do substrato foi a condição mais restritiva tanto para o crescimento em altura e diâmetro do colo quanto para as expressões morfogenéticas. Já em P. dubium o estado nutricional foi mais restritivo que o alagamento para o crescimento do colo. em geral, para ambas as espécies, plantas nutridas mantidas alagadas restabeleceram seu ritmo de crescimento a partir do terceiro mês de tratamento, além de expressarem respostas plásticas como a hipertrofia lenticelar e a formação de raízes adventícias. Quanto ao aumento em superfície, nos lotes desnutridos, verificou-se que em A. membranifolia de um total de 38% de superfície expressa, 12% corresponderam à hipertrofia lenticelar. em P. dubium verificou-se as maiores variações de aumento em superfície expressa por hipertrofia lenticelar, elevando de 5% para 13%. em geral a superfície modular nos lotes tratados foi o dobro em relação ao controle. É possível que o aumento em superfície, para algumas espécies, seja significativo como tendência ao aumento da estabilidade no desenvolvimento a partir de interações fisiológicas.

Foraging Behavior of the Swarm-founding Wasp Polybia (Trichothorax) sericea (Hymenoptera, Vespidae): Daily Resource Collection Activity and Flight Capacity

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)