Expressão de zif-268 durante a aquisição, evocação e extinção de uma memória aversiva

In the behavioral paradigm of discriminative avoidance task, both short and long-term memories have been extensively investigated with behavioral and pharmacological approaches. The aim of the present study was to evaluate, using the abovementioned model, the hippocampal expression of zif-268 - a calcium-dependent immediate early gene involved with synaptic plasticity process - throughout several steps of memory formation, such as acquisition, evocation and extiction. The behavioral apparatus consisted of a modified elevaated plus-maze, with their enclosed arms disposed in "L". A pre-exposure to the maze was made with the animal using all arms enclosed, for 30 minutes, followed by training and test, during 10 minutes each. The between sections interval was 24h. During training, aversive stimuli (bright light and loud noise) were actived whenever the animals entered one of the enclosed armas (aversive arm). Memory acquisiton, retention and extinction were evaluated by the percentage of the total time spent exploring the aversive arm. The parameters evaluated (time spent in the arms and total distance traveled) were estimated with an animal tracking software (Anymaze, Stoelting, USA). Learning during training was estimated by the decrease of the time spent exploring the aversive arm. One hour after the beginning of each section, animals were anaesthetized with sodium-thiopental (i.p.) and perfused with 0.9% heparinized saline solution followed by 4% paraformaldehyde. Brains were cryoprotected with 20% sucrose, separeted in three blocks and frozen. The middle block, containing the hippocampus, was sectioned at 20 micro meters in the coronal plane and the resutant sections were submitted to zif-268 immunohistochemistry. Our results show an increased expression of zif-268 in the dentate gyrus (DG) during the evocation and extinction stages. There is a distinct participation of the DG during the memory evocation, but not during its acquisition. Inaddition, all hippocampal regions (CA1, CA3 and DG) presented an increased zif-268 expression during the process of extinction.

Antidepressivos modificam a extinção de uma memória aversiva em ratas

Treatment of major depression, posttraumatic stress disorder and other psychopathologies with antidepressants can be associated with improvement of the cognitive deficits related to these disorders. Although the mechanisms of these effects are not completely elucidated, alterations in extinction of aversive memories are believed to be present in these psychopathologies. Moreover, researches with laboratory animals usually focus on male subjects, and we have recently verified that extinction of an aversive task is reduced in female rats when compared to males. In the present study, female rats were long-term treated with clinically used antidepressants (fluoxetine, nortriptyline or mirtazapine) and tested in the plus-maze discriminative avoidance and forced swimming tests in order to evaluate learning, memory, extinction, anxiety and depression-related behaviors. All groups learned the task, but learning was somewhat faster in nortriptyline and mirtazapine-treated animals . Task retrieval was also showed by all experimental groups. Chronic treatment with fluoxetine, but not with the other antidepressants, increased extinction of the discriminative task. In the forced swimming test, animals treated with fluoxetine and mirtazapine showed decreased immobility duration. In conclusion, antidepressants interfere with learning and female rats treated with fluoxetine presented increased extinction of the aversive memory task. On the other hand, both fluoxetine and mirtazapine were effective in the forced swimming test, suggesting dissociation between the antidepressant effects and the extinction of aversive memories

Influência do ciclo estral no efeito do diazepam na ansiedade e memória de ratas

Memory and anxiety are related phenomena. Several evidences suggest that anxiety is fundamental for learnining and may facilitate or impair the memory formation process depending of the context. The majority of animal studies of anxiety and fear use only males as experimental subjects, while studies with females are rare in the literature. However, the prevalence in phobic and anxiety disorders is greater in women than in men. Moreover, it is known that gender maybe influence benzodiazepine effects, the classic drugs used for anxiety disorders treatment. In this respect, to further investigate if fear/anxiety aspects related to learning in female subjects would contribute to the study of phobic and anxiety disorders and their relationship with learning/memory processes, the present work investigates (a) the effects of benzodiazepine diazepam on female rats performance in a aversive memory task that assess concomitantly anxiety/emotionality, as the interaction between both; (b) the influence of estrous cycle phases of female rats on diazepam effects at aversive memory and anxiety/emotionality, and the interaction between both and (c) the role of hormonal fluctuations during estrous cycle phases in absence of diazepam effects in proestrus, because female rats in this phase received or not mifepristone, the antagonist of progesterone receptor, previously to the diazepam treatment. For this purpose, the plus maze discriminative avoidance task, previously validated for studies of anxiety concomitantly to learning/memory, was used. The apparatus employed is an adaptation of a conventional plus maze, with two opens arms and two closed arms, one of which presenting aversive stimulation (noise and light). The parameters used were: time in non-aversive arm compared to time in aversive and percentage of time in aversive arm on several temporal divisions, in order to evaluate memory; percentage of time in open arms, risk assessment, head dipping and end exploring to evaluate anxiety ; and distance traveled for locomotion. In experiment I, we found anxiolytic effect of diazepam only for 4 mg/kg dose, however the amnestic effect appear at a dose of 2 mg/kg. In second experiment, rats were divided in groups according estrous cycle phase (metaestrus/diestrus, proestrus e estrus). In this experiment, when we considered estrous cycle phase or diazepam treatment, the results did not demonstrate any differences in anxiety/emotionality parameters. The amnestic effects of diazepam occur in female rats in metestrus/diestrus and estrus and is absent in proestrous rats. Proestrous female rats that received mifepristone exhibited the amnestic effect of diazepam and also anxiolytic effects, that it was not previously observed in this dose. The results have demonstrated dissociation of anxiolytic and amnestic diazepam effects, not previously observed in males; the absence of amnestic effect of diazepam in proestrous phase; and the possible role of progesterone in aversive memory over diazepam effect, because the mifepristone, associated with diazepam, caused amnestic effect in proestrus

Alterações na evocação de uma memória aversiva ao liongo do ciclo estral de ratas: influências da transmissão gabaérgica na amígdala

GABAergic neurotransmission has been implicated in many aspects of learning and memory, as well as mood and anxiety disorders. The amygdala has been one of the major focuses in this area, given its essential role in modulating emotionally relevant memories. However, studies with male subjects are still predominant in the field. Here we investigated the consequences for an aversive memory of enhancing or decreasing GABAergic transmission in the basolateral nucleus of the amygdala (BLA). Wistar female rats were trained in the plus-maze discriminative avoidance task, in which they had to learn to avoid one of the enclosed arms where an aversive stimulus consisting of a bright light and a loud noise was given (day 1). Fifteen minutes before the test session (day 2) animals received 0,2 μL infusions of either saline solution, the GABAergic agonist muscimol (0,05 mg/ml), or the GABAergic antagonist bicuculine (0,025 mg/ml) bilaterally intra-BLA. On the test day, females in proestrous or estrous presented adequate retrieval and did not extinguish the task, while females in metestrous or diestrous presented impaired retrieval. In the first group, muscimol infusion impaired retrieval and bicuculline had no effect, suggesting naturally low levels of GABAergic transmission in the BLA of proestrous and estrous females. In the second group, muscimol infusion had no effect and bicuculline reversed retrieval impairment, suggesting naturally high levels of GABAergic transmission in the BLA of metestrous and diestous females. Additionally, proestrous and estrous females presented higher anxiety levels compared to metestrous and diestrous females, which could explain better performance of this group. On the other hand, BLA GABAergic system did not interfere with the innate fear response because drug infusions had no effect in anxiety. Thus, retrieval alterations caused by the GABAergic drugs were probably related specifically to memory processes

Álcool e comportamento: efeitos na aprendizagem e memória

Excessive alcohol consumption is responsible for many harmful effects on individuals and society. Despite years of research, the mechanisms by which alcohol affects neurological functions and the exact causes of cognitive impairment related to long-term use are unknown. In this sense, this master study proposed to observe how different doses of alcohol affect the addiction response and the learning ability of two fish species: Betta splendens and Danio rerio, the latter a commonly model due to organizational and functional characteristics shared with mammals. For this, different concentrations of ethanol (0%, 0.1%, 0.25%, 1% and 1.5%) were used in acute, chronic and withdrawal treatments. We tested the fish in three experimental protocols: 1) alcohol addiction potential using conditioned place preference, 2) associative conditioning using light as unconditioned stimulus and food as conditioned stimulus and 3) spatial learning using a maze without cues. For the alcohol addiction potential, preference between two different places in a shuttle box was tested before and after alcohol exposure (chronic and acute). In this test, the animals intoxicated by 0.1% did not change behavior, while animals receiving 1% and 1.5% alcohol changed the initial preference to the side where they received alcohol For the associative conditioning, the results show that the groups undergoing low dose (0.1%), both in chronic and withdrawal treatment, learned the task faster than control; groups under 0.25 and 1% alcohol withdrawal learned the task after control; groups chronically intoxicated with these doses did not learn the task. For the spatial learning test, fish submitted to acute and chronic treatments decreased the time to exit the maze; there were significant differences in the animal s performance in a dose-dependent pattern. This difference was not observed for the withdrawal treatment. Given these results, we conclude that the effects of alcohol on learning are dependent on the dosage. Furthermore, low doses of alcohol seem to maximize animal performance on learning tasks and do not alter their seeking behavior, while higher doses induced addition and hinder learning

Aprendizagem espaço-temporal e efeitos das condições luminosas no aprendizado do peixe paulistinha

Para processar a informação ambiental e perceber o tempo, os indivíduos utilizam-se de pistas ambientais, como luz e temperatura, que servem como guias para o relógio interno. O mecanismo temporizador endógeno é chamado relógio circadiano, o qual comanda uma grande variedade de ritmos diários bioquímicos, fisiológicos e comportamentais presentes nos organismos. Com isso, os animais podem antecipar eventos espaço-temporalmente distribuídos e usar essa informação para organizar as atividades diárias, o que é uma vantagem adaptativa para os indivíduos, já que muitos fatores ambientais apresentam variação circadiana. Aprendizagem espaço-temporal (do inglês: "time-place learning’’-TPL) é a habilidade de associar lugares com importantes eventos biológicos em diferentes horas do dia. Em nosso estudo utilizamos como modelo o peixe paulistinha (Danio rerio), conhecido por ser altamente social, para testar aprendizagem espaço-temporal baseada em reforço social. Além disso, objetivamos averiguar os efeitos das condições de claro constante e escuro constante na aprendizagem espaço-temporal, e se nessas condições, a atividade do peixe paulistinha é alterada. Para isso, testamos três diferentes condições (n=10): grupo claro-escuro (CE), grupo claro constante (CC) e grupo escuro constante (EE) durante 30 dias da seguinte maneira: diariamente, um grupo de 5 peixes paulistinha foi introduzido em um recipiente localizado no compartimento da manhã (um dos lados do aquário), às 8:00h e retirado às 9:00h, e em outro recipiente do compartimento da tarde (lado oposto do aquário), às 17:00h e removido às 18:00h, servindo como estímulo para que o peixe experimental ocupasse o compartimento onde o grupo fosse colocado. O comportamento foi filmado nos dois horários, 15 minutos antes e durante os 60 minutos de exposição ao estímulo, no 15º e no 30ª dia, porém neste último, os peixes foram filmados sem a presença do estímulo a fim de averiguarmos a aprendizagem espaço-temporal. Por fim, para saber a influência das três condições luminosas na atividade dos peixes, filmamos os últimos 6 dias de teste, para registrar o padrão de atividade. Nossos resultados mostraram que em ciclo claro-escuro (CE) o peixe paulistinha apresenta TPL, bem como é capaz de antecipar a hora e local do estímulo (grupo de coespecíficos), enfatizando a importância do estímulo social para a aprendizagem. Em condições de claro constante e escuro constante, o peixe paulistinha não apresentou aprendizagem espaço-temporal. Ademais, após 30 dias em condições luminosas constantes (claro constante e escuro constante), o peixe paulistinha mantém ritmo circadiano, porém em claro constante sua atividade é aumentada e seu ritmo atividade-repouso é alterado, através de um padrão de atividade distribuída homogeneamente ao longo das 24h, ao invés de concentrada na subjetiva fase clara, como nos grupos de ciclo claro-escuro e escuro constante, os quais conservam o padrão de atividade diurno da espécie.

Identidade, memória e narrativas na dança de São Gonçalo do povoado Mussuca (SE)

This study deals with the participation of the dance of São Gonçalo of the Mussuca town/SE, in the process of construction of the ethnic identity among this social group. The Mussuca is a grouping recognized as afro-descendents, linked with black enslaved people in the valley of the Cotinguiba region. The collective memory functions as a drive of this linking with the past and if it makes to elaborate narratives on this descent. The objective of this study was to investigate the ways the rite went through to constitute itself as an element of ethnic representation. Internal and external agents had been identified who had participated in different contexts. By means of an ethnographic work we ve reached some aspects of the local structure social which demonstrated the contradictions through the social relations of the group. This process of ethnic autorecognition presents the kinship and the space question as definers of the social arrangements which establish its ethnic boundaries