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BACKGROUND: In studies that measure social behavior of a freely interacting pair rats social behavior of one rat is strongly influenced by the behavior of the other. This prevents evaluating social behavior of one single rat. NEW METHOD: We assessed the motivation to interact socially in a modified open-field, by measuring the time a rat attempted to interact with a co-specific separated by a grid in a birdcage outside of the apparatus. We propose time in front of the birdcage is an indicator of social behavior. RESULTS: We showed that the focal rat allocates more time in front of the birdcage, interacting with another rat through the grid. Also, that the presence of the other rat that attracts the focal rat. Habituation to the apparatus, repeated testing and illumination condition did not alter the proximity measures of rats. Finally, treatment with chlordiazepoxide (3.0mg/kg) either increased the time spent in front of the cage by males and females or (5.6mg/kg) increased the proximity measure of females. COMPARING WITH EXISTING METHOD: Our method prevents partners from influencing the target rat's social behavior; existing methods do not. Also, it is more sensitive to the effect of chlordiazepoxide than the broadly used method proposed by File and Hyde (1978). CONCLUSIONS: Proximity is an advantageous measure: it allows the assessment of only one focal animal without the interference of a partner; it is simple to take; it requires little interpretation skills or training from the experimenter, no special equipment or conditions.
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Conducta Animal , Modelos Psicológicos , Ratas Wistar/psicología , Conducta Social , Conducta Espacial , Animales , Conducta Animal/efectos de los fármacos , Clordiazepóxido/farmacología , Femenino , Habituación Psicofisiológica , Iluminación , Masculino , Motivación/efectos de los fármacos , Pruebas Psicológicas , Psicotrópicos/farmacología , Distribución Aleatoria , Reproducibilidad de los Resultados , Conducta Espacial/efectos de los fármacos , Factores de TiempoRESUMEN
ABSTRACT The present technical note introduces the X-PloRat, a software for the scoring of animal location, displacement, and other behavioral properties, in enclosed spaces. The X-PloRat offers a variety of data reports and can be used for training students in behavioral observation and in validating other kinds of scoring procedures.
RESUMO A presente nota técnica apresenta o X-PloRat, um software que permite o registro da localização, deslocamento e outras propriedades do comportamento animal em espaços confinados. O X-PloRat oferece uma grande variedade de relatórios e pode ajudar no treinamento de estudantes em observação do comportamento e também na validação de outros tipos de registros.
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The elevated plus-maze is an apparatus widely used to study the level of anxiety in rodents. The maze is plus-shaped, with two enclosed arms and two open arms, and elevated 50cm from the floor. During a test, which usually lasts for 5min, the animal is initially put at the center and is free to move and explore the entire maze. The level of anxiety is measured by variables such as the percentage of time spent and the number of entries in the enclosed arms. High percentage of time spent at and number of entries in the enclosed arms indicate anxiety. Here we propose a computational model of rat behavior in the elevated plus-maze based on an artificial neural network trained by a genetic algorithm. The fitness function of the genetic algorithm is composed of reward (positive) and punishment (negative) terms, which are incremented as the computational agent (virtual rat) moves in the maze. The punishment term is modulated by a parameter that simulates the effects of different drugs. Unlike other computational models, the virtual rat is built independently of prior known experimental data. The exploratory behaviors generated by the model for different simulated pharmacological conditions are in good agreement with data from real rats.
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Ansiedad/fisiopatología , Simulación por Computador , Conducta Exploratoria/fisiología , Aprendizaje por Laberinto/fisiología , Redes Neurales de la Computación , Algoritmos , Animales , Ansiolíticos/farmacología , Ansiedad/inducido químicamente , Ansiedad/tratamiento farmacológico , Estimulantes del Sistema Nervioso Central/farmacología , Conducta Exploratoria/efectos de los fármacos , Aprendizaje por Laberinto/efectos de los fármacos , Pruebas Neuropsicológicas , Ratas , Recompensa , Índice de Severidad de la EnfermedadRESUMEN
A behavioral test battery is proposed for wall lizards (Tropidurus oreadicus) that consists of inducing tonic immobility (TI) followed by post-TI behavioral scoring. After the induction of TI, the usual behavioral sequence was flight followed by freezing and tongue-flicking and/or thigmotaxis, with flight being more probable than freezing. These sequences were not observed after restraint in a normal upward position (which induced freezing but not TI) or after handling (which increased the probability of tongue-flicking). Alprazolam and imipramine selectively decreased the duration of TI as well as the following flight and freezing behavior. Tongue-flicking was increased by diazepam and alprazolam, whereas fluoxetine decreased it. Finally, thigmotaxis was reduced by diazepam, alprazolam, and imipramine but increased by fluoxetine. These results suggest that panic and anxiety can be discriminated pharmacologically in wall lizards...
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Animales , Pérdida de Tono Postural , Ansiedad , Benzodiazepinas , Lagartos , PánicoRESUMEN
The elevated plus maze is a widely used experimental test to study anxiety-like rodent behavior. It is made of four arms, two open and two closed, connected at a central area forming a plus shaped maze. The whole apparatus is elevated 50 cm from the floor. The anxiety of the animal is usually assessed by the number of entries and duration of stay in each arm type during a 5-min period. Different mathematical methods have been proposed to model the mechanisms that control the animal behavior in the maze, such as factor analysis, statistical inference on Markov chains and computational modeling. In this review we discuss these methods and propose possible extensions of them as a direction for future research.
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Conducta Animal/fisiología , Matemática , Aprendizaje por Laberinto , Modelos Animales , Animales , Ansiedad , RoedoresRESUMEN
The so-called anxiolytic and anxiogenic drugs are considered to cause, respectively, increases and decreases in plus-maze open arm exploration, without modifying locomotor activity occurring in the closed arms in an elevated plus-maze when the animals are tested in an illuminated environment. Simply testing animals in the dark also increases open arm exploration, which may be interpreted as an anxiolytic effect. We investigated the effects of two GABAergic drugs, pentylenetetrazol (10 and 20 mg/kg) and chlordiazepoxide (1.5 and 3 mg/kg), and one non-GABAergic drug, caffeine (10 and 30 mg/kg) on anxiety levels of rats tested in the elevated plus-maze under two illumination conditions, light or dark. All animals explored more the open arms in the dark. In the light, pentylenetetrazol decreased open arm exploration while chlordiazepoxide had the opposite effect. Neither pentylenetetrazol nor chlordiazepoxide had any effect in the dark. Caffeine, increased open arms exploration in both illumination conditions. These results indicate that light triggers aversion, a response mediated by GABA since the GABAergic drugs, but not caffeine, were ineffective when the rats were tested in the dark.
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Cafeína/farmacología , Clordiazepóxido/farmacología , Iluminación/métodos , Aprendizaje por Laberinto/efectos de los fármacos , Pentilenotetrazol/farmacología , Animales , Relación Dosis-Respuesta a Droga , Conducta Exploratoria/efectos de los fármacos , Masculino , Ratas , Ratas WistarRESUMEN
The elevated plus-maze is an animal model of anxiety used to study the effect of different drugs on the behavior of the animal. It consists of a plus-shaped maze with two open and two closed arms elevated 50cm from the floor. The standard measures used to characterize exploratory behavior in the elevated plus-maze are the time spent and the number of entries in the open arms. In this work, we use Markov chains to characterize the exploratory behavior of the rat in the elevated plus-maze under three different conditions: normal and under the effects of anxiogenic and anxiolytic drugs. The spatial structure of the elevated plus-maze is divided into squares, which are associated with states of a Markov chain. By counting the frequencies of transitions between states during 5-min sessions in the elevated plus-maze, we constructed stochastic matrices for the three conditions studied. The stochastic matrices show specific patterns, which correspond to the observed behaviors of the rat under the three different conditions. For the control group, the stochastic matrix shows a clear preference for places in the closed arms. This preference is enhanced for the anxiogenic group. For the anxiolytic group, the stochastic matrix shows a pattern similar to a random walk. Our results suggest that Markov chains can be used together with the standard measures to characterize the rat behavior in the elevated plus-maze.
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Conducta Exploratoria/fisiología , Cadenas de Markov , Aprendizaje por Laberinto/fisiología , Animales , Ansiolíticos/farmacología , Clordiazepóxido/farmacología , Convulsivantes/farmacología , Conducta Exploratoria/efectos de los fármacos , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Pentilenotetrazol/farmacología , Ratas , Ratas WistarRESUMEN
Although several reports have demonstrated physiological and behavioral changes in adult rats due to neonatal immune challenges, little is known about their effects in adolescence. Since neonatal exposure to lipopolysaccharide (LPS) alters the neural substrates involved in cognitive disorders, we tested the hypothesis that it may also alter the response to novel environments in adolescent rats. At 3 and 5 days of age, male Wistar rats received intraperitoneal injections of either vehicle solution or E. coli LPS (0.05mg/kg) or were left undisturbed. In the mid-adolescent period, between 40 and 46 days of age, the rats were exposed to the following behavioral tests: elevated plus-maze, open-field, novel-object exploration task, hole-board and the modified Porsolt forced swim test. The results showed that, in comparison with control animals, LPS-treated rats exhibited (1) less anxiety-related behaviors and enhanced patterns of locomotion and rearing in the plus-maze and the open-field tests, (2) high levels of exploration of both objects in the novel-object task and of corner and central holes in hole-board test, and (3) more time spent diving, an active behavior in the forced swim test. The present findings suggest that neonatal LPS exposure has long-lasting effects on the behavior profile adolescent rats exhibit in response to novelty. This behavioral pattern, characterized by heightened exploratory activity in novel environments, also suggests that early immune stimulation may contribute to the development of impulsive behavior in adolescent rats.
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Ansiedad , Conducta Exploratoria/efectos de los fármacos , Lipopolisacáridos/farmacología , Actividad Motora/efectos de los fármacos , Análisis de Varianza , Animales , Conducta Animal/efectos de los fármacos , Masculino , Ratas , Ratas Wistar , NataciónRESUMEN
Zebrafish are increasingly being used in behavioral neuroscience, neuropsychopharmacology and neurotoxicology. Recently, behavioral screens used to model anxiety in rodents were adapted to this species, and novel models which tap on zebrafish behavioral ecology have emerged. However, model building is an arduous task in experimental psychopathology, and a continuous effort to assess the validity of these measurements is being chased among some researchers. To consider a model as valid, it must possess face, predictive and/or construct validity. In this article, we first review some notions of validity, arguing that, at its limit, face and predictive validity reduce to construct validity. Then we review some procedures which have been used to study anxiety, fear or related processes in zebrafish, using the validity framework. We conclude that, although the predictive validity of some of these models is increasingly being met, there is still a long way in reaching the desired level of construct validity. The refinement of models is an ongoing activity, and behavioral validation and parametric research ought to advance that objective.
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Ansiedad/psicología , Modelos Animales , Reproducibilidad de los Resultados , Pez Cebra , Animales , HumanosRESUMEN
The scototaxis (dark/light preference) protocol is a behavioral model for fish that is being validated to assess the antianxiety effects of pharmacological agents and the behavioral effects of toxic substances, and to investigate the (epi)genetic bases of anxiety-related behavior. Briefly, a fish is placed in a central compartment of a half-black, half-white tank; following habituation, the fish is allowed to explore the tank for 15 min; the number and duration of entries in each compartment (white or black) are recorded by the observer for the whole session. Zebrafish, goldfish, guppies and tilapias (all species that are important in behavioral neurosciences and neuroethology) have been shown to demonstrate a marked preference for the dark compartment. An increase in white compartment activity (duration and/or entries) should reflect antianxiety behavior, whereas an increase in dark compartment activity should reflect anxiety-promoting behavior. When individual animals are exposed to the apparatus on only one occasion, results can be obtained in 20 min per fish.
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Ansiedad/fisiopatología , Conducta Animal/fisiología , Conducta Exploratoria , Peces/fisiología , Habituación Psicofisiológica , Animales , Conducta Animal/efectos de los fármacos , Clordiazepóxido/farmacología , Oscuridad , Conducta Exploratoria/efectos de los fármacos , Carpa Dorada/fisiología , Luz , Poecilia/fisiología , Valor Predictivo de las Pruebas , Psicometría/métodos , Tilapia/fisiología , Pez Cebra/fisiologíaRESUMEN
Scototaxis, the preference for dark environments in detriment of bright ones, is an index of anxiety in zebrafish. In this work, we analyzed avoidance of the white compartment by analysis of the spatiotemporal pattern of exploratory behavior (time spent in the white compartment of the apparatus and shuttle frequency between compartments) and swimming ethogram (thigmotaxis, freezing and burst swimming in the white compartment) in four experiments. In Experiment 1, we demonstrate that spatiotemporal measures of white avoidance and locomotion do not habituate during a single 15-min session. In Experiments 2 and 3, we demonstrate that locomotor activity habituates to repeated exposures to the apparatus, regardless of whether inter-trial interval is 15-min or 24-h; however, no habituation of white avoidance was observed in either experiment. In Experiment 4, we confined animals for three 15-min sessions in the white compartment prior to recording spatiotemporal and ethogram measures in a standard preference test. After these forced exposures, white avoidance and locomotor activity showed no differences in relation to non-confined animals, but burst swimming, thigmotaxis and freezing in the white compartment were all decreased. These results suggest that neither avoidance of the white compartment nor approach to the black compartment account for the behavior of zebrafish in the scototaxis test.
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Ansiedad , Actividad Motora , Animales , Ambiente , Conducta Exploratoria , Femenino , Reacción Cataléptica de Congelación , Habituación Psicofisiológica , Iluminación , Masculino , Natación , Factores de Tiempo , Pez CebraRESUMEN
The effect of intraseptal injections of lidocaine before a first or a second session in the elevated plus-maze, in a test-retest paradigm, was investigated. In addition to gross session analyses, a minute-by-minute analysis of the sessions was used to evaluate both anxiety and memory. Lidocaine injections before the test session produced increases in the frequency of entries, time spent and distance run in the open arms without affecting activity occurring in the closed arms. During the retest session, saline- and lidocaine-treated rats exhibited increased indices of anxiety and lidocaine-treated rats exhibited decreased closed-arm entries. The minute-by-minute analysis showed a faster decrease in anxiety-related behaviors during the test session by saline- than by lidocaine-treated rats and a significant decrease in closed-arm exploration by saline-treated rats, but not by lidocaine-treated ones. Lidocaine injection before the retest session produced increases in the frequency of entries, time spent and distance run in the open arms in the second session when compared with saline-treated rats. Minute-by-minute analysis showed an increase in the time spent in the open arms by lidocaine animals at the beginning of the retest session in comparison to saline animals and a significant decrease in closed-arm exploration by both groups. These results suggest that inactivation of the medial septum by lidocaine affects the expression of unconditioned and conditioned forms of anxiety in the elevated plus-maze and, in a lesser way, the acquisition and retention of spatial information.
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Conducta Exploratoria/fisiología , Aprendizaje por Laberinto/fisiología , Tabique del Cerebro/fisiología , Anestésicos Locales/administración & dosificación , Anestésicos Locales/farmacología , Animales , Ansiedad/fisiopatología , Lidocaína/administración & dosificación , Lidocaína/farmacología , Masculino , Memoria/fisiología , Pruebas Neuropsicológicas , Ratas , Ratas Wistar , Tabique del Cerebro/efectos de los fármacos , Factores de TiempoRESUMEN
In order to determine the modulation of anxiolytic and panicolytic-like effects of diazepam by the hormonal cycle of female rats, male and female rats - the latter divided per estrous cycle phase (estrus, diestrus, metaestrus and proestrus) - were tested in the elevated T-maze, a behavioral model of panic and anxiety. Diazepam (0.5, 1.0 and 2.0 mg/kg) or saline solution was injected in individual animals that were submitted to one session in the elevated T-maze 25 min after drug/saline administration. The test consisted of three avoidance trials and one escape trial, separated by a 30 s interval, during which the animals were isolated in individual cages. The avoidance trials began with the animal being placed at the end of the maze's enclosed arm. The time necessary for the animal to leave the central square was considered as the response's latency. The trials that exceeded 300 s were considered as failures. Results demonstrate a decrease in the effects of diazepam in inhibitory avoidance (anxiety) trials in females in diestrus and proestrus, but no relation of gender or estrous cycle on diazepam effects on escape trials (fear). The results support the hypothesis that down-regulation of GABA A receptors by activation of nuclear estrogen receptors and induction of PKC-mediated GABA A receptor phosphorylation by activation of surface estrogen receptors in raphe neurons underlie the modulation of diazepam sensitivity by estrogen.
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Animales , Ratas , Ansiedad , Ciclo Estral , Diazepam/farmacología , EstrógenosRESUMEN
The elevated plus-maze is a device widely used to assess rodent anxiety under the effect of several treatments, including pharmacological agents. The animal is placed at the center of the apparatus, which consists of two open arms and two arms enclosed by walls, and the number of entries and duration of stay in each arm are measured for a 5-min exposure period. The effect of an anxiolytic drug is to increase the percentage of time spent and number of entries into the open arms. In this work, we propose a new measure of anxiety levels in the rat submitted to the elevated plus-maze. We represented the spatial structure of the elevated plus-maze in terms of a directed graph and studied the statistics of the rat's transitions between the nodes of the graph. By counting the number of times each transition is made and ordering them in descending frequency we represented the rat's behavior in a rank-frequency plot. Our results suggest that the curves obtained under different pharmacological conditions can be well fitted by a power law with an exponent sensitive to both the drug type and the dose used.
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Ansiolíticos/farmacología , Conducta Animal/fisiología , Etología/métodos , Aprendizaje por Laberinto/fisiología , Neurofarmacología/métodos , Neuropsicología/métodos , Animales , Ansiedad/tratamiento farmacológico , Ansiedad/psicología , Conducta Animal/efectos de los fármacos , Interpretación Estadística de Datos , Etología/instrumentación , Conducta Exploratoria/efectos de los fármacos , Conducta Exploratoria/fisiología , Vivienda para Animales , Matemática , Aprendizaje por Laberinto/efectos de los fármacos , Modelos Animales , Modelos Teóricos , Neurofarmacología/instrumentación , Neuropsicología/instrumentación , RatasRESUMEN
The positive reinforcing properties of addictive drugs have a primary role in the development of drug dependence. In the field of opiates, great attention has been given to this phenomenon, as well as to the negative properties eliciting craving and inducing relapse during withdrawal. This study was designed to evaluate whether elevated plus-maze (EPM) experienced rats withdrawn from low doses of subcutaneous (s.c.) morphine (10 mg/kg), in which a high anxiety level is the most prominent withdrawal symptom, acquire place preference when submitted to a conflict paradigm in which drug effects are paired with an aversive context: the distal part of the open arms of an EPM. Both the anxiety test and place preference conditioning were measured in the same apparatus, a biased version of the plus-maze. In order to verify the influence of previous EPM spatial learning on the performance of morphine-withdrawn rats, half the animals in this study experienced the EPM prior to treatment. Additional groups were also tested under the influence of morphine effects. The effects of the treatments were quantified through the analysis of three types of measures: anxiety was inferred from the use of the conventional measures (percentage of entries and time spent in the open-arms) and risk-assessment behaviours (frequency of stretched-attending postures - SAP, and time spent at the centre of the maze). Place preference conditioning was evaluated through analysis of the number of entries, total time spent and distance run in the open-arm extremities, which is where the animals were conditioned. The number of closed-arm entries was taken as an index of locomotor activity. Our results showed that (i) EPM naïve rats pre-treated with morphine did not develop place preference, behaving like control rats; (ii) rats that had previous experience in the EPM showed no changes in open-arm avoidance on the second exposure, when compared with rats naïve for this condition; (iii) previous spatial learning of the EPM contextual cues was, in fact, a requirement for anxiety-inducing place preference for the open-arms in morphine-withdrawn rats and, (iiii) conditioned place preference was achieved both in rats under the effects of morphine and in withdrawal, probably through the influence of the positive or negative reinforcing effects promoted by the presence or absence of the drug in the central nervous system.
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Aprendizaje por Laberinto , Morfina , Síndrome de Abstinencia a SustanciasRESUMEN
The present work aimed at studying the influence of the estrous cycle in the forced swim test, an animal model of depression. For this, 44 male and female Wistar rats were divided into five groups according to the hormonal state in the first day of the study: metaestrus (N = 12), diestrus (N = 8), proestrus (N = 7), estrous (N = 6) and males (N = 11). They were housed in groups of five, with water and food ad libitum under a 12/12 h light/dark cycle. Females were screened daily for the estrous cycle. The animals were subjected to two swimming sessions in a glass cylinder with water up to 15 cm at 28±2º C. The data of the first five minutes of a 15-min first session were compared to those of a 5-min second session 24 h later. The results indicate that the latency to the first immobility was substantially reduced in the second session and was longer for females in diestrus and proestrus in the first session. The results also indicate that females in diestrus and proestrus exhibited less immobility than males in the first session; females in diestrus also exhibited less immobility than females in metaestrus. Females in metaestrus and diestrus, as well as males, did not present the decrease in total immobility times in the second session. The present results are analyzed in terms of differential effects of progesterone and estrogen on a learning component and an affective component.
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The are few reports on the relationship between elevated plus-maze and effects of maintenance conditions in the days prior to the test. Previously, we have demonstrated that being forcibly in total dark or in light for four days does not alter exploratory behavior in the plus-maze. The present study aimed at recording illumination level preferences in rats using a box with light and dark compartments (or another with two light compartments) and the behavioral effect of this choice on the plus-maze. The rats allowed to express their preference to remain in one particular level of illumination tended to stay initially in the dark compartment and gradually preferred to remain in the light compartment. When tested in the elevated plus-maze there were no effects in comparison to controls kept in commercial cages.
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The set of answers emitted by the organisms in response to a real or potential danger is called the Anxiety State. This state, of evolutionary importance, appears when a strong danger or risk is detected, and in response to ambiguous threat stimuli that could be of innate or learned nature. A low level of anxiety is beneficial for the animals as a fundamental resource of protection of the individual against physical and social dangers. The neural basis responsible by such states would be the oldest cerebral areas, preserved in many species, that are crucial for the control of the emotions, and whosemalfunctions lead to mood disorders. Many experimental models have been developed to help the study of behaviors and neural basis of the anxiety state, providing tools that collaborate not only to the therapeutics of pathologies, but also to the better understanding of the world of the emotions.
El conjunto de respuestas emitidas por diferentes especies frente a un peligro real o potencial se conoce como Estado de Ansiedad. Este estado, de importancia evolutiva para las especies, aparece cuando se detecta un peligro o amenaza o ante la presencia de estímulos ambiguos, innatos o aprendidos, que indican una amenaza. Así, un determinado nivel de ansiedad es benéfico para los animales, siendo un recurso fundamental de protección contra peligros físicos y sociales. Los substratos neurales responsables por tales estados, corresponden a áreas cerebrales antiguas, preservadas en muchas especies y críticas para el control de las emociones que al sufrir alteraciones en su funcionamiento producen disturbios del comportamiento. Diversos modelos experimentales se han desarrollado para estudiar los componentes conductuales y sustratos neurales implicados en el estado de ansiedad y su utilización ya está suministrando herramientas para complementar la comprensión de diversas patologías y conocimiento del mundo de las emociones.
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Prenatal stress is a putative model for studying some psychopathological disorders. Indeed, submitting pregnant animals to stress leads to enhanced anxiety in the adult offspring. However, little is known about how prenatal stress effects interacts with anxiety throughout development. To study this issue, prenatally stressed rats were tested in the elevated plus-maze at different ages. During pregnancy female rats were submitted to uncontrollable electric foot shock sessions every other day or kept undisturbed (controls). After delivery, litters from control and stressed dams were left undisturbed from the 3rd to the 14th postnatal days. Male and female rats were tested in the elevated plus-maze at the ages of 30, 45 or 60 days. The following measures were taken in the elevated plus-maze: number of entries and time spent in the arms (or their extremities) and frequency and time spent in naturalistic behaviors (stretching, rearing, end exploring, grooming and head dipping). Decreases in the percentage of entries into and in the time spent (only females) in the open arms were shown by 60-day-old prenatally stressed rats, but not by 30- and 45-day old. Increased open arm ends exploration was shown by 45-day-old prenatally stressed males. Rearing behavior was found to increase with age, a phenomenon more pronounced in females. Additionally, at the younger ages prenatally stressed rats were heavier than controls, an effect which disappeared at young adulthood. In conclusion, anxiogenic prenatal stress effects in the elevated plus-maze could only be detected at early adulthood, not before. Nonetheless, at late adolescence (45 days of age) prenatal stress leaded to an anxiolytic-like effect which can be interpreted as increased risk-taking behavior.