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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.

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.

A seleção genética de comportamentos relacionados a um componente emocional do tipo ansiedade, associada ao estudo das possíveis influências desse traço sobre outros componentes emocionais como o medo, impulsividade e tarefas de aprendizagem aversiva, podem contribuir para o entendimento da ansiedade, bem como, fornecer uma base sólida para investigações neurofisiológicas, bioquímicas e moleculares da mesma. Assim, o presente trabalho selecionou duas linhagens de ratos da linhagem Wistar: uma com característica de alta exploração dos braços abertos do labirinto em cruz-elevado (Filô-HE) e outra com baixa exploração desses braços (Filô-LE), e comparou os efeitos da seleção dessas duas linhagens sobre comportamentos associados à ansiedade, medo, impulsividade e aprendizagem aversiva. Para isso, um lote de animais fornecidos pelo Biotério Central da USP-RP foram distribuídos em dois grupos distintos: (1) os que apresentavam maior exploração dos braços abertos do Labirinto em Cruz Elevado (LCE) - (Filô-HE) e (2) os com menor exploração desses braços (Filô-LE) ao longo de doze gerações (de S1 a S12, sem contar o grupo inicial do qual derivam as gerações). Na sexta, nona e décima segunda gerações, os animais foram submetidos aos seguintes testes comportamentais: LCE, Campo Aberto (somente na décima segunda), esquiva passiva, labirinto aquático de Morris (somente na décima segunda), sobressalto (que avalia a resposta de medo), e gradiente de aversão elevado (desenvolvido em nosso laboratório para medir impulsividade). Os dados indicaram a seleção de duas linhagens de ratos Wistar com características distintas para a exploração dos braços abertos do LCE. De um lado, foi selecionada a linhagem Filô-HE, que possui como principal característica, altos índices de exploração de ambientes que geralmente são evitados por outros animais. De outro, foi selecionada (até a oitava geração), a linhagem Filô-LE, que explorava muito pouco ambientes potencialmente aversivos. Tais resultados podem ser interpretados, respectivamente, como (a) baixa e alta frequência de comportamentos associados à ansiedade e medo, (b) baixa e alta frequência de comportamentos associados à impulsividade, (c) altos e baixos níveis de dificuldade na aprendizagem relacionada a memórias emocionais, ou (d) até mesmo o efeito conjunto de todas essas características comportamentais. Porém, para um conhecimento mais confiável, é necessário que os estudos supracitados passem a explorar aspectos neurofisiológicos, psicofarmacológicos, bioquímicos e genéticos, ampliando a investigação dos componentes emocionais selecionados. Bem como, promovendo um controle mais rígido dos traços selecionados e de suas relações com outros componentes que possam interferir na seleção The genetic selection of behaviors related to an anxiety-like emotional component, associated to the study of the possible influences of this trait over other emotional components like fear, impulsivity and aversive learning, can contribute to the understanding of anxiety as well as supply a solid basis for its neurophysiological, biochemical and molecular investigation. Thus, the present work selected two strains of Wistar rats: one characterized by a high exploration of the open arms of an elevated plus-maze (Filô-HE) and another with a low exploration of these arms (Filô-LE), and compared the effects of such a selection on behaviors associated with anxiety, fear, impulsivity and aversive learning. For this, a batch of animals supplied by the animal house of the USP at Ribeirão Preto were distributed into two distinct groups: (1) one presenting more exploration of the open arms of an elevated plus-maze (EPM) Filô-HE and (2) one with less open arm exploration Filô-LE along twelve generations (from S1 to S12, not counting the initial group from which all generations derived). On the sixth, ninth and twelfth generations the animals were submitted to the following behavioral tests: LCE, open-field (only in the twelfth generation), passive avoidance, Morris aquatic maze (only in the twelfth generation), startle (which evaluates a fear response), and elevated aversion gradient (developed in our laboratory to measure impulsivity). The data indicated a selection of two Wistar rats strains with distinct characteristics concerning the exploration of the open arms of and EPM. On the one hand, the strain Filô-HE was selected exhibiting as the main characteristic high measures of exploration of environments which are usually avoided by other animals. On the other hand, the strain Filô-LE was selected (up to the eighth generation) exhibiting very little exploration of potentially dangerous environments. These results may be interpreted as, respectively, (a) low and high frequency of behaviors associated to anxiety and fear, (b) low and high frequency of behaviors associated to impulsivity, (c) high and low levels of difficulty in learning related to emotional memories, or (d) even the joint effect of all these behavioral characteristics. However, for a more reliable knowledge it is necessary that the studies above begin to explore neurophysiological, psychopharmacological, biochemical and genetic aspects, broadening the investigation ot hte selected emotional components, as well as promoting a more strict control of the selected traits and of its relations with other components that may interfere in the selection

For decades, neuroscientists have been carrying out studies using animal models. Clinical professionals do not use the data originated from these studies because the interpretation of these studies may be a hard task for them. How data generated in studies with animals lead to practical use in psychological and psychiatric clinics is a common question in courses and meetings in this area of science. There are two typical mistakes often occurring in daily practice, namely, the transposition of data directly from laboratories to clinics or simply ignoring such data. In the first intance, the characteristics of species and the limits of these studies are ignored by therapists; in the second, the professionals do not read the studies because the issues are not familiar to them. We believe that this problem occurs because the concept of an animal model is not clear to many therapists. In this chapter we will discuss the concept of behavioral animal models, its limitations and possibilities of simulating a mental pathology; we show the differences between the use of animal models in behavioral sciences and in others fields of research in biomedical sciences; we discuss the criteria and processes of validation of an animal model; and finally, we give some advice to clinical professionals on the reading of animal models in the literature.

A validação de um modelo experimental para se investigar a ansiedade deve estar baseada inicialmente em uma validação comportamental paramétrica, que pressupõe a investigação de respostas relacionadas ao comportamento de defesa da espécie frente às situações aversivas presentes no ambiente de exposição. Assim, o presente trabalho validou a preferência claro/escuro como um modelo comportamental para o estudo da ansiedade no peixe Carassius auratus (peixe dourado). Foram avaliados os seguintes parâmetros: repetição das exposições ao aparato de teste em 5 sessões diárias, privação de comida, aquário enriquecido, troca de água do aquário antes dos testes, proporções diferentes do compartimento claro e escuro, e aquário com a metade do comprimento. Os dados indicaram que as reexposições aumentaram o número de cruzamentos, indicando que a mesma aumenta a exploração do aparato, atenuando os componentes aversivos do ambiente. Os peixes alojados no aquário enriquecido se locomoveram mais no aparato, e não apresentam preferência significativa por nenhum dos compartimentos (claro/escuro), evidenciando que a aversividade do ambiente claro foi atenuada pelo alojamento em um aquário enriquecido. Os animais privados de comida por 48 h não apresentaram diferenças estatisticamente significativas entre os tempos gastos nos compartimentos claro e escuro do aquário, indicando uma maior exploração do aparato em busca de alimento. Os animais que passaram pelo procedimento de troca de água do aquário de testes não apresentaram preferência por nenhum dos compartimentos, indicando que os peixes, provavelmente, liberam sinalizadores químicos na água que informam seus co-específicos sobre o potencial aversivo do ambiente. Quanto à proporção do aquário, os animais expostos ao aquário teste 75% claro e 25% escuro se locomoveram menos que os do aquário controle, indicando que uma maior área clara, pode levar a uma ampliação da aversividade nesse aquário. Quando os sujeitos foram submetidos ao aquário 75% escuro e 25% claro, a frequência de cruzamentos foi maior no grupo controle e, nesse caso, a maior locomoção pode estar associada a uma atenuação da aversividade do ambiente relacionada a presença de uma maior área escura. No aquário 87,5% claro e 12,5% escuro, os animais do grupo experimental não apresentaram diferenças estatisticamente significativas entre os tempos gastos em nenhum dos compartimentos do aquário, indicando que uma maior área clara (aversiva) altera a preferência por ambientes escuros. Os peixes expostos ao aquário com a metade do comprimento aumentaram sua locomoção, evidenciando que o comprimento do aparato interfere na aversividade do compartimento claro. De forma geral, os dados demonstraram que a manipulação de estímulos ambientais altera os padrões comportamentais exploratórios relacionados à preferência claro/escuro, os quais podem estar associados a diferentes comportamentos relacionados à ansiedade. The validation of an experimental model to investigate the anxiety must be initially based in a parametric behavioral validation, which involves investigating responses related to the defense behavior of the species to cope with aversive situations in the environment of exposure. Thus, the present study validated the light/dark preference as a behavioral model for the study of anxiety in Carassius auratus (goldfish). The following parameters were evaluated: repetition of exposures to the test apparatus in 5 daily sessions, food deprivation, enriched aquarium, exchange of the water of the test aquarium, different proportions of the light and dark compartments, and exposure to an aquarium with half the length of the control one. The data indicated that the repeated exposures increased the number of crossings, indicating that they increased the exploration of the apparatus by reducing the aversive components of the environment. The fish housed in the enriched aquarium displaced more in the test apparatus, showin no significant preference for either the dark or light side, indicating the aversion of the test environment was attenuated by the exposure to the enriched aquarium. 48-H food-deprived animals showed no statistically significant differences between the times spent in either side of the test aquarium, indicating more intense exploration of the test apparatus in search of food. The animals submitted to the procedure of exchanging the water of the test aquarium before testing did not show preference for either side, indicating that the fish probably release chemical signals in the water which inform their conspecifics of the potential aversiveness of environment. As to the proportion of the aquarium, animals exposed to the 75% light 25% dark test aquarium displaced less than the ones exposed to the control aquarium, indicating that a larger light area can lead to an increase in the aversiveness of the aquarium. When the subjects were exposed to the 75% dark 25% light aquarium, the frequency of crossings was greater for the control group and, in this case, the increased locomotion may be associated with an attenuation in the aversiveness of the environment related to the presence of a larger dark area. In the 87.5% light 12.5% dark aquarium, the animals in the experimental group did not present statistically significant differences between the times spent on either the light or dark sides of the apparatus, indicating that a larger light (aversive) area alters the preference for dark environments. The fishes exposed the aquarium with half the length increased their locomotion, indicating that the length of the apparatus interferes with the aversiveness of the light compartment. Overall, the data showed that manipulating environmental stimuli affects exploratory behavior patterns related to light/dark preference, which may be associated with different anxiety-related behaviors.

psychopathology, construct validity is usually enhanced by addressing theories from other fields in its nomological network. In the field of anxiety research, this construct is related to antipredator behavior, conserved across phylogeny in its functions and neural basis, but not necessarily on its topography. Even though the relations between behavioral models of anxiety and statements from behavioral ecology and evolutionary biology are commonly made in anxiety research, these are rarely tested, at least explicitly. However, in order to increase construct validity in experimental anxiety, testing predictions from those theories is highly desirable. This article discusses these questions, suggesting a few ways in which behavioral ecological and evolutionary hypotheses of anxiety-like behavior may be tested. Keywords: construct validity, animal models of anxiety, evolution, psychopathology.

Statins are inhibitors of the 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, thereby inhibiting cell synthesis of cholesterol and isoprenoids. Moreover, several studies have been evaluating pleiotropic effects of statins, mainly because they present neuroprotective effects in various pathological conditions. However, knowledge about behavioral effects of statins per se is relatively scarce. Considering these facts, we aimed to analyze behavioral responses of atorvastatin or simvastatin-treated mice in the open field test, elevated plus maze and object location test. Atorvastatin treatment for 7 consecutive days at 1mg/kg or 10mg/kg (v.o.) or simvastatin 10mg/kg or 20mg/kg enhanced cognitive performance in object location test when compared to control group (saline-treated mice). Simvastatin effects on mice performance in the object location test was abolished by post-training infusion of the beta-adrenoceptor antagonist propranolol. Atorvastatin and simvastatin did not change the behavioral response in open field and elevated plus-maze (EPM) tests in any of the used doses. These data demonstrate the positive effects of both statins in cognitive processes in mice, without any alteration in locomotor parameters in the open field test or anxiolytic-like behavior in EPM. In conclusion, we demonstrate that atorvastatin and simvastatin per se improve the cognitive performance in a rodent model of spatial memory and this effect is related to beta-adrenergic receptors modulation.

Os modelos animais etológicos têm recebido destaque por apresentarem vantagens como economia de tempo e metodologia simples, por se utilizarem de comportamentos naturais da espécie, não requerendo treinamento do animal. O Labirinto em Cruz Elevado (LCE) tem recebido destaque pelo frequente uso em testes de drogas ansiolíticas. O modelo é baseado num conflito entre exploração e esquiva de espaços abertos. O tempo de permanência de ratos nos braços abertos é considerado a principal medida de ansiedade do procedimento. Entretanto, o conflito causado no animal pelo teste cessa após um período curto de tempo e o animal apresenta uma preferência pelos braços fechados do aparato. Outra condição que a literatura apresenta é o efeito one-trial tolerance, que consiste em uma suspensão dos efeitos dos agentes ansiolíticos benzodiazepínicos em uma segunda exposição do animal ao LCE nas medidas comportamentais. O objetivo deste trabalho foi realizar uma alteração procedimental cujo propósito era o de prolongar o conflito que o teste apresenta. O procedimento consistiu em modificar o LCE adicionando luzes de intensidade média de 320 lux nos braços fechados e luz ambiente com média de 22 lux nos braços abertos. Foram testados 48 ratos em dois dias consecutivos. Os animais foram divididos randomicamente em dois grupos, um controle que recebeu intraperitonealmente salina e outro que recebeu midazolam (1 mg/Kg) diluído em salina. Os animais foram a seguir testados no LCE por cinco minutos (Sessão 1, S1). A Sessão 2 (S2) ocorreu no dia seguinte. Cada um dos grupos testados na S1 foi subdividido randomicamente em dois, de forma que metade recebeu salina e metade midazolam. Utilizou-se medidas comportamentais tradicionais no teste do LCE além de diversas medidas de grooming e posturas esticadas. Os resultados encontrados estão de acordo com a literatura da área, sendo que algumas medidas indicaram que o conflito permaneceu durante as duas exposições do animal ao labirinto, além de apontar para uma possível superação do efeito one-trial tolerance. Ethological animal models have been widely studied because they present advantages such as time saving and simple methodology by using natural animal training. The Elevated Plus Maze (EPM) has been frequently associated with the use of anxiolytic drug testing. The model is based on a conflict between exploration and avoidance of the open spaces. The time rats spend on the open arms is considered the main measure of anxiety in the procedure. However, the conflict caused in the animal by the test ceases after a short period of time, and the animal the shows a preference for the closed arms of the apparatus. Another condition presented in the literature is the one-trial tolerance effect, which consists of a suspension of effects of the benzodiazepine anxiolytic agents in a second exposure of the animal to the EPM on behavioral measures. The objective of this work was to perform a methodological change in order to prolong the conflict presented by the test. The procedure consisted of modifying the EPM by adding a 320-lux light in the closed arms, and a 22-lux ambient light in the open arms. We tested 48 mice on two consecutive days. The animals were randomly divided into two groups, a control, that received intraperitoneal saline, and another, that received midazolam (1 mg/kg). The animals were then tested in the EPM for five minutes (Session 1, S1). Session 2 (S2) occurred on the following day. Each of the groups tested in S1 was subdivided randomly into two, so that half received saline and half midazolam. Besides the traditional behavioral measures, grooming and stretching were also analyzed. The results are according to the literature of the area, since some measures indicated that the conflict remained during both trials, in addition to pointing to a possible overcoming of one-trial tolerance.