Os autores agradecem Drs. Kentaro Nagaoka (Universidade de Tóquio de Agricultura e Tecnologia) e Wataru Iio (Ibaraki) para a discussão útil. Esta pesquisa foi apoiada em parte por Ibaraki University Cooperação entre Agricultura e Ciências Médicas (IUCAM) (A MEXT, Japão) e do Projeto de Pesquisa sobre o Desenvolvimento de Produtos e Alimentos com benefícios de promoção da sanidade agropecuária (NARO) (O MAFF, Japão) .

Os estudos com animais foram aprovados pelo e cumpridas as orientações, tanto do Animal Care e Use Committee da Universidade de Ibaraki e do Ministério da Educação Cultura, Desporto, Ciência e Tecnologia (MEXT), Japão (Notificação No.71). Uma descrição completa do protocolo é demonstrado na Figura 1. 1. Aparelho <ol><li> Prepare dois tipos de gaiolas: uma única gaiola (largura [W] × profundidade [D] × altura [H] = 143 milímetros × 293 milímetros × 148 mm), e uma &quot;derrota sociais (SD)&quot; gaiola (W × D × H = 220 mm × 320 mm × 135 milímetros). </li><li> Como mostrado na Figura 2, a gaiola SD dividir em dois compartimentos com uma placa acrílica transparente divisor (5 mm de espessura) com 15 furos circulares (3 × 5 matriz: 8 mm de diâmetro). </li><li> Obter chips de madeira-de barbear feitos de abetos, purificada por dieta pelotas do alimento, e uma garrafa de água potável. Além disso, obter toalhas de papel, uma máscara e luvas de látex. </li&gt; <li> Para o teste de interação social, preparar uma arena de campo aberto (W × D × H = 40 centímetros x 40 centímetros x 40 cm) feitos de cloreto de polivinilo cinza, um peso de aço (190 g), e uma caixa de interação plástico (W × D × H = 10 centímetros x 10 centímetros × 13 cm; 100 g) com três janelas de malha de arame (W × D = 5 centímetros x 5 cm) (Figura 3). </li><li> Coloque uma câmara CCD (2,8-12 mm; F ​​= 1: 1,4; 1/3 polegada CCD) e um sistema de seguimento automático na sala de testes de comportamento na instalação para animais. Coloque uma prateleira apropriado para as gaiolas de rato na sala de testes de comportamento que se habituem os ratos para o ambiente da sala de teste durante pelo menos 30 min. </li></ol> 2. A habituação ao Meio Ambiente <ol><li> Use macho C57BL / 6JJmsSlc (B6) ratos que são 7 semanas de idade e do sexo masculino Slc: ICR (ICR) ratos que são mais de 5 meses de idade e entregar para a instalação de uma empresa de criação de animais. </li><li> Independentemente mover dois grupos de ratinhos B6 (n= 12 em cada grupo) para a instalação; utilize um grupo (B6 ratos screener) para o rastreio de ratinhos ICR agressivos e outro grupo (sujeito ratos B6) para o desenvolvimento do modelo sCSDS. </li><li> Introduzir ratinhos ICR (n = 12) para a instalação, a fim de rastrear seus comportamentos agressivos. </li><li> Casa os ratos individualmente em gaiolas individuais durante 1 semana sob um ciclo de luz-escuro de 12 h (cerca de 100 lux de luz fluorescente, luzes acesas às 08:00) com temperatura constante (cerca de 23 ° C) e os níveis de humidade (cerca de 40%) de modo a habituar-los para o ambiente. Particionar cada gaiola, colocando placas de plástico de cor branca, entre as gaiolas para que os ratos não são afetados pelos comportamentos dos ratos vizinhos. </li><li> Faça purificada por dieta grãos de ração e água de osmose reversa disponível ad libitum. Use AIN-93G Chow porque os ingredientes de outros peletes de dieta não purificada pode variar. </li><li> Medir o peso corporal, a ingestão de alimentos (FI), e ingestão de água (WI) dos ratinhos todos os dias. Calcule a bganho de peso Ody (BWG) desde o dia inicial. </li></ol> 3. Triagem de Aggressive ICR Mice <ol><li> Depois de habituação para 1 semana como mencionado acima, a tela de ratinhos ICR (residente), utilizando testes de residente-intrusão para um julgamento 3 min com ratos screener B6 (intruso; 8 semanas de idade) no período da tarde (14: 00-17: 00) sob a iluminação de cerca de 300 lux na sala de habitação. <ol><li> Especificamente, teste cada ratinho ICR para três ensaios por dia durante 5 dias consecutivos (15 ensaios no total) em relação a muitos ratos B6 diferentes quanto possível para encontrar que ratinhos ICR mostram alta agressividade para com os intrusos. Durante os testes, registrar o ataque latência e duração do comportamento agressivo (movimentos rápidos com ataque cortante). </li></ol></li><li> Identificar ratinhos ICR hiper-agressivo, verificando as feridas danos em ratos intrusos B6 após cada tentativa. </li><li> Calcular a proporção de ensaios em que a latência ataque é inferior a 30 segundos, como um primeiro índice de pontuação da agressão. </li><li> Calculate a proporção de ensaios em que a latência de ataque é menor do que 3 minutos, como um segundo índice de pontuação da agressão. </li><li> Avaliar os escores de agressividade desde o primeiro índice. Use o segundo índice quando o primeiro índice é igual. </li><li> Selecione os ratinhos ICR que tiveram pontuações mais altas de agressão sem comportamentos hiper-agressivo como ratinhos ICR agressivos. Use os ratinhos ICR agressivos repetidamente ao longo do próximo conjunto de experimentos até aproximadamente 12 meses de idade; no entanto, realizar o processo de triagem para ratos ICR agressivos como descrito acima antes de cada experimento para confirmar a agressividade dos ratinhos ICR. </li><li> Depois de cada sessão do rastreio, registrar o grau de lesão dos ratos screener B6. Se um mouse estiver ferido, isolar o mouse em uma única gaiola e observar o progresso, verificando seu ganho de peso corporal, a ingestão de alimentos e ingestão de água. No caso de feridas graves, que afectam a fisiologia e comportamento em ratinhos, sacrificar-los de acordo com I locaisOrientações ACUC. </li></ol> 4. sCSDS <ol><li> Introduzir ratinhos B6 sujeita 7 dias antes (dia -6) no dia da exposição inicial de stress (dia 1) e casa os ratos individualmente em gaiolas individuais para habituar-los para o ambiente. </li><li> Três dias antes do dia 1 (no dia -2), mover os ratinhos ICR agressivas para um compartimento de cada gaiola de SD, o qual é dividido por um divisor de acrílico para permitir que os ratinhos estabelecer seus territórios nas gaiolas SD (o mesmo número de sujeitos B6 ratos). </li><li> Separe as gaiolas SD usando placas de separação de cor branca, como no estado de gaiola única descrito acima. </li><li> Para ratinhos B6 não-esforçado de controlo, preparar gaiolas SD (metade do número de murganhos) para manter os ratos aos pares; colocar os dois ratos em cada compartimento dividido pelo divisor numa gaiola SD durante 10 dias. </li><li> Após as medições diárias BWG, FI, e WI no dia 1, os ratos local sujeito B6 em um dos compartimentos das gaiolas SD (home-gaiola do residente) no afternoon (14: 00-16: 00) com iluminação de cerca de 300 lux na sala de habitação e medir o tempo de contato físico a partir da primeira mordida ataque para completar o teste residente-intruso. </li><li> No dia 1, defina o tempo de contato físico para 5 min da primeira mordida ataque e ter observadores contar o número de picadas de ataque de ratos ICR que são direcionadas preferencialmente na parte de trás ou flancos do oponente como descrito por Miczek et al. 20 a determinar o grau de stress físico para ratinhos B6. </li><li> Após o tempo de contato físico, B6 resgate sujeitos, e verificar e registrar o seu estado de pele e feridas. Em seguida, colocar os ratinhos B6 para outro compartimento ao lado dos ratinhos ICR nas gaiolas SD até que a exposição ao stress físico no dia seguinte. </li><li> Porque o divisor de acrílico na gaiola SD é transparente e contém furos, expor ratinhos B6 sujeita a várias tensões emocionais, incluindo visual, auditiva e estímulos olfativos, dos ratinhos ICR no compartimento vizinhomento da gaiola SD durante 24 horas todos os dias. </li><li> Medir a BWG, FI, e WI de ratinhos B6 controle diário e, em seguida, colocá-los em cada compartimento nas gaiolas SD para 1 dia. </li><li> No dia 2, após as medições diárias de BWG, FI, e WI, introduzir ratinhos B6 sujeitos nos territórios de outros ratinhos ICR para expô-los ao estresse físico. </li><li> Defina a duração do contato físico em 4,5 min a partir da primeira mordida ataque no dia 2, e contar o número de picadas de ataque. </li><li> Mova ratinhos B6 controle em diferentes compartimentos e mudar as combinações de pares, a fim de embaralhar a colocação e parceiro dos pares de cada dia. </li><li> Diminuir a duração do contacto físico por 0,5 min por dia, de modo que a duração do dia em 10 é definida como 0,5 min a partir do primeiro ataque mordida. </li><li> No dia 7, substituir cerca de metade das aparas de madeira em todos os compartimentos nas gaiolas SD. </li><li> Após a exposição ao stress físico último no dia 10, mover-se ratinhos B6 sujeito em gaiolas individuais. Similar aos ratinhos B6 sujeitos, mova o controle camundongos B6 em gaiolas individuais no dia 10. </li><li> Se um rato ICR não mostra quaisquer picadas de ataque até 5 minutos em cada tentativa nos dias 1 a 10, pôr termo ao julgamento. Substituir o rato ICR com ratinhos ICR agressivos peças e realizar um julgamento alternativa para o mouse assunto B6. </li><li> Se um mouse estiver ferido, isolar o mouse em uma única gaiola e observar o progresso, verificando seu ganho de peso corporal, a ingestão de alimentos e ingestão de água. Siga as diretrizes IACUC para analgesia. No caso de ferimentos graves que afetam a fisiologia e comportamento em camundongos, sacrificá-los de acordo com as directrizes locais IACUC. </li></ol> 5. social Prevenção de teste (Teste de Interacção social) <ol><li> Realizar testes comportamentais para evitação social na parte da manhã (09: 00-12: 00) no dia 11. </li><li> Trinta minutos antes do ensaio, transferir as gaiolas dos ratinhos de controlo e sujeitos B6 para uma prateleira (luz fraca de menos de 1 lux) na comportamentalquarto sob uma iluminação de teste inferior a 20 lux para lhes permitir habituar ao ambiente. </li><li> Para reduzir os efeitos de ordem, alternadamente testar ratinhos de controlo e sujeitos B6. </li><li> Limpar uma arena de campo aberto (iluminada com uma luz de 20 lux no centro do campo) e uma caixa de interacção plástico usando toalhas de papel embebidos com etanol a 70% antes do teste comportamental para cada rato para remover fezes, urina, e todos os odores . </li><li> Coloque um rato desconhecido ICR (não usado como um agressor) a partir de uma única gaiola perto do aparelho de campo aberto. </li><li> Coloque um rato B6 para o campo aberto da caixa de interacção vago, como mostrado na Figura 4. Monitorar e analisar o seu comportamento durante 2,5 minutos usando o sistema de análise automática (descrito no passo 5.10). </li><li> Após o primeiro ensaio, remover o rato B6 do campo e colocá-lo na sua gaiola de origem. </li><li> Em seguida, coloque o macho desconhecido ICR rato (meta social) na caixa de interação e, em seguida, introduzir a B6rato para o campo aberto e monitorar o seu comportamento durante 2,5 min. </li><li> Após a segunda tentativa, retornar tanto a B6 e ratinhos ICR para as suas gaiolas, e limpar o domínio da interacção e a caixa, como mencionado acima. </li><li> Repita essas etapas para cada ratinho B6 para testar o seu comportamento social. Durante cada ensaio, ficha top-view de filmes usando uma câmera CCD. <ol><li> Após os testes comportamentais, calcular o tempo gasto na zona de interacção (em segundos) e na zona de canto (em segundos) para cada teste, como mostrado em Goto et ai. 13. Julgar a posição do rato com base no centro do rato. </li><li> Calcular a pontuação de interação social como 100 × (tempo de interação, com alvo) / (tempo de interação, sem alvo), seguindo os métodos publicados em Krishnan et al. 21. </li></ol></li></ol>

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Renal Physiol. 290 (2), F478-F485 (2006).</li><li>Flores, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Schedule-Induced+Polydipsia:+Searching+for+the+Endophenotype+of+Compulsive+Behavior.">Schedule-Induced Polydipsia: Searching for the Endophenotype of Compulsive Behavior.</a> World J. Neurosci. 4, 253-260 (2014).</li><li>Cryan, J. F., Dinan, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mind-altering+microorganisms:+the+impact+of+the+gut+microbiota+on+brain+and+and+behaviour.">Mind-altering microorganisms: the impact of the gut microbiota on brain and and behaviour.</a> Nat. Rev. Neurosci. 13 (10), 701-712 (2012).</li></ol>

Os autores declaram que não têm interesses financeiros concorrentes.

Houve diferenças definitivas no peso corporal entre sCSDS ratos e camundongos CSDS submetidos ao protocolo padrão CSDS (5 a 10 min de contato físico com os agressores por dia) 19. Os ratos mostraram aumento sCSDS BWG durante o período de stress, enquanto que os camundongos normais CSDS mostraram uma diminuição no peso corporal durante o período de estresse 21,22,23. Há uma diferença substancial entre os dois protocolos em termos da duração total do contato físico com os agressores duran...

Muitos tipos de eventos estressantes ocorrem ao longo da vida dos seres humanos. O estresse excessivo muitas vezes leva a conseqüências fisiológicas prejudiciais em humanos e animais. Nos seres humanos, eventos estressantes são os principais fatores de risco para a precipitação de transtornos psiquiátricos, como depressão 1. A Global Burden of Disease estudo indicaram que a depressão é uma das doenças mais incapacitantes em termos de anos de vida ajustados por incapacidade (DALY) e anos vividos com incapacidade 2. Além disso, a depressão representa a maior proporção de DALYs suicidas 3. Pessoas que sofrem de depressão têm dificuldade em gerir as suas vidas, e, como resultado, sua qualidade de vida, muitas vezes piora. Por conseguinte, existe uma forte necessidade de desenvolver agentes terapêuticos eficazes para melhorar a qualidade de vida desses pacientes. Muitos estudos têm sido realizados sobre os principais transtornos depressivos, e revelaram que a contribuição genética para a doença susceptibility é de cerca de 30-40%, a qual é explicada pelos efeitos de múltiplos loci de efeitos pequenos 4. Por causa dos mecanismos patogénicos subjacentes complexos depressão, a etiologia detalhada da doença permanece elusiva. Os relatórios clínicos indicam que existem subtipos de depressão, como melancólico e atípico depressão 5, que mostram reduzidas e aumento de peso corporal, respectivamente 6. Apesar de 25-30% e 15-30% dos pacientes com depressão representam características melancólicas e atípicos puros, respectivamente, a maioria deles tem características em comum de ambos os subtipos 7. Por conseguinte, a depressão major tem uma ampla variedade de sintomas. A fim de encontrar e desenvolver terapêuticas biomarcadores objectivas para os vários tipos de depressão em seres humanos, é importante estabelecer diversos modelos animais diferentes da depressão 8. Os modelos animais de depressão foram estabelecidas usando várias aproximações incluindo aprendidodesamparo, estresse leve crônico imprevisível e estresse social derrota crônica (CSDS) 9-12. Toyoda e colaboradores estabeleceram os modelos CSDS de ratos e ratinhos de 13-17, a fim de elucidar o metabolismo e comportamentos que estão associados com a depressão. Tendo em conta que modelos animais de depressão são avaliados pela validade de face 18, o contexto em que o modelo está estabelecido é importante. Além disso, Golden et al. 19 relataram os métodos para a criação de ratinhos CSDS em detalhe. Sabe-se que as deficiências de comportamento social de ratinhos CSDS pode ser recuperado por tratamento crónico, mas não pelo tratamento agudo com antidepressivos, e que eles compartilham sintomas semelhantes aos dos doentes com depressão, em termos da regulação da neurotrófico derivado do cérebro fator de seis. Goto et al. 13 previamente desenvolvido o subcrónica e leve estresse social derrota (sCSDS) modelo de mouse, modificando os métodosde Golden et al. 19. Os ratos sCSDS mostrar sintomas polydipsia- e hiperfagia-like seguintes ganhos de peso corporal e índice de água aumentou corpo 13. Neste relatório, o protocolo para o estabelecimento do modelo de rato sCSDS é fornecido e discutimos a utilidade deste modelo.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>single cage</td>
			<td>Charles River Laboratories Japan</td>
			<td></td>
			<td>width [W] &#215; depth [D] &#215; height [H] = 143 &#215; 293 &#215; 148 mm</td>
		</tr>
		<tr>
			<td>M cage</td>
			<td>Natsume Seisakusho</td>
			<td></td>
			<td>W &#215; D &#215; H = 220 &#215; 320 &#215; 135 mm</td>
		</tr>
		<tr>
			<td>Whiteflake</td>
			<td>Charles River Laboratories Japan</td>
			<td></td>
			<td>Wood-shaving chips made from spruce trees</td>
		</tr>
		<tr>
			<td>AIN-93G</td>
			<td>Oriental Yeast</td>
			<td></td>
			<td>purified-diet food pellets</td>
		</tr>
		<tr>
			<td>Kimtowel</td>
			<td>Nippon Paper Crecia Co.</td>
			<td></td>
			<td>Paper towels</td>
		</tr>
		<tr>
			<td>open-field arena</td>
			<td>O&#8217;Hara &#38; Co.</td>
			<td></td>
			<td>made of gray polyvinylchloride</td>
		</tr>
	</tbody>
</table>

a mouse model of subchronic and mild social defeat stress for understanding stress-induced behavioral and physiological deficits

Stressful life events often increase the incidence of depression in humans. To study the mechanisms of depression, the development of animal models of depression is essential. Because there are several types of depression, various animal models are needed for a deeper understanding of the disorder. Previously, a mouse model of subchronic and mild social defeat stress (sCSDS) using a modified chronic social defeat stress (CSDS) paradigm was established. In the paradigm, to reduce physical injuries from aggressors, the duration of physical contact between the aggressor and a subordinate was reduced compared to in the original CSDS paradigm. sCSDS mice showed increased body weight gain, food intake, and water intake during the stress period, and their social behaviors were suppressed after the stress period. In terms of the face validity of the stress-induced overeating and overdrinking following the increased body weight gain, the sCSDS mice may show some features related to atypical depression in humans. Thus, a mouse model of sCSDS may be useful for studying the pathogenic mechanisms underlying depression. This protocol will help establish the sCSDS mouse model, especially for studying the mechanisms underlying stress-induced weight gain and polydipsia- and hyperphagia-like symptoms.

Para monitorizar o grau de stress físico ao longo dos períodos de 10 dias, o número de picadas de ataque em ratos ICR foi contado manualmente por um investigador. A Figura 5A indica os valores individuais para o número de picadas de ataque recebidos. Houve grande variabilidade na fase inicial (cerca de 10-120 mordidas no dia 1), mas essa variabilidade foi reduzida na fase posterior (aproximadamente 5-20 mordidas no dia 10). Figura 5B indica que o número médio de ataque bites receb...

Watch this Scientific Journal Video about A Mouse Model of Subchronic and Mild Social Defeat Stress for Understanding Stress-induced Behavioral and Physiological Deficits at JoVE.com

A Mouse Model of Subchronic and Mild Social Defeat Stress for Understanding Stress-induced Behavioral and Physiological Deficits

Aqui, métodos para desenvolver um modelo de mouse subcrónica e leve estresse derrota social são descritas e utilizadas para investigar as características patogênicas de depressão, incluindo hyperphagia- e sintomas polidipsia-like seguintes aumento do peso corporal.

Um modelo de mouse subcrónica e Mild Derrota social Estresse para Compreender déficits comportamentais e fisiológicas induzida por estresse

Stressful life events often increase the incidence of depression in humans. To study the mechanisms of depression, the development of animal models of ...

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Research

JoVE Journal

Behavior

19.4K visualizações.  Ibaraki University.  Aqui, métodos para desenvolver um modelo de mouse subcrónica e leve estresse derrota social são descritas e utilizadas para investigar as características patogênicas de depressão, incluindo hyperphagia- e sintomas polidipsia-like seguintes aumento do peso corporal. 

Vídeo: A Mouse Model of Subchronic and Mild Social Defeat Stress for Understanding Stress-induced Behavioral and Physiological Deficits

The Resident-intruder Paradigm: A Standardized Test for Aggression, Violence and Social Stress

This video publication explains in detail the experimental protocol of the resident-intruder paradigm in rats. This test is a standardized method to measure offensive aggression and defensive behavior in a semi natural setting. The most important behavioral elements performed by the resident and the intruder are demonstrated in the video and illustrated using artistic drawings. The use of the resident intruder paradigm for acute and chronic social stress experiments is explained as well. Finally, some brief tests and criteria are presented to distinguish aggression from its more violent and pathological forms.

This video shows the resident-intruder paradigm in rats. This test is a standardized method to measure offensive aggression, defensive behavior and violence in a semi-natural setting. The use of the paradigm for social stress experiments is explained as well.

O Paradigma Residente-intruder: um teste padronizado para Agressão, Violência e Estresse social

This video publication explains in detail the experimental protocol of the resident-intruder paradigm in rats. This test is a standardized method to ...

Comportamento

Using Chronic Social Stress to Model Postpartum Depression in Lactating Rodents

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals and humans. However, many animal models of depression were developed in males and are not applicable or effective in studies of postpartum females. Recent studies have reported significant effects of chronic social stress during lactation, an ethologically relevant and effective stressor, on maternal behavior, growth, and behavioral neuroendocrinology. This manuscript will describe this chronic social stress paradigm using repeated exposure of a lactating dam to a novel male intruder, and the assessment of the behavioral, physiological, and neuroendocrine effects of this model. Chronic social stress (CSS) is a valuable model for studying the effects of stress on the behavior and physiology of the dam as well as her offspring and future generations. The exposure of pups to CSS can also be used as an early life stress that has long term effects on behavior, physiology, and neuroendocrinology.

This article describes the use of chronic resident intruder social stress as an ethologically relevant paradigm to model postpartum depression and anxiety in lactating rodents.

Usando Social Stress Crônico ao Modelo depressão pós-parto em lactação Roedores

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals ...

Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several independent laboratories have demonstrated robust alterations in the social behavior of rodents exposed to alcohol during brain development across a wide range of exposure durations, timing, doses, and ages at the time of behavioral quantification. Prior work from this laboratory has identified reliable alterations in specific forms of social interaction following moderate prenatal alcohol exposure (PAE) in the rat that persist well into adulthood, including increased wrestling and decreased investigation. These behavioral alterations have been useful in identifying neural circuits altered by moderate PAE1, and may hold importance for progressing toward a more complete understanding of the neural bases of PAE-related alterations in social behavior. This paper describes procedures for performing moderate PAE in which rat dams voluntarily consume ethanol or saccharin (control) throughout gestation, and measurement of social behaviors in adult offspring.

The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood.

Moderado pré-natal álcool Exposição e quantificação do comportamento social em ratos adultos

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

A Dimensão Social do Stress: Manipulações experimentais de apoio social e de identidade social no Trier Social Stress Teste

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents

Chronic, unresolved stress is a major risk factor for the development of clinical depression. While many preclinical models of stress-induced depression have been reported, the unpredictable chronic mild stress (UCMS) protocol is an established translationally-relevant model for inducing behavioral symptoms commonly associated with clinical depression, such as anhedonia, altered grooming behavior, and learned helplessness in rodents. The UCMS protocol also induces physiological (e.g., hypercortisolemia, hypertension) and neurological (e.g., anhedonia, learned helplessness) changes that are clinically associated with depression. Importantly, UCMS-induced depressive symptoms can be ameliorated through chronic, but not acute, treatment with common SSRIs. As such, the UCMS protocol offers many advantages over acute stress protocols or protocols that utilize more extreme stressors. Our protocol involves randomized, daily exposures to 7 distinct stressors: damp bedding, removal of bedding, cage tilt, alteration of light/dark cycles, social stresses, shallow water bath, and predator sounds/smells. By subjecting rodents 3-4 hr daily to these mild stressors for 8 weeks, we demonstrate both significant behavioral changes and poor health outcomes to the cardiovascular system. This approach allows for in-depth interrogation of the neurological, behavioral, and physiological alterations associated with chronic stress-induced depression, as well as for testing of new potential therapeutic agents or intervention strategies.

The unpredictable chronic mild stress (UCMS) protocol is a validated method for studying behavioral and physiological changes associated with chronic stress and depressive symptoms. Eight weeks of imposition of the UCMS protocol induces behavioral changes and poor health outcomes in rodents of either gender.

Uma crônica Protocolo de estresse leve Imprevisível para sintomas depressivos Instigando, mudanças de comportamento e resultados negativos de saúde em Roedores

Chronic, unresolved stress is a major risk factor for the development of clinical depression. While many preclinical models of stress-induced depression ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, few studies have investigated the long-term effects of injury in rodent models of TBI. Shown here are behavioral measures that are well-established in TBI research for times early after injury, such as two weeks, until two months. Some of these methods have previously been used at later times after injury, up to one year, but by very few laboratories. The methods demonstrated here are a short neurological assessment to test reflexes, a Beam-Balance to test balance, a Beam-Walk to test balance and motor coordination, and a working memory version of the Morris water maze that can be sensitive to deficits in reference memory. Male rats were handled and pre-trained to neurological, balance, and motor coordination tests prior to receiving parasagittal fluid percussion injury (FPI) or sham injury. Rats can be tested on the short neurological assessment (neuroscore), the beam-balance, and the Beam-Walk multiple times, while testing on the water maze can only be done once. This difference is because rats can remember the task, thus confounding the results if repeated testing is attempted in the same animal. When testing from one to three days after injury, significant differences are detected in all three non-cognitive tasks. However, differences in the Beam-Walk task were not detectable at later time points (after 3 months). Deficits were detected at 3 months in the Beam-Balance and at 6 months in the neuroscore. Deficits in working memory were detected out to 12 months after injury, and a deficit in a reference memory first appeared at 12 months. Thus, standard behavioral tests can be useful measures of persistent behavioral deficits after FPI.

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after injury.

Detecção de défices comportamentais em ratos após lesão cerebral traumática

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by many patients prompts the need to develop new therapeutic alternatives and to better understand the etiology of the disorder. Pre-clinical models with translational merits are rudimentary for this task. Here we present a protocol for the unpredictable chronic mild stress (UCMS) method in mice. In this protocol, adolescent mice are chronically exposed to interchanging unpredictable mild stressors. Resembling the pathogenesis of depression in humans, stress exposure during the sensitive period of mice adolescence instigates a depressive-like phenotype evident in adulthood. UCMS can be used for screenings of antidepressants on the variety of depressive-like behaviors and neuromolecular indices. Among the more prominent tests to assess depressive-like behavior in rodents is the sucrose preference test (SPT), which reflects anhedonia (core symptom of depression). The SPT will also be presented in this protocol. The ability of UCMS to induce anhedonia, instigate long-term behavioral deficits and enable reversal of these deficits via chronic (but not acute) treatment with antidepressants strengthens the protocol&#39;s validity compared to other animal protocols for inducing depressive-like behaviors.

Here we present the unpredictable chronic mild stress protocol in mice. This protocol induces a long-term depressive-like phenotype and enables to assess the efficacy of putative antidepressants in reversing the behavioral and neuromolecular depressive-like deficits.

O protocolo de estresse suave crônico imprevisível para induzir Anhedonia em ratos

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by ...

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders such as post-traumatic stress disorder (PTSD). To understand the biological mechanisms of fear dysregulation in PTSD, it is important to start with a valid animal model of the disorder. This protocol describes the methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of PTSD, in both rats and mice. SEFL was developed to recapitulate critical aspects of PTSD, including long-term sensitization of fear learning caused by an acute stressor. SEFL uses aspects of Pavlovian fear conditioning but produces a distinct and robust sensitized fear response far greater than normal conditional fear responses. The trauma procedure involves placing a rodent in a conditioning chamber and administering 15 unsignaled shocks randomly distributed over 90 minutes (for rat experiments; for mouse experiments, 10 unsignaled shocks randomly distributed over 60 minutes are used). On day 2, rodents are placed in a novel conditioning context where they receive a single shock; then, on day 3 they are placed back in the same context as on day 2 and tested for changes in freezing levels. Rodents that previously received the trauma display enhanced levels of freezing on the test day compared to those that received no shocks on the first day. Thus, with this model, a single highly stressful experience (the trauma) produces extreme fear of the stimuli associated with the traumatic event.

Here we describe the detailed methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of post-traumatic stress disorder, in both rats and mice. The model utilizes aspects of Pavlovian fear conditioning and freezing as an index of enhanced fear in rodents.

Aprendizagem avançada stress medo, um modelo robusto de roedor do transtorno de estresse pós-traumático

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

Um guia Visual para estudar as defesas comportamentais para patógeno ataca em formigas cortadeiras

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...

Nest Building Behavior as an Early Indicator of Behavioral Deficits in Mice

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents synthetic and/or natural materials (such as twine, tissue, cotton, paper, and hay) as a gauge of their overall well-being and as an ancillary assessment to predict the possible decline in cognition. Typically, changes in nesting behaviors, such as failure to create a nest, indicate a change in health or welfare. In addition, nesting behavior is sensitive to many environmental and physiological challenges, as well as many genetic mutations underlying pathological disease states. The following protocol describes a nesting behavior paradigm that explores the usage of four types of nesting material. In addition, the protocol utilizes intraclass correlations to demonstrate that inter-rater reliability is higher when nests are constructed out of shredded paper compared to other common nesting materials such as cotton squares, paper twists, and soft cob bedding. The chosen methodology and statistical considerations (i.e., intraclass correlation) for this assay may be of interest for those conducting experiments assessing the quality of living of mice.&#160;

Here, we present a protocol to quantify nest building behavior in mice, which is known to be impaired in several neurological disorders and diseases. This protocol examines the utility of four materials and offers the opportunity to quantify the rater agreement in scoring, improving the validity and reliability of the assay.