Os autores desejam agradecer Sr. Akihiro Hirano por sua assistência em produzir o aparelho personalizado. Os autores gostaria de agradecer o Sr. Kosuke Takahashi e Sr. Shingo Nakajima para sua assistência técnica. Este trabalho foi apoiado pela pesquisa sobre o risco das substâncias químicas, saúde e bolsas de investigação do laboratório de Ciências, Ministério da saúde, trabalho e bem-estar, Japão (H25-KAGAKU-IPPANN-003) e A concessão do projeto de pesquisa por Instituto de ciência e tecnologia de Meiji University, Japão.

Todos os procedimentos experimentais descritos aqui foram aprovados pelo cuidado institucional do Animal e uso Comitê da Universidade de Meiji (IACUC 14-0002(1)). Animais utilizados nos resultados representativos foram preparados como segue: ratos machos Sprague-Dawley foram comprados de uma empresa de animais de laboratório e alojados em grupos de 4 animais por jaula de rato padrão (W200 × D410 × H250 mm) pelo menos uma semana antes do início da experimento comportamental. O quarto de alojamento dos animais foi mantido nas condições normais de ambientes para a luz (12:12 horário de claro/escuro com luzes acesas no 10:00 h), temperatura (25,0 ± 0,5 ° C), umidade (55 ± 10%) e ventilação (10 vezes / h), e os animais tinham acesso ad libitum a comida ( Oriental fermento co. Ltd., Tóquio, Japão) e água. Os animais foram mantidos em uma gaiola de rato padrão (W410 x D250 x H200 mm, Toyoriko co., Ltd., Aichi, Japão) com papel do fundamento (alfa-dri, LSG Corporatin, Tóquio, Japão).
1. elevada e mais teste de labirinto
<ol><li>Configurar o plus elevado labirinto aparelhos em um contêiner personalizado (W1, 500 × D1, 500 × H2, 000mm) (figura 1A) com um quadro metálico em forma de mais 500 mm acima do piso para apoiar a execução braços forma (figura 1B), iluminação no teto para contêiner Ilumine a superfície do labirinto para a intensidade da luz exigida (Figura 1) e uma câmera de vídeo digital no teto centro para permitir o monitoramento de vídeo ao vivo dos testes comportamentais (Figura 1).</li>
	<li>Os braços (L500 x W100 mm) na estrutura em forma de mais e em seguida sujeite a plataforma quadrado central (W100 x D100 mm) no centro dos quatro braços (Figura 1 -F). Nota: Em nosso laboratório, temos dois tipos de braços abertos (L500 x W100 mm) e dois tipos de braços fechados (L500 x x H450 W100 mm) (Figura 1). Os braços fechados têm paredes ou transparentes ou opacas (Figura 1), e os braços abertos são projetados com ou sem uma borda curta (H5 mm, Figura 1E). A plataforma quadrado central (W100 × D100 mm) é colocada no centro dos quatro braços (Figura 1 -F). Assim, as seguintes quatro combinações de braços podem ser preparadas de acordo com finalidade experimental: abre braços com bordas + braços fechados com paredes transparentes (lajes/transparente), braços abertos, com bordas + braços fechados com paredes opacas (lajes/opaco), de braços abertos sem bordas + braços fechados com paredes transparentes (não-bordas/transparente) e braços abertos sem bordas + braços fechados com opaco paredes (não-bordas/opaco). A superfície da pista de braço é preta, para permitir melhor animal rastreamento pelo software de análise do comportamento automatizado, como descrito na secção 2, análise comportamental.</li>
	<li>Ajuste a intensidade de iluminação na superfície da parte distal da extremidade de cada braço e uma plataforma central para aproximadamente 100 Lux.</li>
	<li>Antes de cada ensaio, limpe todo o aparelho com solução de etanol 70% e limpe com água destilada.</li>
	<li>Clique no botão"gravar" no gravador de disco rígido para iniciar a câmera de vídeo digital que vai gravar o comportamento do rato para o vídeo descrito na secção 2, análise comportamental de análise de imagem.</li>
	<li>Coloque um animal na plataforma central, enfrentando um braço aberto e permitir a livre exploração em cima da aparelho por 5 min.</li>
	<li>Clique no botão"parar" no gravador de disco rígido, para terminar a gravação.</li>
	<li>Conte o número de fezes e micções após retornar o animal testado para o quarto de alojamento dos animais.</li>
	<li>Repita da etapa 1.4 até o final do experimento. Nota: Uma execução de teste seco é sempre efectuada antes de iniciar cada experimento para verificar que todas as configurações experimentais estão corretamente configuradas antes de quaisquer animais experimentais estão expostos a condições experimentais. Caso contrário, alguns animais, especialmente o primeiro animal pode experimentar diferentes condições ambientais (por exemplo, ruídos, possivelmente induzidos pelo experimentador durante pistas de manipulação e/ou animal, eventualmente deixadas pelo animal previamente testado). Nota: Os experimentadores estão perto do recipiente quando um animal está sendo testado. Assim, mesmo que os experimentadores não são visíveis o rato pode ainda ser capaz de senti-los através do cheiro e/ou som.</li>
</ol>2. análise comportamental
Nota: Análise comportamental é realizado pela observação manual e automatizada de software de análise.
<ol><li>
Observação do manual
	<ol>
<li>Depois que o animal é colocado no peitoril, observar o seguinte: número de entradas (uma entrada para um braço é definida como qualquer instância quando todas as 4 patas no braço), o tempo gasto em cada braço, e número de esticada-atendente Postures posturas enfrentando um braço aberto , ao afastar a plataforma central para um braço fechado.</li>
		<li>Medir a duração e a frequência dos comportamentos definidos acima pelo cronômetro e contagem de 5 min, a partir de quando as portas do recipiente estão fechadas.</li>
	</ol>
</li>
	<li>
Análise automatizada
	<ol>
<li>Usando o programa de software disponíveis, gravar o seguinte: a distância percorridas, tempo gasto em diferentes posições de braços abertos (segmentos proximais, médios e distais da plataforma central), e uma análise de minuto a minuto do tempo gasto em aberto e fechado braços.</li>
		<li>Clique no botão"Iniciar" do software análise automatizada ao mesmo tempo, como a observação manual começa (a gravação irá parar automaticamente se a duração de gravação é definido). Nota: O software de análise comportamental utilizado neste estudo rastreia o animal através do seu contraste com o campo de fundo; o animal, que é atribuído a um tamanho fixo, é identificado como o objeto mais brilhante e a locomoção de seu ponto central, portanto, pode ser usada para rastreamento de animais.</li>
	</ol>
</li>
</ol>

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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+open+arm+ledges+and+maze+experience+in+the+elevated+plus-maze.">The influence of open arm ledges and maze experience in the elevated plus-maze.</a> Pharmacol Biochem Behav. 54 (1), 31-40 (1996).</li><li>Horii, Y., Kawaguchi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Higher+detection+sensitivity+of+anxiolytic+effects+of+diazepam+by+ledge-free+open+arm+with+opaque+walled+closed+arm+elevated+plus+maze+in+male+rats.">Higher detection sensitivity of anxiolytic effects of diazepam by ledge-free open arm with opaque walled closed arm elevated plus maze in male rats.</a> Behav Brain Res. 294, 131-140 (2015).</li><li>Santos, R. 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Os autores não têm nada para divulgar.

À medida que aumenta o número de pacientes que sofrem de transtornos psiquiátricos, incluindo transtorno de ansiedade, é importante desenvolver estudos de ponta para melhorar a compreensão do mecanismo básico de ansiedade. Para conseguir isso, é também importante otimizar os procedimentos experimentais. Portanto, para melhorar a aplicação da elevada além de teste de labirinto, temos demonstrado o efeito das características de braço em resultados experimentais utilizando ratos.
A e...

O número de pacientes que sofrem de transtornos do humor, incluindo transtorno de ansiedade e depressão, está aumentando. Embora o desenvolvimento destes transtornos envolve fatores genéticos e ambientais, os mecanismos de patogenicidade detalhados ainda não estão totalmente compreendidos1. Portanto, pesquisa deve centrar-se em desvendar os mecanismos subjacentes de transtornos do humor, bem como o desenvolvimento de medicamentos eficazes que têm menos efeitos colaterais.
Testes comportamentais para animais de laboratório são úteis para investigar a função superior do cérebro, incluindo a emoção. Como comportamento animal é altamente variável, resultados experimentais diferem frequentemente entre laboratórios, mesmo se uma estirpe de pura idêntica foi usado2. Assim, os pesquisadores também precisam entender as características básicas dos dispositivos experimentais adoptarem a fim de fornecer resultados fiáveis, precisos no laboratório.
A elevada além de teste de labirinto foi originalmente criado para examinar a reatividade emocional em roedores3,4. Todo o aparelho é normalmente localizado 500 mm acima do piso e consiste de dois abertos e dois braços fechados que fazem de uma forma mais. Medições de exploração do braço (incluindo a duração e a frequência de entradas em cada braço), distância viajados e especificados comportamentos como postura esticada-atendente de animais sujeitos são registados durante os períodos de observação. Por essas medidas, estima-se como um resultado experimental animal ansiedade.
A elevada além de teste de labirinto é usado em todo o mundo, mas nem todos os laboratórios usam o mesmo aparelho design5,6, e estas diferenças podem afectar os resultados experimentais. Por exemplo, diferenças incluem a presença ou ausência de saliências ao longo de braços abertos e/ou o uso de paredes transparentes ou opacos para braços fechados5,6. As bordas ao longo de braços abertos e o uso de paredes transparentes para braços fechados foram desenvolvidos para evitar animais cair o aparelho7e para facilitar as observações comportamentais dentro os braços fechados, respectivamente de8,9. Embora tenha havido alguns estudos que examinaram as diferenças de estrutura de braço como individuais podem afectar a capacidade de detectar animais ansiedade1,2,5,10,11 , os efeitos combinados de open (com/sem borda curta) e características de braço fechado (transparente/opaco paredes) sobre o comportamento do animal ainda não tenha sido investigada.
Num relatório recente, discutimos o efeito destas combinações de designs de braço no comportamento de ansiedade em ratos12. Em breve, bordas de braço aberto aumentaram significativamente comportamento de ansiedade em ratos, enquanto braços transparentes também podem ter diminuído esse comportamento. Além disso, verificamos a maior sensibilidade de detecção do efeito de um medicamento ansiolítico em uma combinação de braço aberto não-ledged + opaco braço fechado murado em comparação com uma combinação de ledged braço aberto + braço fechado murado transparente. Neste relatório, podemos apresentar nosso protocolo para a elevada mais labirinto teste, juntos com a discussão dos principais resultados do relatório anterior. Acreditamos que este relatório fornecerá informações úteis para aqueles que têm empregado ou quem planeja usar a elevada mais de labirinto em seus estudos.

<table><tbody><tr><td>Customized container (W1500 &amp;times; D1500 &amp;times; H2000 mm)</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Lighting</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Elevated plus maze apparatus</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Digital video camera</td><td>Watec</td><td>WAT-250D2</td><td>Any suitable video camera can be attached to a hard disk recorder for recording tracking profiles.</td></tr><tr><td>Hard disk recorder</td><td>TOSHIBA</td><td>RD-R200</td><td>Any suitable hard disk recorder can be attached to the digital video camera.</td></tr><tr><td>Sprague-Dawley rats</td><td>Charles River Laboratories Japan, Inc.</td><td>NA</td><td>Any suitable rats can be used for your study</td></tr><tr><td>Standard rat cage</td><td>Toyoriko</td><td>TP-105</td><td>Any suitable rat cage be used for your study</td></tr><tr><td>Instrument to measure light intensity</td><td>AS ONE</td><td>1-5818-02</td><td>Any suitable instrument to measure light intensity</td></tr><tr><td>EthoVision XT version 8.0</td><td>Noldus</td><td>NA</td><td>Any suitable software to measure animal behavior</td></tr><tr><td>Diazepam</td><td>Taiyo Pharmaceutical Industry</td><td>1124402A2061</td><td>Any suitable drug can be used for your study</td></tr><tr><td>Tween 80</td><td>Wako Pure Chemical Industries</td><td>9005-65-6</td><td>Any suitable drug can be used for your study</td></tr></tbody></table>

testing animal anxiety in rats: effects of open arm ledges and closed arm wall transparency in elevated plus maze test

A elevada além de labirinto é um teste comportamental para avaliar ansiedade animal em roedores. Embora este teste é aplicado extensamente no campo da ciência comportamental, resultados conflitantes são fornecidos frequentemente de laboratórios diferentes. Para identificar razões para os diferentes resultados, enfocamos anteriormente características de braço, que diferem entre laboratórios, mais notavelmente a presença/ausência de bordas para os lados de braços abertos e a opacidade/transparência das paredes de braço fechado. Em um relatório anterior, usamos um contêiner personalizado projetado para comparar o comportamento de ratos em diferentes combinações de desenhos de braço abertos e fechados, em condições experimentais idênticas e mostrou diferenças de características de braço interferem experimental resultados. Em breve, bordas de braço aberto aumentaram significativamente comportamento de ansiedade em ratos, enquanto braços transparentes também podem ter diminuído esse comportamento. Além disso, verificamos uma maior sensibilidade de detecção do efeito de um medicamento ansiolítico em uma combinação de braço aberto não-ledged + opaco braço fechado murado em comparação com uma combinação de ledged braço aberto + braço fechado murado transparente. Neste relatório, nós apresentamos o nosso protocolo para a elevada plus teste de labirinto, juntamente com a discussão dos principais resultados do relatório anterior e nossa experiência experimental. Acreditamos que este relatório fornecerá informações úteis para pesquisadores que têm empregado ou quem planeja usar a elevada mais de labirinto em seus estudos.

No estudo anterior, comparamos os efeitos combinados de bordas de braço aberto e transparência das paredes de braço fechado sobre os comportamentos exploratórios no labirinto12. Ratos, significativamente, gastou muito menos tempo em braços abertos sem bordas do que de braços abertos, com bordas (F [1, 60] = 13.49, P = 0,0005) (Figura 2A). Resultados semelhantes foram encontrados também para o número de entradas nos bra...

Watch this Scientific Journal Video about Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test at JoVE.com

Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

Testes Animal ansiedade em ratos: efeitos de bordas de braço aberto e fechado do braço de parede transparência em elevados além de teste de labirinto

Ansiedade-como o comportamento dos ratos testados da elevada mais labirinto altamente varia de acordo com o ambiente de teste. Aqui nós mostramos como configurações do aparelho, especialmente a estrutura de braço, afetam o comportamento no labirinto, e explicamos um protocolo confiável para detectar os efeitos dos ansiolíticos. Este relatório irá contribuir para a investigação de ansiedade.

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral ...

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Universidad San Sebastian

Research

JoVE Journal

Behavior

13.5K visualizações.  Meiji University. Ansiedade-como o comportamento dos ratos testados da elevada mais labirinto altamente varia de acordo com o ambiente de teste. Aqui nós mostramos como configurações do aparelho, especialmente a estrutura de braço, afetam o comportamento no labirinto, e explicamos um protocolo confiável para detectar os efeitos dos ansiolíticos. Este relatório irá contribuir para a investigação de ansiedade.

Vídeo: Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

The Successive Alleys Test of Anxiety in Mice and Rats

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

O Becos Teste sucessiva de ansiedade em camundongos e ratos

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

Comportamento

Barnes Maze Testing Strategies with Small and Large Rodent Models

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land (Barnes) mazes. Improved performance over sessions or trials is thought to reflect learning and memory of the escape cage/platform location. Considered less stressful than water mazes, the Barnes maze is a relatively simple design of a circular platform top with several holes equally spaced around the perimeter edge. All but one of the holes are false-bottomed or blind-ending, while one leads to an escape cage. Mildly aversive stimuli (e.g.&#160;bright overhead lights) provide motivation to locate the escape cage. Latency to locate the escape cage can be measured during the session; however, additional endpoints typically require video recording. From those video recordings, use of automated tracking software can generate a variety of endpoints that are similar to those produced in water mazes (e.g.&#160;distance traveled, velocity/speed, time spent in the correct quadrant, time spent moving/resting, and confirmation of latency). Type of search strategy (i.e.&#160;random, serial, or direct) can be categorized as well. Barnes maze construction and testing methodologies can differ for small rodents, such as mice, and large rodents, such as rats. For example, while extra-maze cues are effective for rats, smaller wild rodents may require intra-maze cues with a visual barrier around the maze. Appropriate stimuli must be identified which motivate the rodent to locate the escape cage. Both Barnes and water mazes can be time consuming as 4-7 test trials are typically required to detect improved learning and memory performance (e.g.&#160;shorter latencies or path lengths to locate the escape platform or cage) and/or differences between experimental groups. Even so, the Barnes maze is a widely employed behavioral assessment measuring spatial navigational abilities and their potential disruption by genetic, neurobehavioral manipulations, or drug/ toxicant exposure.

The dry-land Barnes maze is widely used to measure spatial navigation ability in response to mildly aversive stimuli. Over consecutive days, performance (e.g.&#160;latency to locate escape cage) of control subjects improves, indicative of normal learning and memory. Differences between rats and mice necessitate apparatus and methodology changes that are detailed here.

Barnes Estratégias Teste Labirinto com pequenas e grandes roedores Modelos

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land ...

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

A utilização do campo aberto Maze medir Locomotor e Ansiedade-like Comportamento no Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For many patients afflicted with CNS and non-CNS malignancies, radiotherapy and chemotherapy offer the best options for disease control. These treatments however come at a cost, and nearly all cancer survivors (~11 million in the US alone as of 2006) incur some risk for developing cognitive dysfunction, with the most severe cases found in patients subjected to cranial radiotherapy (~200,000/yr) for the control of primary and metastatic brain tumors1. Particularly problematic are pediatric cases, whose long-term survival plagued with marked cognitive decrements results in significant socioeconomic burdens2. To date, there are still no satisfactory solutions to this significant clinical problem.
We have addressed this serious health concern using transplanted stem cells to combat radiation-induced cognitive decline in athymic rats subjected to cranial irradiation3. Details of the stereotaxic irradiation and the in vitro culturing and transplantation of human neural stem cells (hNSCs) can be found in our companion paper (Acharya et al., JoVE reference). Following irradiation and transplantation surgery, rats are then assessed for changes in cognition, grafted cell survival and expression of differentiation-specific markers 1 and 4-months after irradiation. To critically evaluate the success or failure of any potential intervention designed to ameliorate radiation-induced cognitive sequelae, a rigorous series of quantitative cognitive tasks must be performed. To accomplish this, we subject our animals to a suite of cognitive testing paradigms including novel place recognition, water maze, elevated plus maze and fear conditioning, in order to quantify hippocampal and non-hippocampal learning and memory. We have demonstrated the utility of these tests for quantifying specific types of cognitive decrements in irradiated animals, and used them to show that animals engrafted with hNSCs exhibit significant improvements in cognitive function3.
The cognitive benefits derived from engrafted human stem cells suggest that similar strategies may one day provide much needed clinical recourse to cancer survivors suffering from impaired cognition. Accordingly, we have provided written and visual documentation of the critical steps used in our cognitive testing paradigms to facilitate the translation of our promising results into the clinic.

Cognitive impairment resulting from the radiotherapeutic management of brain tumors represents a clinically intractable condition that adversely impacts quality of life. The capability to critically evaluate potential interventions for ameliorating radiation-induced cognitive decrements ultimately depends on the capability to undertake rigorous quantitative assessments of cognition.

Diminui quantificar cognitivos causados ​​pela radioterapia craniana

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For ...

Medicina

A New Method for Inducing a Depression-Like Behavior in Rats

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is no existing format for studying the mechanism of action, prevention, containment, and treatment of contagious depression. The purpose of this study, therefore, was to establish the first animal model of contagious depression.
Healthy rats can contract depressive behaviors if exposed to depressed rats. Depression is induced in rats by subjecting them to several manipulations of chronic unpredictable stress (CUS) over 5 weeks, as described in the protocol. A successful sucrose preference test confirmed the development of depression in the rats. The CUS-exposed rats were then caged with na&#239;ve rats from the contagion group (1 na&#239;ve rat/2 depressed rats in a cage) for an additional 5 weeks. 30 social groups were created from the combination of CUS-exposed rats and na&#239;ve rats.
This proposed depression-contagion protocol in animals consists mainly of cohabiting CUS-exposed and healthy rats for 5 weeks. To ensure that this method works, a series of tests are carried out - first, the sucrose preference test upon inducing depression to rats, then, the sucrose preference test, alongside the open field and forced-swim tests at the end of the cohabitation period. Throughout the experiment, rats are given tags and are always returned to their cages after each test.
A few limitations to this method are the weak differences recorded between the experimental and control groups in the sucrose preference test and the irreversible traumatic outcome of the forced swim test. These may be worth considering for suitability before any future application of the protocol. Nonetheless, following the experiment, na&#239;ve rats developed contagion depression after 5 weeks of sharing the same cage with the CUS-exposed rats.

This protocol describes a new model by which healthy rats could contract depression over a given time periodthrough contagion by exposure to chronic unpredictable stressed (CUS) rats.

Um novo método para induzir um comportamento parecido com depressão em ratos

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is ...

Elevated Plus Maze for Mice

Although the mouse genome is now completely sequenced, the functions of most of the genes expressed in the brain are not known. The influence of a given gene on a specific behavior can be determined by behavioral analysis of mutant mice. If a target gene is expressed in the brain, behavioral phenotype of the mutant mice could elucidate the genetic mechanism of normal behaviors. The elevated plus maze test is one of the most widely used tests for measuring anxiety-like behavior. The test is based on the natural aversion of mice for open and elevated areas, as well as on their natural spontaneous exploratory behavior in novel environments. The apparatus consists of open arms and closed arms, crossed in the middle perpendicularly to each other, and a center area. Mice are given access to all of the arms and are allowed to move freely between them. The number of entries into the open arms and the time spent in the open arms are used as indices of open space-induced anxiety in mice. Unfortunately, the procedural differences that exist between laboratories make it difficult to duplicate and compare results among laboratories. Here, we present a detailed movie demonstrating our protocol for the elevated plus maze test. In our laboratory, we have assessed more than 90 strains of mutant mice using the protocol shown in the movie. These data will be disclosed as a part of a public database that we are now constructing.
Visualization of the protocol will promote better understanding of the details of the entire experimental procedure, allowing for standardization of the protocols used in different laboratories and comparisons of the behavioral phenotypes of various strains of mutant mice assessed using this test. 


The elevated plus maze test is one of the most widely used tests for measuring anxiety-like behavior in mice. Here, we present a movie showing the detailed procedures for conducting the test.

Labirinto elevado para Ratos

Although the mouse genome is now completely sequenced, the functions of most of the genes expressed in the brain are not known. The influence of a given ...

Biologia

Elevated Plus Maze Test Combined with Video Tracking Software to Investigate the Anxiolytic Effect of Exogenous Ketogenic Supplements

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The purpose of the method is to document the effect of various potential anxiolytic treatments on laboratory rodent models. The EPM test is based on the rodents&#39; proclivity toward protected, enclosed dark spaces and unconditioned fear of open spaces and heights, and their innate intense motivation to explore novel environments. The EPM test is a widely used behavioral test for investigating the anxiolytic or anxiogenic responses of rodents given drugs that are known to affect behavior. Observation demonstrating a decreased proportion of time spent on closed arms, an increased proportion of time spent on open arms, a reduced number of entries to closed arms, and an elevated number of entries to open arms measured by the EPM test may reflect reduced anxiety levels. Using this method, the effect of exogenous ketone supplements on anxiety-related behavior is tested in Sprague Dawley (SPD) rats. Exogenous ketone supplements are chronically fed to the rats for 83 days or subchronically and acutely orally gavaged, daily for 7 days, before conducting the EPM test. Behavioral data collection is performed using the SMART video tracking system by a blinded observer at the end of the treatments. The main findings indicate that the EPM test is an effective method to detect the ketone supplement-induced anxiolytic effect and can be considered a sensitive measure to assess changes in anxiety behavior associated with drug- or metabolic-based therapies.

Here, we present a protocol to investigate changes in the anxiety level of rodent animal models. The elevated plus maze (EPM) test, used together with a video tracking software, provides a reliable method to document the effect of various potential anxiolytic treatments in preclinical laboratory scenarios.

Elevado, mais teste combinado com vídeo que segue o Software para investigar o efeito ansiolítico dos suplementos cetogênica exógenos de labirinto

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The ...

Optogenetic Manipulation of Neuronal Activity to Modulate Behavior in Freely Moving Mice

Optogenetic modulation of neuronal circuits in freely moving mice affects acute and long-term behavior. This method is able to perform manipulations of single neurons and region-specific transmitter release, up to whole neuronal circuitries in the central nervous system, and allows the direct measurement of behavioral outcomes. Neurons express optogenetic tools via an injection of viral vectors carrying the DNA of choice, such as Channelrhodopsin2 (ChR2). Light is brought into specific brain regions via chronic optical implants that terminate directly above the target region. After two weeks of recovery and proper tool-expression, mice can be repeatedly used for behavioral tests with optogenetic stimulation of the neurons of interest.
Optogenetic modulation has a high temporal and spatial resolution that can be accomplished with high cell specificity, compared to the commonly used methods such as chemical or electrical stimulation. The light does not harm neuronal tissue and can therefore be used for long-term experiments as well as for multiple behavioral experiments in one mouse. The possibilities of optogenetic tools are nearly unlimited and enable the activation or silencing of whole neurons, or even the manipulation of a specific receptor type by light.
The results of such behavioral experiments with integrated optogenetic stimulation directly visualizes changes in behavior caused by the manipulation. The behavior of the same animal without light stimulation as a baseline is a good control for induced changes. This allows a detailed overview of neuronal types or neurotransmitter systems involved in specific behaviors, such as anxiety. The plasticity of neuronal networks can also be investigated in great detail through long-term stimulation or behavioral observations after optical stimulation. Optogenetics will help to enlighten neuronal signaling in several kinds of neurological diseases.

With optogenetic manipulation of specific neuronal populations or brain regions, behavior can be modified with high temporal and spatial resolution in freely moving animals. By using different optogenetic tools in combination with chronically implanted optical fibers, a variety of neuronal modulations and behavioral testing can be performed.

Manipulação optogenética da atividade neuronal para modular o comportamento em camundongos em movimento livre

Optogenetic modulation of neuronal circuits in freely moving mice affects acute and long-term behavior. This method is able to perform manipulations of ...

Validando um modelo murino da síndrome de Angelman usando testes comportamentais

Behavioral Characterization of an Angelman Syndrome Mouse Model

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of AS. We use the rotarod learning paradigm, detailed gait analysis, and nest building test to detect and characterize animal motor impairments. We test animal emotionality in the open field and elevated plus maze tests, as well as the affect in the tail suspension test. When AS mice are tested in the open field test, the results should be interpreted with care, since motor dysfunctions influence mouse behavior in the maze and alter activity scores.
The reproducibility and effectiveness of the presented behavioral tests has already been validated in several independent Uba3a mouse lines with different knockout variants, establishing this set of tests as an excellent validation tool in AS research. Models with the relevant construct and face validity will warrant further investigations to elucidate the pathophysiology of the disease and grant the development of causal treatments.

Autor em destaque: uma bateria de testes comportamentais altamente reproduzíveis para validar um modelo murino da síndrome de Angelman 

This manuscript presents a set of highly reproducible behavioral tests to validate an Angelman syndrome mouse model.

Caracterização comportamental de um modelo de camundongo com síndrome de Angelman

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of ...

Testes Animal ansiedade em ratos: efeitos de bordas de braço aberto e fechado do braço de parede transparência em elevados além de teste de labirinto

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O Becos Teste sucessiva de ansiedade em camundongos e ratos

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Diminui quantificar cognitivos causados pela radioterapia craniana