This work was supported by The National Institute of Health (NIH-2RO1NS033661) and by the Alabama Agricultural Experiment Station (HATCH ALA021-1-09017).

NOTA: Todos os procedimentos aqui realizados foram submetidos e aprovados pelo IACUC (Office of Compliance Research) e foram realizadas seguindo as diretrizes do NIH. Ratos usado no paradigma testes comportamentais eram ingênuas e não utilizados para outros testes. O C57BL / 6 selvagem Tipo e bater para fora ratos utilizados no presente protocolo têm sido descritos anteriormente 9 e os dados aqui apresentados são de que manuscrito. 1. Preparação da sala de teste e campo aberto Apparatus <ol><li> Usar um campo aberto labirinto unidade múltipla (OFM) que consiste em quatro câmaras de actividade foi usado para esta análise (Figura 1). Cada câmara medido 50 cm (comprimento) x 50 cm (largura) x 38 cm (altura) e foi feita a partir de alta densidade branco e plástico não-porosa. </li><li> Textura os andares do labirinto para a tração durante a deambulação, enquanto as paredes do labirinto eram lisas. Maze quadrantes foram completamente vazio para o objectivo do presente ensaio. Tendo em consideração o resto deste protocol, um único quadrante do labirinto acima descrito, irá ser utilizada para demonstrar a OFM. </li><li> Limpe a câmara com um 95% de etanol antes do uso e antes dos testes subsequentes para remover quaisquer indícios de cheiro deixados pelo mouse assunto anterior. </li><li> Permitir que o etanol se evaporar completamente antes de testar ratinhos. Isso pode levar 5-10 min entre cada sessão de testes. </li><li> Para esta análise, use o software SMART Video Rastreamento de Panlab / Harvard Apparatus para registrar e avaliar o movimento do mouse. NOTA: Qualquer câmera de monitoramento de vídeo comercial e software pode ser usado para rastrear o assunto de teste e avaliar os resultados do labirinto campo aberto. É muito importante para o usuário final para entender como calibrar e executar o software usado para cada análise individual. Independentemente da câmera de vídeo e software de rastreamento usado, os melhores resultados são obtidos quando a câmera e software estão corretamente calibrados de acordo com as instruções do fabricante. </li><li> Realize o teste ipadrão na iluminada sala capaz de conter o aparelho labirinto eo computador necessário para executar o software. Suspender a câmera de vídeo acima do labirinto ou anexando-o para o teto ou usando qualquer sistema de apoio elevada que permite que a lente da câmera para ver toda a área do labirinto (Figura 2). </li><li> Como administrador humano do teste, certifique-se de ter espaço suficiente no quarto para ser completamente não-observável pelos assuntos de teste no labirinto de modo a não influenciar o comportamento dos camundongos. </li></ol> 2. Preparar o Software para medir a atividade <ol><li> Abra o software de vídeo Rastreamento. </li><li> Uma vez que o software é aberto, mova o cursor até a opção &quot;rastreamento individual de cada disciplina&quot;, localizada na aba &quot;Aquisição de Dados&quot; e único clique para abrir esta opção. </li><li> Escolha a opção &quot;Antecedentes estático&quot;, localizado na parte inferior do ecrã. </li><li> Depois de escolher o &quot;fundo Static&quot; é escolhidon, que é necessário para usar o software para tomar uma imagem do labirinto antes da adição dos sujeitos de teste. Para fazer isso, mova o cursor para o botão &quot;Photo&quot; localizado na parte inferior da tela e único clique. NOTA: O software irá tirar uma foto do cenário sem o assunto de teste que será subtraído da imagem tomada durante o processo de acompanhamento. Isto resulta em apenas o movimento do sujeito a ser analisada pelo software. </li><li> Confirmar que a imagem de fundo tomada de cima é completamente removida pelo software de seguimento movendo o cursor para o botão de &quot;teste&quot;, localizado na parte inferior da tela e clicar uma vez. Um campo sólido branco será mostrado se a imagem de fundo é completamente removido a partir da imagem de rastreamento. Se as condições de iluminação ou mudar o labirinto é acidentalmente movido, você vai ver negros &quot;sombras&quot; neste domínio, indicando as duas imagens não se adequam perfeitamente coincidentes. Para remediar esta situação, basta repetir step 2.4 supra. </li><li> Depois de confirmar as configurações de fundo, use a opção Tempo &#39;para configurar o modo como o tempo é controlado durante a aquisição. Para fazer isso, mova o cursor para a aba &quot;Configuração&quot; e clique uma vez sobre o título &quot;intervalos&quot;. Use a nova janela aberta para inserir os parâmetros experimentais. </li><li> Para este protocolo, escolheu a opção &quot;tempo programado&quot; como período de acompanhamento de 10 min. Definir o &quot;período de latência&quot; a 5 segundos para permitir que o utilizador coloque o rato no centro do labirinto e afastar-se antes do início do acompanhamento. Introduzir o &quot;tempo de aquisição&quot; de 10 min durante a duração do teste. Defina o &quot;Pare de controle&quot; definida como &quot;Quando o tempo programado (10 min) é longo&quot;, que desliga-se automaticamente a câmera e a função de acompanhamento do software. </li><li> Mova o cursor para o botão &quot;Fechar&quot; depois de todos os tempos foram definidos para fechar a janela. Agora você está pronto para iniciar o testeing procedimento. </li></ol> 3. Administração do Aberto Campo de Provas NOTA: O pacote de software usado neste protocolo permite o rastreamento de até 16 ratos individuais ao mesmo tempo. Para facilidade de realização e, como mencionado acima, o protocolo discutidos aqui é para um único rato usando um único quadrante da OFM. Para o equipamento em uso para este protocolo, um máximo de 4 ratos individuais poderiam ser rastreados usando cada quadrante do labirinto. Se utilizando um labirinto multi-gabinete, depois de colocar o primeiro mouse assunto no quadrante definido, coloque os ratos restantes em seu respectivo quadrante labirinto para análise de rastreamento. Para efeitos do presente protocolo, mais instrução será específica para um único quadrante do labirinto. <ol><li> Traga os ratos em suas gaiolas de sua sala que abriga a sala de ensaios. Permitir que a ratinhos para se aclimatarem ao ambiente do processo durante um período mínimo de 30 minutos antes de se iniciar o teste. </li><li> Retirar um único mOuse desde a gaiola, segurando gentilmente sua cauda e coloque o mouse no meio do labirinto campo aberto ao mesmo tempo em ativar o software SMART simplesmente clicando no botão Iniciar para começar a acompanhar o movimento do mouse. É normal que o mouse para mover imediatamente para as paredes periféricas do labirinto e do calendário de liberação e captura do mouse rastreamento deverá coincidir para gravar este movimento. </li><li> Permitir livre e ininterrupto movimento do mouse assunto durante o respectivo quadrante do labirinto para um único período de 10 minutos durante os quais, o software de rastreamento irá gravar o movimento (Figura 3). </li><li> No final do período de teste, pegar o mouse assunto suavemente, removendo-o do labirinto e devolvê-lo à sua gaiola. </li><li> Antes da limpeza do labirinto, contar visualmente os peletes boli fecais presentes no labirinto e gravar os números manualmente para análise posterior. </li><li> Remova todas as pelotas fecais e limpe todas as manchas de urina. Pulverizar o chão e as paredes do quadrante labirinto com 95% de etanol e limpe com uma toalha de papel. Permitir que a solução de etanol para secar completamente antes de testar outros ratinhos. </li><li> Repita o procedimento com o próximo mouse. </li></ol> 4. Medição e Análise do Comportamento Durante procedimentos de teste NOTA: Para a medição, três aspectos do comportamento campo aberto são facilmente caracterizados utilizando este protocolo (ver discussão). Uma breve instrução sobre como acessar essas medições no software de monitoramento de vídeo segue. <ol><li> Na tela principal do software SMART, mova o cursor para a &quot;Zones&quot; tab e único clique &quot;Definition&quot; para abrir o Editor Zone. </li><li> Siga as instruções detalhadas no Manual do Usuário do software SMART que definir as zonas ou grades para sobrepor sobre os caminhos de rastreamento. Aqui, o software foi utilizado para definir uma grade 5 x 5 de 10 centímetros quadrados cobrindo o chão do labirinto ( <strong&gt; Figura 4). Certifique-se de salvar o arquivo de zona criado antes de fechar o Editor Zone. </li><li> Na tela principal do software SMART, mova o cursor para a aba &quot;Análise&quot; e único clique para abrir a janela de Análise de Dados. </li><li> Mova o cursor até a guia &quot;Arquivo&quot; e abra o arquivo Zona criado acima. </li><li> Mova o cursor para a aba &quot;Configuração&quot; e abra a opção &quot;Track Analysis&quot;. Isso abrirá a janela &quot;configuração análise Single-assunto&quot;. <ol><li> Mova o cursor para a aba &quot;Standard&quot; e mova o parâmetro &quot;Distância Percorrida&quot; na caixa de parâmetros disponíveis, (lado esquerdo) para a caixa de parâmetros incluídos (lado direito). </li><li> Mova o cursor para a aba &quot;zonas Transitions&quot; e mover todos os parâmetros apropriados para a caixa de parâmetros incluídos como acima. </li><li> Verifique se a caixa &quot;Track Full&quot; é verificada na parte inferior da janela. </li><li> Move o cursor para o botão OK e feche a janela de configuração de análise individual de cada disciplina. NOTA: Dependendo da análise que deseja realizar, muitas outras opções podem ser escolhidos nesta janela de explorar dados de análise. Leia o manual do seu programa específico do Usuário detalhado de parâmetros para determinar o que são os mais importantes para a análise de seus dados. </li></ol></li><li> Sob a guia Arquivo no canto superior esquerdo da janela do programa, abra a janela pista sujeito único e coloque uma marca de seleção ao lado de todas as faixas que está sendo analisado. Mova o cursor para o botão Check Mark, na parte superior da janela e clique para fechar a janela de Pista Explorer. </li><li> Mova o cursor para o botão &quot;Go&quot; na janela de Análise de Dados e único clique para iniciar a análise dos dados da faixa. </li><li> Análise de dados pode ser a saída tanto como arquivos de texto ASCII, ou pode ser exportado diretamente em uma planilha do Excel. Use as ferramentas de saída do programa de software que você está usando para a saída do data para seu próprio uso. NOTA: A distância total percorrida e do tempo passado nas zonas indicadas será a saída seguindo as etapas de análise de dados descritos acima. Mais uma vez, salienta-se que os passos para chegar a estas medições representados aqui serão diferentes dependendo do software do usuário utilizado. Mas os dados em si e à interpretação dos resultados deve ser independente semelhante do programa de software usado. Também é importante notar que o viés administrador do teste é removido este protocolo como todos os dados coletados é quantificado dados medidos pelo software e não o administrador. Assim, não há nenhum elemento qualificável com os dados coletados, como descrito. </li></ol>

<ol>
	<li>Hall, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotional+behavior+in+the+rat:+defecation+and+urination+as+measures+of+individual+differences+in+emotionality.">Emotional behavior in the rat: defecation and urination as measures of individual differences in emotionality.</a> J. Comp. Psychol. 18, 385-403 (1934).</li><li>Walsh, R. N., Cummins, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+test:+a+critical+review.">The open field test: a critical review.</a> Psychol. Bull. 83, 482-504 (1976).</li><li>Prut, L., Belzung, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+as+a+paradigm+to+measure+the+effects+of+drugs+on+anxiety-like+behaviors:+a+review.">The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review.</a> Eur. J. of Pharm. 463, 3-33 (2003).</li><li>Choleris, E., Thomas, A. W., Kavaliers, M., Prato, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+detailed+ethological+analysis+of+the+mouse+open+field+test:+effects+of+diazepam,+chlordiazpoxide+and+an+extremely+low+frequency+pulsed+magnetic+field.">A detailed ethological analysis of the mouse open field test: effects of diazepam, chlordiazpoxide and an extremely low frequency pulsed magnetic field.</a> Neurosci. Biobehav. Rev. 25, 235-260 (2001).</li><li>Ramos, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+of+anxiety:+do+I+need+multiple+tests.">Animal Models of anxiety: do I need multiple tests.</a> TIPS. 29, 493-498 (2008).</li><li>Archer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+for+emotionality+in+rats+and+mice:+a+review.">Tests for emotionality in rats and mice: a review.</a> Anim. Behav. 21, 205-235 (1973).</li><li>Gray, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotionality+in+male+and+female+rodents:+a+reply+to+Archer.">Emotionality in male and female rodents: a reply to Archer.</a> Brit. J. Psych. 70, 425-440 (1979).</li><li>Carola, V., D&#8217;Olimpio, F., Brunamonti, E., Mangia, F., Renzi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+elevated+plus-maze+and+open-field+tests+for+the+assessment+of+anxiety-related+behavior+in+inbred+mice.">Evaluation of the elevated plus-maze and open-field tests for the assessment of anxiety-related behavior in inbred mice.</a> Behav. Brain Res. 134, 49-57 (2002).</li><li>Ramesh Babu, J., Seibenhener, M. L., Peng, J., Strom, A. L., Kemppainen, R., Cox, N., Zhu, H., Wooten, M. C., Diaz-Meco, M. T., Moscat, J., Wooten, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+inactivation+of+p62+leads+to+accumulation+of+hyperphosphorylated+tau+and+neurodegeneration.">Genetic inactivation of p62 leads to accumulation of hyperphosphorylated tau and neurodegeneration.</a> J. Neurochem. 106, 107-120 (2008).</li><li>Simon, P., Dupuis, R., Costentin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmotaxis+as+an+index+of+anxiety+in+mice:+influence+of+dopaminergic+transmissions.">Thigmotaxis as an index of anxiety in mice: influence of dopaminergic transmissions.</a> Behav. Brain Res. 61, 59-64 (1994).</li><li>Denenberg, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-field+behavior+in+the+rat:+what+does+it+mean.">Open-field behavior in the rat: what does it mean.</a> Ann. N.Y. Acad. Sci. 159, 852-859 (1969).</li><li>Stanford, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+test:+reinventing+the+wheel.">The open field test: reinventing the wheel.</a> J. Psychopharm. 21, 134-135 (2007).</li><li>Ennaceur, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+of+unconditional+anxiety+&#8211;+pitfalls+and+disappointments.">Tests of unconditional anxiety &#8211; pitfalls and disappointments.</a> J. Phys. Behav. 135, 55-71 (2014).</li><li>Borta, A., Schwarting, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitory+avoidance,+pain+reactivity,+and+plus-maze+behavior+in+Wistar+rats+with+high+versus+low+rearing+activity.">Inhibitory avoidance, pain reactivity, and plus-maze behavior in Wistar rats with high versus low rearing activity.</a> J. Phys. Behav. 84, 387-396 (2005).</li><li>Costall, B., Jones, B. J., Kelly, M. E., Naylor, R. J., Tomkins, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploration+of+mice+in+a+black+and+white+test+box:+validation+as+a+model+of+anxiety.">Exploration of mice in a black and white test box: validation as a model of anxiety.</a> Pharmacol. Biochem. Behav. 32, 777-785 (1989).</li><li>Bouwknecht, J. A., Paylor, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pitfalls+in+the+interpretation+of+genetic+and+pharmacological+effects+on+anxiety-like+behavior+in+rodents.">Pitfalls in the interpretation of genetic and pharmacological effects on anxiety-like behavior in rodents.</a> Behav. Pharm. 19, 385-402 (2008).</li><li>Webster, D. G., Baumgardner, D. J., Dewsbury, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open+field+behavior+in+eight+taxa+of+muriod+rodents.">Open field behavior in eight taxa of muriod rodents.</a> Bull. Psychonom. Soc. 13, 90-92 (1979).</li><li>Wilson, R. C., Vacek, T., Lanier, D. L., Dewsbury, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open+field+behavior+in+muroid+rodents.">Open field behavior in muroid rodents.</a> Behav. Biol. 17, 495-506 (1976).</li><li>Crawley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+phenotyping+of+transgenic+and+knockout+mice:+experimental+designs+and+evaluation+of+general+health,+sensory+functions,+motor+abilities+and+specific+behavioral+tests.">Behavioral phenotyping of transgenic and knockout mice: experimental designs and evaluation of general health, sensory functions, motor abilities and specific behavioral tests.</a> Brain Res. 835, 18-26 (1999).</li><li>Simon, P., Dupuis, R., Costentin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmotaxis+as+an+index+of+anxiety+in+mice.+influence+of+dopaminergic+transmissions.">Thigmotaxis as an index of anxiety in mice. influence of dopaminergic transmissions.</a> Behav. Brain Res. 61, 59-64 (1994).</li><li>Miller, B. H., Schultz, L. E., Gulati, A., Su, A. I., Pletcher, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+characterization+of+a+genetically+diverse+panel+of+mice+for+behavioral+despair+and+anxiety.">Phenotypic characterization of a genetically diverse panel of mice for behavioral despair and anxiety.</a> PLoS One. 5, e14458 (2010).</li><li>Tannenbaum, B., Anisman, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+chronic+intermittent+challenges+in+stressor-susceptible+and+resilient+strains+of+mice.">Impact of chronic intermittent challenges in stressor-susceptible and resilient strains of mice.</a> Biol. Psych. 53, 292-303 (2003).</li><li>Lister, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethologically-based+animal+models+of+anxiety+disorders.">Ethologically-based animal models of anxiety disorders.</a> Pharmacol. Ther. 46, 321-340 (1990).</li><li>Kulesskaya, N., Voikar, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+mouse+anxiety-like+behavior+in+the+light-dark+box+and+open-field+arena:+role+of+equipment+and+procedure.">Assessment of mouse anxiety-like behavior in the light-dark box and open-field arena: role of equipment and procedure.</a> Phys. Behav. 133, 30-38 (2014).</li><li>Han, H., Du, W., Zhou, B., Zhang, W., Xu, G., Niu, R., Sun, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+chronic+fluoride+exposure+on+object+recognition+memory+and+mRNA+expression+of+SNARE+complex+in+hippocampus+of+male+mice.">Effects of chronic fluoride exposure on object recognition memory and mRNA expression of SNARE complex in hippocampus of male mice.</a> Biol. Trace Elem. Res. 158, 58-64 (2014).</li><li>Barrow, P., Leconte, I. <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+body+weight+on+open+field+and+swimming+maze+performance+during+the+post-weaning+period+in+the+rat.">The influence of body weight on open field and swimming maze performance during the post-weaning period in the rat.</a> Lab. Animals. 30, 22-27 (1996).</li></ol>

The authors have nothing to disclose.

O campo aberto Maze é uma das plataformas mais utilizadas em estudos de comportamento dos animais. Uma série de importantes parâmetros convencionais e etológicas 2,4 podem ser coletadas e analisadas durante a execução da OFM. Estes dados permitem que o pesquisador para medir comportamentos que variam de actividade global locomotor a comportamentos emocionais relacionados à ansiedade 8. No entanto, o uso de OFM não é sem suas deficiências. Uma questão de confusão é a ampla gama de vari?...

The Open Field Maze (OFM) was initially developed in 1934 as a test to measure emotionality in rodents1. It has attained the status of being one of the most widely used measures of behavior in animal psychology2. It provides an easy and fairly rapid assessment of well-defined behaviors requiring no training to the test subject and little to no specialized training for the human administering the test. These attributes have led to wide-spread use of the open field maze in research extended to other animal species such as calves, pigs, rabbits, primates, honeybees and lobsters3. Part of its popularity arises from the fact that the psychological and physiological concepts underlying the tests are generally straight-forward and well understood. For example, it has been postulated that evolutionary forces have selected for a common response in animals such that most species display anxiety-mediated fear or flight responses to specific stimuli. Rodents for example, show distinct aversions to large, brightly lit, open and unknown environments4. We can assume they have been phylogenetically conditioned to see these types of environments as dangerous. All of these features are incorporated in the open field maze and form the basis of its use in behavioral paradigm testing.
An open field maze consists of a wall-enclosed area that is of sufficient height to prevent the subject from escaping. Typical maze shapes are circular or square with an area large enough, based on the size of the subject tested, to elicit a feeling of openness in the center of the maze. A number of variables can be scored in the open field maze with most parameters involving differing types of motor activity2. Ambulation is the most common behavior studied but others such as latency or rearing can also be measured. Most often, rodent behavior is analyzed in a bare maze. However, the addition of objects, either one or many to the maze floor, adds the ability to see how the subject interacts with novel additional stimuli2. Relevant parameters when objects are presented are typically the number of approaches to an object or in some cases, preference or aversion for one object over another.
Many behavioral tests of anxiety are based on the subject animal&#8217;s body activity and locomotion5. Interpreting behavioral tests for emotionality while separating non-emotional confounding factors, such as motor activity, has been the subject of intense debate6,7. As the OFM was originally described, two measures of emotionality can be deduced, locomotor activity and fecal boli deposits or defecation1. However, these two measures have been shown in some studies to be unrelated supporting the conclusion that emotionality in rodents is multidimensional5. Regardless, discrepancies in the literature regarding these measures and emotionality or anxiety in mouse models may be attributed to differences in analysis criteria or differences in testing procedures. Studies have conclusively linked results from OFM analysis with other measures of anxiety when comparing mouse models8.

<table border="0" cellpadding="0" cellspacing="0" width="631">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:160px;">Multi Unit Open Field Test</td>
			<td style="width:171px;">San Diego Instruments, Inc.</td>
			<td style="width:109px;">White 7001-0354</td>
			<td style="width:192px;">Any single or multi unit open field maze can be used</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:160px;">SMART DT Tracking Software</td>
			<td style="width:171px;">PanLab/Harvard Apparatus</td>
			<td style="width:109px;">76-0695</td>
			<td style="width:192px;">Any tracking software can be utilized with this protocol</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:160px;">Sony 990x Video Camera Recorder</td>
			<td style="width:171px;">Sony</td>
			<td style="width:109px;">CCD-TRV328</td>
			<td style="width:192px;">Any suitable video camera can be attached to computer for recording tracking profiles.</td>
		</tr>
	</tbody>
</table>

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.

O número médio de indivíduos por estirpe de ratinhos testados, na maioria dos casos é de aproximadamente 20 para gerar relevância estatística suficiente. No entanto, este número pode estar na gama de 8-30, dependendo da disponibilidade do rato. Dependendo da medida ou comparações necessário, também é favorável utilizar pareados por idade. O primeiro e talvez o mais importante parâmetro específico para medir no campo aberto Maze é a distância ambulatorial total. Enquanto a un...

Watch this Scientific Journal Video about Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice at JoVE.com

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

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 ...

use-open-field-maze-to-measure-locomotor-anxiety-like-behavior

Research

JoVE Journal

Behavior

72.0K visualizações.  Auburn University. A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

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

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to the mouse becoming increasingly important. But researchers found that some strains of mutant mice were prone to problems like passively floating or diving when they were tested in the Morris water maze11. This was unsurprising considering their natural habitat; rats swim naturally (classically, the "sewer rat"), whereas mice evolved in the dry areas of central Asia.
To overcome these problems, it was considered whether shallow water would be a sufficient stimulus to provide escape motivation for mice. This would also avoid the problems of drying the small creatures with a towel and then putting them in a heated recovery chamber to avoid hypothermia, which is a much more serious problem than with rats; the large ratio of surface area to volume of a mouse makes it particularly vulnerable to rapid heat loss.
Another consideration was whether a more natural escape strategy could be used, to facilitate learning. Since animals that fall into water and swim away from the safety of the shore are unlikely to pass on their genes, animals have evolved a natural tendency to swim to the edge of a body of water. The Morris water maze, however, requires them to swim to a hidden platform towards the center of the maze - exactly opposite to their evolved behavior. Therefore the paddling maze should incorporate escape to the edge of the apparatus. This feature, coupled with the use of relatively non-aversive shallow water, embodies the "Refinement" aspect of the "3 Rs" of Russell and Burch8.
Various types of maze design were tried; the common feature was that the water was always shallow (2 cm deep) and escape was via a tube piercing the transparent wall of the apparatus. Other tubes ("false exits") were also placed around the walls but these were blocked off. From the inside of the maze all false exits and the single true exit looked the same. Currently a dodecagonal (12-sided) maze is in use in Oxford, with 12 true/false exits set in the corners. In a recent development a transparent paddling Y-maze has been tested successfully.

Shallow Water Paddling Variants of Water Maze Tests in Mice

Mice can swim, but many strains appear to find this activity stressful. To overcome this problem mazes have been devised where escape from shallow water is used to motivate behaviour. These have been demonstrated to support learning at least as good as the traditional and widely used Morris water maze.

Água (Palmatória) Variantes de água rasa testes de labirinto em ratos

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to ...

Comportamento

Use of the Operant Orofacial Pain Assessment Device (OPAD) to Measure Changes in Nociceptive Behavior

We present an operant system for the detection of pain in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) assesses pain behaviors in a more clinically relevant way by not relying on reflex-based measures of nociception. Food fasted, hairless (or shaved) rodents are placed into a Plexiglas chamber which has two Peltier-based thermodes that can be programmed to any temperature between 7 &deg;C and 60 &deg;C. The rodent is trained to make contact with these in order to access a reward bottle. During a session, a number of behavioral pain outcomes are automatically recorded and saved. These measures include the number of reward bottle activations (licks) and facial contact stimuli (face contacts), but custom measures like the lick/face ratio (total number of licks per session/total number of contacts) can also be created. The stimulus temperature can be set to a single temperature or multiple temperatures within a session. The OPAD is a high-throughput, easy to use operant assay which will lead to better translation of pain research in the future as it includes cortical input instead of relying on spinal reflex-based nociceptive assays.

Use of the Operant Orofacial Pain Assessment Device OPAD to Measure Changes in Nociceptive Behavior

We present a user-friendly, high-throughput operant system for the evaluation of pain behaviors in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) can assess pain through a reward/conflict paradigm thus providing a more humane way of testing. This protocol will yield more clinically relevant and translational data from rodents.

O uso do dispositivo de Avaliação de Dor Orofacial operante (OPAD) para medir as mudanças no comportamento nociceptivo

We present an operant  system for the detection of pain in awake, conscious rodents. The Orofacial  Pain Assessment Device (OPAD) assesses pain behaviors ...

Use of an Eight-arm Radial Water Maze to Assess Working and Reference Memory Following Neonatal Brain Injury

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food deprivation, and may introduce scent cue confounds. The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs. Specifically, subjects are required to avoid the arms previously used for escape during each testing day (working memory) as well as avoid the fixed arms, which never contain escape platforms (reference memory). Re-entries into arms that have already been used for escape during a testing session (and thus the escape platform has been removed) and re-entries into reference memory arms are indicative of working memory deficits. Alternatively, first entries into reference memory arms are indicative of reference memory deficits. We used this maze to compare performance of rats with neonatal brain injury and sham controls following induction of hypoxia-ischemia and show significant deficits in both working and reference memory after eleven days of testing. This protocol could be easily modified to examine many other models of learning impairment.

The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs.

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food ...

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing locomotive impairment in animal models of neuromuscular disease and efficacy of therapeutic drugs that may improve locomotion and/or muscle function. The open field activity measurement provides a different measure than muscle strength, which is commonly assessed by grip strength measurements. It can also show how drugs may affect other body systems as well when used with additional outcome measures. In addition, measures such as total distance traveled mirror the 6 min walk test, a clinical trial outcome measure. However, open field activity monitoring is also associated with significant challenges: Open field activity measurements vary according to animal strain, age, sex, and circadian rhythm. In addition, room temperature, humidity, lighting, noise, and even odor can affect assessment outcomes. Overall, this manuscript provides a well-tested and standardized open field activity SOP for preclinical trials in animal models of neuromuscular diseases. We provide a discussion of important considerations, typical results, data analysis, and detail the strengths and weaknesses of open field testing. In addition, we provide recommendations for optimal study design when using open field activity in a preclinical trial.

Open field activity levels are used to assess locomotive and behavioral activity levels. This protocol provides a well-designed, standardized protocol to use in preclinical trials for neuromuscular disorders.

Medidas comportamentais e do aparelho locomotor Usando um sistema aberto Activity Monitoring Campo de Doenças músculo-esquelético

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

The Attentional Set Shifting Task: A Measure of Cognitive Flexibility in Mice

Cognitive impairment, particularly involving dysfunction of circuitry within the prefrontal cortex (PFC), represents a core feature of many neuropsychiatric and neurodevelopmental disorders, including depression, post-traumatic stress disorder, schizophrenia and autism spectrum disorder. Deficits in cognitive function also represent the most difficult symptom domain&#160;to successfully treat, as serotonin reuptake inhibitors and tricyclic antidepressants have only modest effects. Functional neuroimaging studies and postmortem analysis of human brain tissue implicate the PFC as being a primary region of dysregulation in patients with these disorders. However, preclinical behavioral assays used to assess these deficits in mouse models which can be readily manipulated genetically and could provide the basis for studies of new treatment avenues have been underutilized. Here we describe the adaptation of a behavioral assay, the attentional set shifting task (AST), to be performed in mice to assess prefrontal cortex mediated cognitive deficits. The neural circuits underlying behavior during the AST are highly conserved across humans, nonhuman primates and rodents, providing excellent face, construct and predictive validity.

The goal of this protocol is to perform a behavioral assay such as the attentional set shifting task (AST) to assess prefrontal cortex-mediated cognitive flexibility in mice.

A Task Attentional Set Shifting: uma certa flexibilidade cognitiva em ratos

Cognitive impairment, particularly involving dysfunction of circuitry within the prefrontal cortex (PFC), represents a core feature of many ...

Place and Response Learning in the Open-field Tower Maze

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for examining how rats learn to use a place- or response-learning to successfully navigate in an open-field arena. Additionally, this protocol describes how the OFTM differs from other behavioral maze paradigms that are commonly used to study spatial learning in rodents. The OFTM described in this article was adapted from the one previously described by Cole, Clipperton, and Walt (2007). Specifically, the OFTM was created to test spatial learning in rodents without the experimenter having to consider how &#8220;stress&#8221; might play a role as a confounding variable. Experiments have shown that stress-alone can significantly affect cognitive function1. The representative results section contains data from an experiment that used the OFTM to examine the effects of estradiol treatment on place- and response-learning in adult female Sprague Dawley rats2. Future studies will be designed to examine the role of the hippocampus and striatum in place- and response-learning in the OFTM.

The Open-field Tower Maze (OFTM) is a maze created to study the behavioral and neural mechanisms of spatial learning (e.g., place- or response-learning) in rats. This maze is especially useful for experimenters who want to use a non-stressful maze paradigm to investigate spatial learning in their research.

Coloque e Aprendizagem Response no Open-campo Torre Maze

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for ...

Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a series of basic techniques are described to enable observations of the reproductive behavior of a wild-caught gobiid fish, as a model, kept in an aquarium. The method focuses on three steps: collection, transport, and observations of reproductive ecology of a substrate spawner. Essential aspects of live fish collection and transport are (1) preventing injury to the fish, and (2) careful acclimation to the aquarium. Preventing harm through injuries such as scratches or a sudden change of water pressure is imperative when collecting live fish, as any physical damage is likely to negatively affect the survival and later behavior of the fish. Careful acclimation to aquaria decreases the incidence death and mitigates the shock of transport. Observations during captive rearing include (1) the identification of individual fish and (2) monitoring spawned eggs without negative effects to the fish or eggs, thereby enabling detailed investigation of the study species' reproductive ecology. The subcutaneous injection of a visible implant elastomer (VIE) tag is a precise method for the subsequent identification of individual fish, and it can be used with a wide size range of fish, with minimal influence on their survival and behavior. If the study species is a substrate spawner that deposits adhesive eggs, an artificial nest site constructed from polyvinyl chloride (PVC) pipe with the addition of a removable waterproof sheet will facilitate counting and monitoring the eggs, lessening the investigator's influence on the nest-holding and egg-guarding behavior of the fish. Although this basic method entails techniques that are seldom mentioned in detail in research articles, they are fundamental for undertaking experiments that require the captive rearing of a wild fish.

A series of basic methods to enable the study of the reproductive ecology of fish kept in aquaria are described. These are useful protocols for collecting fish using SCUBA, transporting live fish, and observing the reproductive behavior of wild-caught fish kept in aquaria.

Métodos Básicos para o Estudo da Ecologia Reprodutiva do Peixe em Aquário

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a ...

Central and Divided Visual Field Presentation of Emotional Images to Measure Hemispheric Differences in Motivated Attention

Two dominant theories on lateralized processing of emotional information exist in the literature. One theory posits that unpleasant emotions are processed by right frontal regions, while pleasant emotions are processed by left frontal regions. The other theory posits that the right hemisphere is more specialized for the processing of emotional information overall, particularly in posterior regions.
Assessing the different roles of the cerebral hemispheres in processing emotional information can be difficult without the use of neuroimaging methodologies, which are not accessible or affordable to all scientists. Divided visual field presentation of stimuli can allow for the investigation of lateralized processing of information without the use of neuroimaging technology. 
This study compared central versus divided visual field presentations of emotional images to assess differences in motivated attention between the two hemispheres. The late positive potential (LPP) was recorded using electroencephalography (EEG) and event-related potentials (ERPs) methodologies to assess motivated attention. Future work will pair this paradigm with a more active behavioral task to explore the behavioral impacts on the attentional differences found.

This study compared central versus divided visual field presentations of emotional images to assess differences in motivated attention between the two hemispheres. The late positive potential (LPP) was recorded using electroencephalography (EEG) and event-related potentials (ERPs) methodologies to assess motivated attention.

Central e dividida campo Visual apresentação de imagens emocionais para medir diferenças hemisféricas em atenção motivada

Two dominant theories on lateralized processing of emotional information exist in the literature. One theory posits that unpleasant emotions are processed ...

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

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 science, conflicting outcomes are often provided from different laboratories. To identify reasons for the different outcomes, we previously focused on arm features, which differ between laboratories, most notably the presence/absence of ledges at the sides of open arms and the transparency/opaqueness of closed arm walls. In a previous report, we used a custom designed container to compare rat behavior on different combinations of open and closed arm designs under otherwise identical experimental conditions, and showed that differences in arm features interfere with experimental outcomes. In brief, open arm ledges significantly increased anxiety-like behavior in rats, while transparent arms may also have decreased this behavior. Furthermore, we verified a higher detection sensitivity of the effect of an anxiolytic drug in a combination of no-ledged open arm + opaque walled closed arm compared with a combination of ledged open arm + transparent walled closed arm. In this report, we introduce our protocol for the elevated plus maze test, together with discussion of the key results from the previous report and our experimental experience. We believe this report will provide useful information for researchers who have employed or who plan to use the elevated plus maze in their studies.

Rats' anxiety-like behavior tested by the elevated plus maze varies highly depending on the testing environment. Here we show how apparatus settings, especially arm structure, affect behavior on the maze, and we explain a reliable protocol to detect effects of anxiolytic drugs. This report will contribute to anxiety research.

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

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 ...

Analyzing Spatial Learning and Prosocial Behavior in Mice Using the Barnes Maze and Damsel-in-Distress Paradigms

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. The Barnes maze can also assess other mouse behaviors, such as general motivation to escape from the maze platform and exploratory behavior. The Barnes maze can measure whether a genetic mutation or environmental variable can impact the acquisition and retention of spatial memories, as well as provide information about the search strategy employed by the mice. Here we use the Barnes maze to detect a memory deficit in adult mice following a single developmental ethanol exposure event. The newly described Damsel-in-Distress paradigm exposes a male mouse to a female mouse trapped in a chamber in the open center field of the arena. It provides an opportunity for the mouse to socially respond to the trapped female and exhibit prosocial behavior. The Damsel-in-Distress paradigm can also be used to examine mouse behavior in a novel arena and measure locomotor activity. Both the Barnes Maze and the Damsel-in-Distress protocols require minimal financial investment and most aspects of the tests can be constructed from common lab supplies. These flexible and accessible tools can also be used to detect behavioral changes over the course of development.

This protocol measures spatial learning and memory using the Barnes maze. A novel Damsel-in-Distress paradigm is used to assess locomotor activity and prosocial behavior in mice.

Analisando a aprendizagem espacial e comportamento pró-social nos ratos usando o labirinto de Barnes e paradigmas de donzela em apuros

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. ...