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 los procedimientos realizados aquí fueron presentados y aprobados por IACUC (Oficina de Cumplimiento de Investigación) y se llevaron a cabo siguiendo las directrices del NIH. Los ratones utilizados en el paradigma de las pruebas de comportamiento eran ingenuos y no se utiliza para otras pruebas. La C57BL / 6 de tipo salvaje y ratones knock out utilizado en este protocolo se han descrito anteriormente 9 y los datos presentados aquí son de ese manuscrito. 1. Preparación de la sala de pruebas y Aparato campo abierto <ol><li> Utilice un laberinto campo abierto unidad múltiple (OFM) que consta de cuatro cámaras de actividad se utilizó para este análisis (Figura 1). Cada cámara de medida 50 cm (longitud) x 50 cm (ancho) x 38 cm (altura) y se hizo desde el blanco de alta densidad y de plástico no poroso. </li><li> Textura Las plantas del laberinto para la tracción durante la deambulación mientras que las paredes del laberinto eran lisas. Cuadrantes Maze eran completamente vacía para el propósito de esta prueba. En consideración a el resto de este protocol, un único cuadrante del laberinto se ha descrito anteriormente se utilizará para demostrar la OFM. </li><li> Limpiar la cámara con un 95% de etanol antes de su uso y antes de las pruebas posteriores para eliminar cualquier olor pistas dejadas por el sujeto ratón anterior. </li><li> Deje que el etanol se evapore completamente antes de probar los ratones. Esto puede tomar 5-10 minutos entre cada sesión de evaluación. </li><li> Para este análisis, utilice el software SMART Seguimiento por vídeo de Aparato PanLab / Harvard para registrar y evaluar el movimiento del ratón. NOTA: Cualquier cámara de seguimiento de vídeo comercial y el software se pueden utilizar para realizar un seguimiento del sujeto de prueba y evaluar los resultados del laberinto campo abierto. Es muy importante para el usuario final para comprender cómo calibrar y ejecutar el software utilizado para cada análisis individual. Independientemente de la cámara de vídeo y software de seguimiento utilizado, se obtienen mejores resultados cuando la cámara y el software están calibrados correctamente de acuerdo con las instrucciones del fabricante. </li><li> Realice la prueba inorma na sala capaz de contener el aparato laberinto y el equipo necesario para ejecutar el software iluminado. Suspender la cámara de vídeo por encima del laberinto bien asociándolo al techo o utilizando cualquier sistema de soporte elevada que permite el lente de la cámara para ver toda la zona de laberinto (Figura 2). </li><li> Como administrador humano de la prueba, asegúrese de tener suficiente espacio en la sala de estar completamente inobservable por los sujetos de prueba en el laberinto para no influir en el comportamiento de los ratones. </li></ol> 2. Preparar el software para medir la actividad <ol><li> Abra el software de seguimiento de vídeo. </li><li> Una vez que el software se abre, mover el cursor a la opción &quot;seguimiento de un solo sujeto&quot; que se encuentra bajo la pestaña de &quot;Adquisición de datos&quot; y solo clic para abrir esta opción. </li><li> Elija la opción &quot;fondo estático&quot;, ubicado en la parte inferior de la pantalla. </li><li> Después de elegir &quot;fondo estático&quot; es elegidon, es necesario utilizar el software para tomar una imagen del laberinto antes de la adición de los sujetos de prueba. Para ello, mueva el cursor en el botón &quot;Foto&quot;, ubicado en la parte inferior de la pantalla y solo clic. NOTA: El software tomará una foto del escenario sin el sujeto de prueba que se restará de la imagen tomada durante el proceso de seguimiento. Esto resulta en sólo el movimiento del sujeto que se analiza por el software. </li><li> Confirme que la imagen de fondo tomada anteriormente se extrae completamente el software de seguimiento al mover el cursor en el botón &quot;Test&quot; situado en la parte inferior de la pantalla y haciendo clic una vez. Un campo sólido blanco se mostrará si la imagen de fondo se elimina completamente de la imagen de seguimiento. Si las condiciones de iluminación cambian o el laberinto es accidentalmente movido, podrás ver &quot;sombras&quot; negros en este campo que indica las dos imágenes no coinciden perfectamente. Para remediar esta situación, simplemente repita step 2.4 anteriormente. </li><li> Después de confirmar la configuración del fondo, utilice la opción de sincronización para configurar el modo de tiempo se controla durante la adquisición. Para ello, mueva el cursor a la pestaña &quot;Configuración&quot; y haga clic una vez en el título &quot;Tiempos&quot;. Utilice la nueva ventana abierta para entrar en parámetros experimentales. </li><li> Para este protocolo, eligió la opción &quot;Tiempo programado&quot; como el período de seguimiento de 10 min. Ajuste el &quot;período de latencia&quot; a 5 segundos para dar tiempo al usuario colocar el ratón en el centro del laberinto y alejarse antes de iniciar el seguimiento. Introduzca el &quot;Tiempo de adquisición&quot; de 10 min para la duración de la prueba. Ajuste la &quot;Llave de Paso&quot; en &quot;Cuando el tiempo programado (10 min) ha terminado&quot;, que se apagará automáticamente la cámara y la función de seguimiento del software. </li><li> Mueva el cursor sobre el botón &quot;Cerrar&quot; después de todos los tiempos se han establecido para cerrar la ventana. Ahora está listo para iniciar la pruebaing procedimiento. </li></ol> 3. Administración de la prueba de campo abierto NOTA: El paquete de software utilizado en este protocolo permite el seguimiento de hasta 16 ratones individuales al mismo tiempo. Para facilidad de realización y como se mencionó anteriormente, el protocolo descrito aquí es para un solo ratón utilizando un único cuadrante de la OFM. Para el equipo utilizado para este protocolo, un máximo de 4 ratones individuales podría ser rastreado utilizando cada cuadrante del laberinto. Si la utilización de un laberinto de múltiples recinto, después de realizar el primer tema del ratón en su cuadrante definido, coloque los ratones restantes en su respectivo cuadrante laberinto para el seguimiento de análisis. A los fines de este protocolo, más instrucción será específico para un solo cuadrante del laberinto. <ol><li> Llevar los ratones en sus jaulas de su habitación vivienda en la sala de pruebas. Permita que los ratones se aclimate a la sala de procedimientos por un mínimo de 30 minutos antes de comenzar la prueba. </li><li> Quitar una sola mOuse desde la jaula de alojamiento agarrando suavemente su cola y colocar el ratón en el centro del laberinto campo abierto, mientras que al mismo tiempo activar el software SMART con un simple clic en el botón Inicio para comenzar a rastrear el movimiento del ratón. Es normal que el ratón para desplazarse inmediatamente a las paredes de la periferia del laberinto y el momento de la liberación y el seguimiento de la captura del ratón debe coincidir para registrar este movimiento. </li><li> Permitir el movimiento libre e ininterrumpido del ratón asignatura a lo largo del respectivo cuadrante del laberinto para un único período de 10 minutos durante los cuales, el software de seguimiento registrará movimiento (Figura 3). </li><li> Al final del período de prueba, recoge el tema del ratón suavemente, de sacarlo del laberinto y devolverlo a su jaula. </li><li> Antes de limpiar el laberinto, contar visualmente los pellets boli fecales presentes en el laberinto y anote los números para su posterior análisis manualmente. </li><li> Quite todos los sedimentos fecales y limpie todos los puntos de la micción. Rocíe el piso y las paredes del cuadrante laberinto con etanol al 95% y limpie con una toalla de papel limpia. Deje que la solución de etanol que se seque completamente antes de probar otros ratones. </li><li> Repetir el procedimiento con el siguiente ratón. </li></ol> 4. Medición y Análisis de Comportamiento Durante el Procedimiento de Pruebas NOTA: Para la medición, tres aspectos de la conducta campo abierto se caracterizan fácilmente utilizando este protocolo (véase el debate). Una breve instrucción sobre cómo acceder a estas medidas en el software de seguimiento de vídeo sigue. <ol><li> Desde la pantalla principal del software SMART, mover el cursor a la ficha y solo clic &quot;zonas&quot; &quot;Definición&quot; para abrir el Editor de Zona. </li><li> Siga las instrucciones detalladas en el Manual del usuario del software SMART no definir zonas o rejillas de la superposición en los caminos de seguimiento. Aquí, el software se utiliza para definir una cuadrícula de 10 cm cuadrados que cubren el suelo del laberinto 5 x 5 ( <strong&gt; Figura 4). Asegúrese de guardar el archivo de zona creada antes de cerrar el Editor de Zona. </li><li> Desde la pantalla principal del software SMART, mover el cursor a la pestaña &quot;Análisis&quot; y solo clic para abrir la ventana de Análisis de Datos. </li><li> Mueva el cursor a la ficha &quot;Archivo&quot; y abra el archivo Zona creado anteriormente. </li><li> Mueva el cursor a la pestaña &quot;Configuración&quot; y abra la opción &quot;Análisis de pista&quot;. Esto abrirá la ventana &quot;Configuración de análisis de un solo sujeto&quot;. <ol><li> Mueva el cursor a la pestaña &quot;Standard&quot; y mueva el parámetro &quot;distancia recorrida&quot; en el cuadro Parámetros Disponible (lado izquierdo) a la caja de parámetros incluidos (lado derecho). </li><li> Mueva el cursor a la pestaña &quot;Zonas Transiciones&quot; y mover todos los parámetros adecuados para el cuadro Parámetros Incluido como anteriormente. </li><li> Asegúrese de que la casilla de &quot;Full Track&quot; está marcada en la parte inferior de la ventana. </li><li> Move el cursor al botón OK y cierre la ventana de configuración de análisis de una sola materia. NOTA: En función del análisis que desea realizar, muchas otras opciones se pueden elegir en esta ventana para extraer datos de los análisis. Lea el manual del usuario detallado de su programa específico para determinar qué parámetros son los más importantes para el análisis de sus datos. </li></ol></li><li> En la ficha Archivo en la parte superior izquierda de la ventana del programa, abra la ventana sola pista tema y colocar una marca de verificación junto a todas las pistas están analizando. Mueva el cursor hasta el botón Marca de Verificación en la parte superior de la ventana y haga clic en para cerrar la ventana Track Explorer. </li><li> Mueva el cursor sobre el botón &quot;Go&quot; en la ventana de Análisis de Datos y solo clic para iniciar el análisis de los datos de la pista. </li><li> Análisis de datos se pueden enviar ya sea como archivos de texto ASCII o puede ser exportado directamente en una hoja de cálculo Excel. Utilice las herramientas de salida del programa de software que está utilizando para dar salida al data para su propio uso. NOTA: la distancia total recorrida y el tiempo empleado en las zonas indicadas se emitirá siguiendo los pasos de análisis de datos antes mencionados. Una vez más, se hace hincapié en que los pasos para llegar a estas mediciones representadas aquí será diferente dependiendo del software de usuario utilizado. Pero los propios datos y la interpretación de los resultados deben ser independientes similar del programa de software utilizado. También vale la pena señalar que el sesgo de administrador de la prueba se elimina de este protocolo ya que todos los datos recogidos se cuantifica datos medidos por el software y no el administrador. Por lo tanto no hay ningún elemento calificable de los datos recogidos tal como se describe. </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.

El Open Field Maze es una de las plataformas más utilizadas en los estudios de comportamiento animal. Una serie de importantes parámetros convencionales y etológicas 2,4 puede ser recogida y analizada durante la realización de la OFM. Estos datos permiten al investigador para medir los comportamientos que van desde la actividad general de la locomotora a comportamientos emocionales relacionados con la ansiedad 8. Sin embargo, el uso de OFM no está exenta de defectos. Un problema confuso es la ...

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><tbody><tr><td>Multi Unit Open Field Test</td><td>San Diego Instruments, Inc.</td><td>White 7001-0354</td><td>Any single or multi unit open field maze can be used</td></tr><tr><td>SMART DT Tracking Software</td><td>PanLab/Harvard Apparatus</td><td>76-0695</td><td>Any tracking software can be utilized with this protocol</td></tr><tr><td>Sony 990x Video Camera Recorder</td><td>Sony</td><td>CCD-TRV328</td><td>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.

El número promedio de individuos por cepa de ratones ensayados en la mayoría de los casos es de aproximadamente 20 para generar suficiente relevancia estadística. Sin embargo, este número puede estar en el intervalo de 8-30 dependiendo de la disponibilidad del ratón. Dependiendo de la medición o comparaciones requeridas, también es favorable para utilizar los sujetos de la misma edad. La primera y posiblemente más importante parámetro específico de medir en el campo Maze Open es la...

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.

El uso del campo Maze abierto para medir el comportamiento locomotor y la ansiedad-como en ratones

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

Universidad San Sebastian

Research

JoVE Journal

Behavior

72.4K vistas.  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.

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

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.

Los callejones de prueba sucesiva de ansiedad en ratones y ratas

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.  ...

Investigación

Comportamiento

Low-stress Route Learning Using the Lashley III Maze in Mice

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other aversive stimuli to motivate task acquisition. As animals age, sensory modalities deteriorate. For example, many strains of mice develop hearing deficits or cataracts. Changes in the sensory systems required to guide mice during task acquisition present potential confounds in interpreting learning changes in aging animals. Moreover, the use of aversive stimuli to motivate animals to learn tasks is potentially confounding when comparing mice with differential sensitivities to stress. To minimize these types of confounding effects, we have implemented a modified version of the Lashley III maze. This maze relies on route learning, whereby mice learn to navigate a maze via repeated exposure under low stress conditions, e.g. dark phase, no food/water deprivation, until they navigate a path from the start location to a pseudo-home cage with 0 or 1 error(s) on two consecutive trials. We classify this as a low-stress behavior test because it does not rely on aversive stimuli to encourage exploration of the maze and learning of the task. The apparatus consists of a modular start box, a 4-arm maze body, and a goal box. At the end of the goal box is a pseudo-home cage that contains bedding similar to that found in the animal&#x2019;s home cage and is specific to each animal for the duration of maze testing. It has been demonstrated previously that this pseudo-home cage provides sufficient reward to motivate mice to learn to navigate the maze1. Here, we present the visualization of the Lashley III maze procedure in the context of evaluating age-related differences in learning and memory in mice along with a comparison of learning behavior in two different background strains of mice. We hope that other investigators interested in evaluating the effects of aging or stress vulnerability in mice will consider this maze an attractive alternative to behavioral tests that involve more stressful learning tasks and/or visual cues.

The Lashley III maze is a route-learning task that does not rely on aversive stimuli or visual cues. It is thus a highly attractive option for evaluating learning and memory, especially in aging mice or otherwise where stress is a consideration.

De baja tensión Ruta de Aprendizaje Usando el laberinto Lashley III en ratones

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other ...

Neurociencias

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 Maze estrategias de ensayo con pequeñas y grandes modelos de roedores

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 ...

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 y Aprendizaje de respuesta en el Abierto de 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 ...

Novel Object Exploration as a Potential Assay for Higher Order Repetitive Behaviors in Mice

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals. RRBs are commonly split into lower-order and higher-order components, with lower order RRBs consisting of motor stereotypies and higher order RRBs consisting of perseverative and sequencing behaviors. Higher order RRBs are challenging to model in mice. Current assays for RRBs in mice focus primarily on the lower order components, making basic biomedical research into potential treatments or interventions for higher-order RRBs difficult. Here we describe a new assay, novel object exploration. This assay uses a basic open-field arena with four novel objects placed around the perimeter. The test mouse is allowed to freely explore the arena and the order in which the mouse investigates the novel objects is recorded. From these data, patterned sequences of exploration can be identified, as can the most preferred object for each mouse. The representative data shared here and past results using the novel object exploration assay illustrate that inbred mouse strains do demonstrate different behavior in this assay and that strains with elevated lower order RRBs also show elevated patterned behavior. As such, the novel object exploration assay appears to possess good face validity for higher order RRBs in humans and may be a valuable assay for future studies investigating novel therapeutics for ASD.

Higher order restricted, repetitive behaviors (RRBs) disrupt the lives of affected individuals. These behaviors are challenging to model in rodents, making basic biomedical research into potential treatments or interventions for RRBs difficult. Here we describe novel object exploration as a potential assay for higher order RRBs in mice.

Exploración de objetos novela como un ensayo de potencial de mayor orden repetitivo Comportamientos en ratones

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals.  RRBs are ...

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.

Laberinto elevado Plus para ratones

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 ...

Biología

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.

Analizar el aprendizaje espacial y el comportamiento Prosocial en los ratones con el laberinto de Barnes y paradigmas de la damisela en peligro

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

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and dysfunctions in mood and cognition are common and often shared symptoms in neurodegenerative diseases. Shown here is a behavioral test battery for motor, mood, and cognition, which can be repeatedly tested in a longitudinal study. This battery assesses the overall behavioral phenotype in mice by examining each domain of behavior with at least two independent well-accepted tests (i.e., open-field test and rotarod test for motor function, social interaction test, elevated plus maze test, and forced swim test for emotional function, and Morris water maze test and novel object recognition test for cognitive function). Therefore, this sensitive and comprehensive test battery is a powerful tool for the study of behavioral alternation in neurodegeneration.

A comprehensive behavioral test battery of motor skills, mood&#8212;including social interaction, depression, and anxiety&#8212;and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.

Una batería de pruebas de comportamiento para la evaluación repetida de la motricidad, humor y cognición en ratones

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and ...

Investigating Migraine-Like Behavior Using Light Aversion in Mice

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as photophobia. Whilst it is impossible to confirm that a mouse is experiencing migraine, light aversion can be used as a behavioral surrogate for the migraine symptom of photophobia. To test for light aversion, we utilize the light/dark assay to measure the time mice freely choose to spend in either a light or dark environment. The assay has been refined by introducing two critical modifications: pre-exposures to the chamber prior to running the test procedure and adjustable chamber lighting, permitting the use of a range of light intensities from 55 lux to 27,000 lux. Because the choice to spend more time in the dark is also indicative of anxiety, we also utilize a light-independent anxiety test, the open field assay, to distinguish anxiety from light-aversive behavior. Here, we describe a modified test paradigm for the light/dark and open field assays. The application of these assays is described for intraperitoneal injection of calcitonin gene-related peptide (CGRP) in two mouse strains and for optogenetic brain stimulation studies.

Rodents are not able to report migraine symptoms. Here, we describe a manageable test paradigm (light/dark and open field assays) to measure light aversion, one of the most common and bothersome symptoms in patients with migraines.

Investigando el comportamiento similar a la migraña utilizando la aversión a la luz en ratones

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as ...

Validación de un modelo murino del síndrome de Angelman mediante pruebas conductuales

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.

Author Spotlight: Una batería de pruebas conductuales altamente reproducibles para validar un modelo murino de síndrome de Angelman 

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

Caracterización conductual de un modelo de ratón con 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 ...