This work is supported by a Baylor University Research Council grant and by a research grant from the Epilepsy Foundation.

Los ratones machos adultos utilizados en el experimento siguiente se generaron y se alojaron en la Universidad de Baylor en una temperatura ambiente de 22 ° C, con una luz de 14 horas y 10 horas oscuridad (20:00-6:00 hr) ciclo diurno. Los ratones tuvieron acceso ad libitum a comida y agua. Todos los procedimientos a los ratones estaban en el cumplimiento de los Institutos Nacionales de Salud para el Cuidado y Uso de Animales de Laboratorio y el protocolo de animales fue aprobado por la Universidad de Baylor Cuidado de Animales y el empleo Comisión. Los ratones se pusieron a prueba en el final de la mañana / tarde. Para todos los procedimientos a continuación el investigador debe usar guantes para eliminar olores y evitar la contaminación. 1. Preparación de olores no sociales <ol><li> Preparar los olores no sociales frescas cada día de la prueba. Nota: El investigador debe utilizar guantes para preparar los olores. <ol><li> Rellene 3 tubos de centrífuga con 10 ml de agua cada uno. </li><li> Pipetear 100 l de extracto de plátano en un 15 ml &amp; #160; tubo cónico y 100 l de extracto de almendra en un segundo tubo cónico de 15 ml para una dilución 1: 100. Nota: El tercer tubo 15 ml sólo contendrá agua. </li></ol></li></ol> 2. Preparación de los olores Sociales <ol><li> Preparar olores sociales frescas en las pruebas de la mañana va a producir. Obtener olores sociales a partir de dos jaulas que no han sido limpiados por lo menos durante tres días, y contener un número igual de ratones. Obtener olores sociales de los ratones del mismo sexo que el sujeto de la prueba. Nota: Estos son para ser almacenado a temperatura ambiente y se deben utilizar dentro de 4-6 horas para mantener el olor fuerte y consistente. <ol><li> Determinar cuántos se va a crear el olor sociales hisopos de algodón necesitan: cada ratón está probando se presentará con tres hisopos de olor sociales 1, y tres de olor sociales 2. </li><li> Preparar olores sociales frotando un hisopo de algodón en forma de zigzag a través de la parte inferior de una jaula sucia. Agitar el exceso de ropa de cama de aferrarse a la punta. </li><li> Hisopos de las tiendas en grandes frascos de vidrio con tapa o en vasos cubiertos con parafilm. Designar un contenedor para el olor sociales 1 y un segundo para el olor sociales 2. </li></ol></li><li> Mantener las jaulas de origen olor fuera de la sala de pruebas experimentales. </li></ol> 3. Aclimatación del ratón de prueba <ol><li> Póngase los guantes y pesar el ratón para ser probado y colocar individualmente en una jaula limpia con una tapa de alambre en la parte superior y la botella de agua eliminada. </li><li> Deje que el ratón para aclimatarse a la nueva jaula durante 45 minutos en una habitación aparte de la sala de pruebas. Asegúrese de que no hay olores extraños o ruidos en la habitación. </li><li> Como prueba dura aproximadamente 45 minutos, escalonar los tiempos de aclimatación al poner el ratón junto a ensayar en una jaula limpia para aclimatarse a la derecha antes de probar el ratón actual. </li></ol> 4. Preparación de la sala de pruebas <ol><li> Coloque los hisopos de algodón fresco, cinta de laboratorio, las botellas que contienen hisopos de olores sociales, los tubos de nin sociales olores, una micropipeta, dos temporizadores, y una botella de residuos con una tapa en la sala de pruebas. </li><li> Asegúrese de que la sala de pruebas está razonablemente libre de olores, ruidos fuertes, y de luz brillante. No use colonias o lociones perfumadas. Nota: El observador no debe ser la goma de mascar o caramelos durante el período de pruebas y debe tratar de limitar su exposición a fuertes alimentos de olor. </li></ol> 5. Procedimiento de prueba <ol><li> Traiga el ratón ambientada en su jaula en la sala de pruebas y colocar la jaula en el mostrador a una altura que sea cómoda para su visualización mientras está sentado. </li><li> Presentar cada olor 3 veces seguidas durante 2 minutos cada vez. Utilice el orden de presentación que se pueden encontrar en la Tabla 1. <ol><li> Pipeta 50 l de odorizante en el olor a ensayar a la punta del hisopo de algodón y re-cap la fuente de olor. Levante la parte superior de la jaula de alambre e inserte el extremo sin perfume, de madera de la torunda en la abertura de la botella de agua, empujandog la longitud a través de la parte inferior del cable a la parte superior. Evite tocar la punta de algodón para cualquier cosa, incluyendo los dedos, de modo que se evite la contaminación de olores. </li><li> Continuar para dibujar el hisopo a través de la abertura de la botella de agua hasta aproximadamente 1 pulgada del extremo de algodón cuelga hacia abajo en la jaula. Colocar el extremo de madera de la torunda hacia un lado de la jaula usando cinta de laboratorio. Nota: Tenga cuidado de no dejar caer la tapa de nuevo en la jaula, provocando un fuerte ruido que perturbe el ratón prueba. </li><li> Inmediatamente comenzará la sincronización durante 2 min. Use un temporizador para medir el tiempo de la presentación de 2 minutos. Registre el tiempo acumulado pasado oler el olor durante el juicio 2 min. Cuente comportamiento inhalación del animal utilizando un cronómetro separado. Sniffing activo se define cuando la nariz del animal se orienta hacia la punta de algodón con su nariz ~ 2 cm o más cerca de la punta. </li><li> Tras la finalización de cada ensayo, levantar la tapa de alambre y retire el hisopo de algodón tirando de él hacia abajo a través de la ONUlado inferior de la tapa (esto evita la contaminación de olores mediante la prevención de la punta de algodón de tocar el cable). Sellar el hisopo utilizado en botella de residuos de 500 ml. </li><li> Preparar e insertar la siguiente hisopo y comenzar la sincronización. Deje aproximadamente 1 minuto entre cada intervalo entre ensayos. Continúe hasta que cada aroma se ha presentado tres veces seguidas, durante 2 minutos cada vez. </li></ol></li><li> Una vez que el ratón se ha completado todos los ensayos, vaciar la botella de residuos que contienen los hisopos de algodón usados ​​en un bote de basura fuera de la sala de ensayo. Devuelva el ratón a su jaula. </li></ol> 6. Análisis <ol><li> Analizar los datos para cada grupo por condición olor usando ANOVA de dos vías seguido de Newman-Keuls o una prueba post hoc similar para determinar la habituación significativa (menos tiempo olfateando sucesivas mismos olores), y dishabitutation (más tiempo olfateando un nuevo olor). </li></ol>

<ol>
	<li>Doty, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odor-guided+behavior+in+mammals.">Odor-guided behavior in mammals.</a> Experientia. 42, 257-271 (1986).</li><li>Restrepo, D., Arellano, J., Oliva, A. M., Schaefer, M. L., Lin, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emerging+views+on+the+distinct+but+related+roles+of+the+main+and+accessory+olfactory+systems+in+responsiveness+to+chemosensory+signals+in+mice.">Emerging views on the distinct but related roles of the main and accessory olfactory systems in responsiveness to chemosensory signals in mice.</a> Horm Behav. 46, 247-256 (2004).</li><li>Keverne, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Importance+of+olfactory+and+vomeronasal+systems+for+male+sexual+function.">Importance of olfactory and vomeronasal systems for male sexual function.</a> Physiol Behav. 83, 177-187 (2004).</li><li>Gregg, B., Thiessen, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+of+olfactory+discrimination+of+urines+for+the+Mongolian+gerbil,+Meriones+unguiculatus.">A simple method of olfactory discrimination of urines for the Mongolian gerbil, Meriones unguiculatus.</a> Physiol Behav. 26, 1133-1136 (1981).</li><li>Yang, M., 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=Simple+behavioral+assessment+of+mouse+olfaction.">Simple behavioral assessment of mouse olfaction.</a> Curr Protoc Neurosci. Chapter 8 (Unit 8 24), (2009).</li><li>Pan, Y. W., Kuo, C. T., Storm, D. R., Xia, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inducible+and+targeted+deletion+of+the+ERK5+MAP+kinase+in+adult+neurogenic+regions+impairs+adult+neurogenesis+in+the+olfactory+bulb+and+several+forms+of+olfactory+behavior.">Inducible and targeted deletion of the ERK5 MAP kinase in adult neurogenic regions impairs adult neurogenesis in the olfactory bulb and several forms of olfactory behavior.</a> PLoS ONE. 7, e49622 (2012).</li><li>Wesson, D. W., Levy, E., Nixon, R. A., Wilson, 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=Olfactory+dysfunction+correlates+with+amyloid-beta+burden+in+an+Alzheimer's+disease+mouse+model.">Olfactory dysfunction correlates with amyloid-beta burden in an Alzheimer's disease mouse model.</a> J Neurosci. 30, 505-514 (2010).</li><li>Fadool, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kv1.3+channel+gene-targeted+deletion+produces+'Super-Smeller+Mice'+with+altered+glomeruli,+interacting+scaffolding+proteins,+and+biophysics.">Kv1.3 channel gene-targeted deletion produces 'Super-Smeller Mice' with altered glomeruli, interacting scaffolding proteins, and biophysics.</a> Neuron. 41, 389-404 (2004).</li><li>Arakawa, H., Arakawa, K., Blanchard, D. C., Blanchard, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scent+marking+behavior+in+male+C57BL/6J+mice:+sexual+and+developmental+determination.">Scent marking behavior in male C57BL/6J mice: sexual and developmental determination.</a> Behav Brain Res. 182, 73-79 (2007).</li><li>Baum, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+differentiation+of+pheromone+processing:+links+to+male-typical+mating+behavior+and+partner+preference.">Sexual differentiation of pheromone processing: links to male-typical mating behavior and partner preference.</a> Horm Behav. 55, 579-588 (2009).</li><li>Ferguson, J. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+amnesia+in+mice+lacking+the+oxytocin+gene.">Social amnesia in mice lacking the oxytocin gene.</a> Nat Genet. 25, 284-288 (2000).</li><li>Yang, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+sociability+in+BTBR+T+tf/J+mice+is+independent+of+partner+strain.">Low sociability in BTBR T+tf/J mice is independent of partner strain.</a> Physiol Behav. 107, 649-662 (2012).</li><li>Best, J. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-associative+learning+in+larval+zebrafish.">Non-associative learning in larval zebrafish.</a> Neuropsychopharmacology. 33, 1206-1215 (2008).</li><li>Wolman, M. A., Jain, R. A., Liss, L., Granato, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+modulation+of+memory+formation+in+larval+zebrafish.">Chemical modulation of memory formation in larval zebrafish.</a> Proc Natl Acad Sci U S A. , (2011).</li><li>Sorge, R. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+exposure+to+males,+including+men,+causes+stress+and+related+analgesia+in+rodents.">Olfactory exposure to males, including men, causes stress and related analgesia in rodents.</a> Nat Methods. 11, 629-632 (2014).</li></ol>

The authors have nothing to disclose.

Los resultados proporcionados en este documento son óptimas para los ratones. Los ratones muestran una fuerte respuesta a un nuevo olor, a continuación, se habitúan rápidamente. Uno de los pasos clave en este método es la preparación de los olores. El investigador tiene que tener mucho cuidado para aislar los olores entre sí para evitar la contaminación. Otro aspecto clave es la presentación de la esponja de algodón. El investigador tiene que solucionar la mejor ubicación de la punta de algodón por lo que el...

Los ratones son dependientes de olfato para navegar nuevos entornos, la búsqueda de alimentos, para el reconocimiento de otras personas, y el comportamiento sexual 1-3. Es esencial que los investigadores determinar si o no los animales experimentales tienen un sentido del olfato funcionamiento antes de la administración de las pruebas de comportamiento que implican la comida, la interacción social, o cualquier olor destinadas a provocar una respuesta desde el ratón. La anosmia o la incapacidad para distinguir entre diferentes olores, podría dar lugar a falsos positivos o negativos en una amplia variedad de paradigmas de comportamiento, por lo que la capacidad de un ratón para detectar y distinguir los olores se deben establecer antes de realizar otros tipos de pruebas de comportamiento. La prueba de habituación / deshabituación olfativa fue descrita por primera vez en la década de 1980 4. La prueba ha sido adaptado para su uso en los ratones por los Dres. Mu Yang y Jacqueline Crawley 5. Esta es una prueba sencilla y de bajo costo que permite al investigador thisblecer que un ratón puede detectar y diferenciar entre los olores. Además de probar el olfato, esta prueba permite al investigador para observar el comportamiento general del ratón y debe hacerse temprano en un régimen de pruebas. Observaciones cualitativas respecto a la locomoción del ratón, los signos de ansiedad, nivel de actividad, y la respuesta a los olores sociales frente a los olores de los alimentos puede ser señal de nuevas áreas en las que la prueba podría llevarse a cabo. En esta prueba, hisopos de algodón humedecido en varios olores se presentan a un ratón de tres veces seguidas. Con cada presentación repetida de un olor, el ratón se habitúan a la esponja de algodón, gastando menos tiempo a investigar con cada presentación posterior. Cuando se presenta un nuevo olor, deshabituación ocurre, y un ratón típico pasar más tiempo investigando el hisopo, lo que indica que puede discriminar entre los olores actuales y los anteriores 5. Esta prueba se administra a un ratón en un momento e incluye un acc 45 minperiodo limation seguido por 45 min de la prueba. Aunque esta prueba es fácil de llevar a cabo que se puede utilizar para investigar cuestiones sofisticadas sobre el sistema olfativo del ratón. Otras pruebas populares del olfato, como la prueba de alimentos enterrados, simplemente establecer la presencia o ausencia de la anosmia. Sin embargo, la prueba de discriminación y la habituación olfativa permite al investigador determinar que un ratón no sólo tiene la capacidad de detectar olores pero puede discriminar entre diferentes olores. El patrón de habituación y deshabituación se ha utilizado para establecer que los nuevos mutantes pueden discriminar entre los olores 6,7. En un estudio sorprendente, Fadool y sus colegas utilizaron mezclas complejas de olores para mostrar que los ratones con deleción del gen de orientación del canal Kv1.3 son &quot;súper smellers&quot; que pueden discriminar olores muy similares mejor que los ratones normales 8. Al examinar un nuevo modelo de ratón knockout es útil establish la presencia de un comportamiento normal para tareas sensoriales básicos. Cuando se hace temprano en un régimen de pruebas, la discriminación de olores y prueba de habituación da un investigador la oportunidad de observar cualquier comportamiento inusual. Estas observaciones podrían evitar resultados positivos o falsos negativos en las pruebas posteriores que son atribuibles a la confusión características del mutante. Como investigadores continúan investigando el comportamiento social la necesidad de verificar la función olfativa básico será cada vez más importante. Además de examinar el olfato en líneas mutantes, esta prueba se puede utilizar para abordar cuestiones específicas, como si un tratamiento farmacológico aumenta específicamente la respuesta de un animal a los estímulos de olor sociales o si aumenta su respuesta a todos los estímulos de olor.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Mouse cage</td>
			<td>Allentown</td>
			<td>Standard mouse cage</td>
			<td></td>
		</tr>
		<tr>
			<td rowspan="2">Wire lid</td>
			<td rowspan="2">Allentown</td>
			<td>BCU Mouse WBL 2500</td>
			<td rowspan="2"></td>
		</tr>
		<tr>
			<td></td>
		</tr>
		<tr>
			<td>Bedding</td>
			<td>Harlan</td>
			<td>7090 Sani-Chips</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swabs</td>
			<td>VWR</td>
			<td>89031-270</td>
			<td>6&#8221; wooden handle</td>
		</tr>
		<tr>
			<td>Banana extract</td>
			<td>McCormick</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Almond extract</td>
			<td>McCormick</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Laboratory tape</td>
			<td>VWR</td>
			<td>89098-062</td>
			<td></td>
		</tr>
		<tr>
			<td>Stop watch</td>
			<td>VWR</td>
			<td>62374-000</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrile gloves</td>
			<td>VWR</td>
			<td>82026</td>
			<td></td>
		</tr>
		<tr>
			<td>Timing device</td>
			<td>VWR</td>
			<td>61161-350</td>
			<td></td>
		</tr>
		<tr>
			<td>15 mL conical tubes</td>
			<td>VWR</td>
			<td>89003-294</td>
			<td></td>
		</tr>
		<tr>
			<td>2 L beakers&#160;</td>
			<td>Pyrex</td>
			<td align="right">1003</td>
			<td></td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>Parafilm</td>
			<td>PM-992</td>
			<td>4&#8221; x 250&#8217;</td>
		</tr>
		<tr>
			<td>1 L bottle with cap</td>
			<td>VWR</td>
			<td>89000-240</td>
			<td></td>
		</tr>
	</tbody>
</table>

testing for odor discrimination and habituation in mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the mouse can discriminate between non-social odors and social odors, whether the mouse habituates to a repeatedly presented odor, and whether the mouse demonstrates dishabituation when presented with a novel odor. Since many social behavior tests measure the experimental animal&#8217;s response to a familiar or novel mouse, false positives can be avoided by establishing that the animals can detect and discriminate between social odors. There are similar considerations in learning tests such as fear conditioning that use odor to create a novel environment or olfactory cues as an associative stimulus. Deficits in the olfactory system would impair the ability to distinguish between contexts and to form an association with an olfactory cue during fear conditioning. 
In the odor habitation/dishabituation test, the mouse is repeatedly presented with several odors. Each odor is presented three times for two minutes. The investigator records the sniffing time directed towards the odor as the measurement of olfactory responsiveness. A typical mouse shows a decrease in response to the odor over repeated presentations (habituation). The experimenter then presents a novel odor that elicits increased sniffing towards the new odor (dishabituation). After repeated presentation of the novel odor the animal again shows habituation. This protocol involves the presentation of water, two or more non-social odors, and two social odors. In addition to reducing experimental confounds, this test can provide information on the function of the olfactory systems of new knockout, knock-in, and conditional knockout mouse lines.

Siguiendo el protocolo descrito, los resultados se registraron utilizando diez adultos (día postnatal 90-120) macho C57BL / 6J y ocho macho cepa FVB mixta de fondo (día postnatal 50-70). La prueba se llevó a cabo por un solo investigador a través de múltiples días. Cada punto en los gráficos representa el tiempo medio dedicado a investigar un olor. Cuando el hisopo de algodón humedecido en agua se introdujo por primera vez, los ratones pasan una gran cantidad de tiempo investigando e...

Watch this Scientific Journal Video about Testing for Odor Discrimination and Habituation in Mice at JoVE.com

Testing for Odor Discrimination and Habituation in Mice

This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor.

Las pruebas para detectar olor Discriminación y habituación en ratones

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the ...

testing-for-odor-discrimination-and-habituation-in-mice

Research

JoVE Journal

Behavior

17.5K vistas.  Baylor University. This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor. 

Video: Testing for Odor Discrimination and Habituation in Mice

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association between environmental cues and aversive experiences. In this test, mice are placed into a conditioning chamber and are given parings of a conditioned stimulus (an auditory cue) and an aversive unconditioned stimulus (an electric footshock). After a delay time, the mice are exposed to the same conditioning chamber and a differently shaped chamber with presentation of the auditory cue. Freezing behavior during the test is measured as an index of fear memory. To analyze the behavior automatically, we have developed a video analyzing system using the ImageFZ application software program, which is available as a free download at http://www.mouse-phenotype.org/. Here, to show the details of our protocol, we demonstrate our procedure for the contextual and cued fear conditioning test in C57BL/6J mice using the ImageFZ system. In addition, we validated our protocol and the video analyzing system performance by comparing freezing time measured by the ImageFZ system or a photobeam-based computer measurement system with that scored by a human observer. As shown in our representative results, the data obtained by ImageFZ were similar to those analyzed by a human observer, indicating that the behavioral analysis using the ImageFZ system is highly reliable. The present movie article provides detailed information regarding the test procedures and will promote understanding of the experimental situation.

This article presents a protocol for a contextual and cued fear conditioning test using a video analyzing system to assess fear learning and memory in mice.

Contextuales y complementada el condicionamiento del miedo de prueba utilizando un sistema de análisis de vídeo en ratones

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association ...

Investigación

Comportamiento

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 ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

La Junta Agujero Modificado - Medición de Comportamiento, Cognición e interacción social en ratones y ratas

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Testing Sensory and Multisensory Function in Children with Autism Spectrum Disorder

In addition to impairments in social communication and the presence of restricted interests and repetitive behaviors, deficits in sensory processing are now recognized as a core symptom in autism spectrum disorder (ASD). Our ability to perceive and interact with the external world is rooted in sensory processing. For example, listening to a conversation entails processing the auditory cues coming from the speaker (speech content, prosody, syntax) as well as the associated visual information (facial expressions, gestures). Collectively, the &#8220;integration&#8221; of these multisensory (i.e., combined audiovisual) pieces of information results in better comprehension. Such multisensory integration has been shown to be strongly dependent upon the temporal relationship of the paired stimuli. Thus, stimuli that occur in close temporal proximity are highly likely to result in behavioral and perceptual benefits &#8211; gains believed to be reflective of the perceptual system&#39;s judgment of the likelihood that these two stimuli came from the same source. Changes in this temporal integration are expected to strongly alter perceptual processes, and are likely to diminish the ability to accurately perceive and interact with our world. Here, a battery of tasks designed to characterize various aspects of sensory and multisensory temporal processing in children with ASD is described. In addition to its utility in autism, this battery has great potential for characterizing changes in sensory function in other clinical populations, as well as being used to examine changes in these processes across the lifespan.

We describe how to implement a battery of behavioral tasks to examine the processing and integration of sensory stimuli in children with ASD. The goal is to characterize individual differences in temporal processing of simple auditory and visual stimuli and relate these to higher order perceptual skills like speech perception.

Prueba de la función sensorial y multisensorial en niños con Trastorno del Espectro Autista

In addition to impairments in social communication and the presence of restricted interests and repetitive behaviors, deficits in sensory processing are ...

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

Morris Water Maze Prueba: Optimización de ratón Colar y entorno de pruebas

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

A Protocol for Housing Mice in an Enriched Environment

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent studies demonstrate that environmental enrichment decreases adiposity, increases energy expenditure, resists diet induced obesity, and causes cancer remission and inhibition in mice. EE typically consists of larger living space, a variety of &#8216;toys&#8217; to interact with, running wheels, and can include a number of other novel environmental changes. All of this fosters a more complex social engagement, cognitive and physical stimulations. Importantly, the toy location and type of toy is changed regularly, which encourages the mice to adapt to a frequently changing and novel environment. Many variables can be manipulated in EE to promote health effects in mice. Thus these approaches are difficult to control and must be properly managed to successfully replicate the associated phenotypes. Therefore, the goal of this video is to demonstrate how EE is properly set up and maintained to assure a complex, challenging, and controlled environment so that other researchers can easily reproduce the protective effects of EE against obesity and cancer.

Environmental enrichment for mice requires a complex and challenging setup, as well as comprehensive husbandry and handling techniques to assure robust metabolic and anti-cancer effects in the mice. This protocol provides detailed procedures to reproduce the above mentioned effects in mice.

Un Protocolo para ratones Viviendas en un ambiente enriquecido

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent ...

Simple and Computer-assisted Olfactory Testing for Mice

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected with aging and is a strong predictor of neurodegenerative diseases. Thus, olfaction testing is used as a non-invasive diagnostic method to detect neurological deficits early on. In order to understand the mechanisms underlying olfactory network susceptibility, olfactory research in rodents has gained momentum in the past decade.
Here, we present a very simple, time efficient and reproducible olfactory testing method of innate odor perception and sensitivity in mice without the need of any prior food or water restriction. The tests are performed in a familiar environment to the mice, require only the scents and a 2 min session of odorant exposure. The analysis is performed, post-hoc, using computer-assisted commands on ImageJ and can be, therefore, carried out from start to end by one researcher.
This protocol does not require any special hardware or setup and is indicated for any laboratory interested in testing olfactory perception and sensitivity.

We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing.

Simple y asistida por ordenador de pruebas olfativas para Ratones

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected ...

Testing for Metacognitive Responding Using an Odor-based Delayed Match-to-Sample Test in Rats

Metamemory involves the cognitive ability to assess the strength of one&#39;s memories. To explore the possibility of metamemory in non-human animals, numerous behavioral tasks have been created, many of which utilize an option to decline memory tests. To assess metamemory in rats, we utilized this decline-test option paradigm by adapting previous visual delayed-match-to-sample tests (DMTS)1,2 developed for primate species to an odor-based test suitable for rodents. First,&#160;rats are given a sample to remember by digging in a cup of scented sand. After a delay, the rat is presented with four distinctly scented cups, one of which contains the identical scent experienced during the sample; if this matching cup is selected, then the rat obtains a preferred, larger reward. Selection of any of the other three non-matching sand-filled scented cups results in no reward. Retention intervals are individually titrated such that subjects perform between 40 and 70% correct, therefore ensuring rats sometimes remember and sometimes forget the sample. Here, the operational definition of metamemory is the ability to distinguish between the presence and absence of memory through behavioral responding. Towards this end, on two-thirds of trials, a decline option is presented in addition to the four choice cups (choice trials). If the decline-test option- an unscented colored sand cup, is selected, the subject receives a smaller less-preferred reward and avoids the memory test. On the remaining third of trials, the decline-test option is not available (forced trials), causing subjects to guess the correct cup when the sample is forgotten. On choice tests, subjects that know when they remember should select the decline option when memory is weak rather than take the test and choose incorrectly. Therefore, significantly higher performance on chosen tests as compared to forced memory tests is indicative of the adaptive use of the decline-test response and metacognitive responding.

This protocol describes a method for investigating the possibility of metamemory, or memory awareness, in rodents. The odor-based delayed-matching-to-sample paradigm is a novel, ecologically-relevant behavioral test useful for determining the extent to which rodents can adaptively respond based on cognitively monitoring the strength of their memory states.

Pruebas para responder metacognitivas usando un olor basado en diferido partido-a-Sample Test en ratas

Metamemory involves the cognitive ability to assess the strength of one&#39;s memories. To explore the possibility of metamemory in non-human animals, ...

Measuring Active and Passive Tameness Separately in Mice

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two behavioral components: a reluctance to avoid humans (passive tameness) and a motivation to approach humans (active tameness). To quantify these components in mice, we previously developed behavioral tests for active tameness, passive tameness, and the willingness to stay on a human hand, each designed to be completed within 3 min. The data obtained were used for selective breeding, with a large number of mice analyzed per generation. The active tameness test measures the movement of the mouse toward a human hand and the contact it engages in. The passive tameness test measures the duration of time that a mouse tolerates human touch. In the stay-on-hand test, a mouse is placed on a human hand and touched slowly using the thumb of that hand; the duration of time that the animal remains on the hand is measured. Here, we describe the test set-up and apparatus, explain the procedures, and discuss the appropriate data analysis. Finally, we explain how to interpret the results.

Tameness in animals includes a reduction of avoidance responses to humans (passive tameness) and an increase in active approaches to humans (active tameness). Here, we describe detailed protocols for three behavioral tests (active tameness, passive tameness, and stay-on-hand tests) to separately measure the active and passive tameness of mice.

Medición activa y pasiva mansedumbre por separado en ratones

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two ...

A Wind Tunnel for Odor Mediated Insect Behavioural Assays

Olfaction is the most important sensory mechanism by which many insects interact with their environment and a wind tunnel is an excellent tool to study insect chemical ecology. Insects can locate point sources in a three-dimensional environment through the sensory interaction and sophisticated behavior. The quantification of this behavior is a key element in the development of new tools for pest control and decision support. A wind tunnel with a suitable flight section with laminar air flow, visual cues for in-flight feedback and a variety of options for the application of odors can be used to measure complex behaviour which subsequently may allow the identification of attractive or repellent odors, insect flight characteristics, visual-odor interactions and interactions between attractants and odors lingering as background odors in the environment. A wind tunnel holds the advantage of studying the odor mediated behavioural repertoire of an insect in a laboratory setting. Behavioural measures in a controlled setting provide the link between the insect physiology and field application. A wind tunnel must be a flexible tool and should easily support the changes to setup and hardware to fit different research questions. The major disadvantage to the wind tunnel setup described here, is the clean odor background which necessitates special attention when developing a synthetic volatile blend for field application.

Here, we describe the construction and use of a wind tunnel for odor mediated behavioural assays with insects. The wind tunnel design facilitates the release of odor sources by several methods, with and without visual stimuli. Wind tunnel experiments are important methods to identify behaviorally active volatile chemicals.

Un túnel de viento para olor mediada por insectos ensayos conductuales

Olfaction is the most important sensory mechanism by which many insects interact with their environment and a wind tunnel is an excellent tool to study ...