Este trabajo fue apoyado por una subvención-en-Ayudas a JSP becarios a AA; por subvenciones-en-Ayudas a la Investigación Científica en Áreas innovadoras para JSP Fellows (Nº 4501 y Nº 25132712) a los conocimientos tradicionales; y con una donación proyecto de investigación otorgado por la Universidad de Azabu. Figura 2 se reimprime de Kihara, T., Harada, T., y Koshida, fabricación y características de silicio nanocristalino inducidos térmicamente emisores de ultrasonido compatible con la oblea N.. En: Los sensores y actuadores A: físicas, volumen 125, Elsevier, p. 426, (2006), con permiso de Elsevier.

<html> Todos los procedimientos fueron aprobados por el Comité de Ética de la Universidad de Azabu. Todos los experimentos se llevaron a cabo en una cámara insonorizada. 1. Preparación Animal <ol><li> Machos para grabación <ol><li> Obtener ratones machos sexualmente maduros con experiencia en el apareamiento. </li></ol></li><li> Los sujetos femeninos <ol><li> Obtener ratones hembra virgen que han sido alojados con 2 a 5 compañeros de camada por jaula (generalmente 8 - 12 semanas de edad). </li><li> Obtener frotis vaginal a diario para determinar la fase del ciclo estral antes de la prueba, según McLean 21. Se debe tener cuidado para minimizar la estimulación vaginal para evitar pseudopreñez. </li><li> Tinción de los frotis con Giemsa, y determinar la fase del ciclo estral en base a la presencia o ausencia de leucocitos, células epiteliales cornificadas y células epiteliales nucleadas, de acuerdo con Nelson 22. </li></ol></li></ol> 2. Dispositivos (Figura 1) _content &quot;&gt; <img alt="Figura 1" src="/files/ftp_upload/53074/53074fig1.jpg" /> Figura 1. Esquema de la Dos-elección Box Prueba y Dispositivos. Los ratones pueden acceder a salas A y B a través de las pequeñas puertas situadas entre ellos y la zona neutral. La caja de dos elección y dos emisores de ultrasonido se coloca en el suelo de una cámara insonorizada. El micrófono está suspendido dentro de la cámara a prueba de sonido. Abreviatura:. PC, computadora personal <a href="https://www.jove.com/files/ftp_upload/53074/53074fig1large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <ol><li> Para Grabación <ol><li> Conecte un micrófono de condensador de ultrasonidos a un PC mediante un convertidor de USB A / D. </li><li> Ajuste el nivel de ganancia del convertidor A / D a &quot;7&quot;. </li><li> Datos acústicos de Registros en archivos de formato de 16 bits con una frecuencia de muestreo de 400 kHz utilizando el software de captura de audio. </li></ol></li><li> Para la reproducción <ol><li> Conecte un convertidor D / A a la PC usandouna interfaz USB. </li><li> Conecte dos amplificadores al convertidor D / A través de un atenuador. (Este último dispositivo sirve para evitar la sobrecarga.) </li><li> Conectar un emisor de ultrasonido para cada amplificador. </li></ol></li></ol> 3. USV grabación <ol><li> Coloque un ratón macho en una pequeña (17 x 10 x 11 cm) o (21 x 14 x 13 cm) jaula de tamaño mediano con un agujero de 6 cm de diámetro en la pared lateral y el agujero cubierto con una malla de alambre de 0,5 cm. Cuando se utiliza una jaula de tamaño mediano, la construcción de una partición a través del centro de la jaula para reducir el área. </li><li> No use ropa de cama en la jaula para evitar ruidos susurrantes que podrían contaminar las grabaciones. </li><li> Coloque un micrófono al lado de la malla. </li><li> Comience monitoreo vocalización ultrasónica con la configuración de la grabación se muestra en el paso 2.1. </li><li> Colocar un ratón hembra sexualmente maduros, ya sea en metaestro o diestro en la jaula con el ratón macho. </li><li> USVs récord para 3-5 min. </li></ol> 4. Test Box <ol><li> Construir la caja de prueba utilizando tablero de acrílico (Figura 1). Nota: La caja de prueba (35 x 20 y 20 x cm de alto) se divide en tres compartimentos: una zona neutra (15 x 20 cm), y salas A y B (20 x 10 cm, cada uno). Habitaciones A y B tienen cada uno un agujero de 4 cm de diámetro que está cubierto con una malla de alambre de 0,5 cm. Los agujeros de malla y están situados en cada extremo de habitaciones A y B. Hacer puertas (5 x 5 cm cada uno) entre la zona neutral y habitaciones A y B. </li><li> Para evitar la pérdida de sonido aislar cada emisor de ultrasonido, excepto por el lado orientado hacia la malla, usando placas de caucho, sellándolos a la caja alrededor del perímetro de cada malla. </li><li> Lugar sellos de plomo en ambos lados de la pared entre las habitaciones A y B para el aislamiento acústico. </li></ol> 5. Preparación para la reproducción de sonido <ol><li> Creación de sonidos de reproducción <ol><li> Captura de un segmento de 20 segundos de USV del archivo grabado utilizando el software de edición de audio digital. </li><li> Seleccione y captar el fondo sinise (0,35 segundos) desde el archivo segmento sec USV 20. Cree un archivo de ruido de fondo 20 seg repitiendo el segmento de ruido. </li><li> Para reproducir los dos archivos de sonido simultáneamente, exportarlos como un solo archivo de sonido estéreo, con un archivo de la &quot;oreja izquierda&quot; y el otro archivo para el &quot;oído derecho.&quot; </li><li> Filtra los archivos de sonido utilizando como filtro de paso alto con un corte de 40 kHz. </li><li> Reduzca el ruido con la función de reducción de ruido en el software de post-procesamiento. Seleccione y capturar un segmento del archivo que contiene USVs como el perfil de reducción de ruido, y ejecutar la reducción de ruido con el nivel 40. </li></ol></li><li> Calibración de los sonidos de reproducción <ol><li> Asegúrese de que el espacio entre los emisores de ultrasonido y el micrófono es de 10 cm. </li><li> Para calibrar los niveles de presión de sonido de los emisores de ultrasonido, realizar monitoreo ultrasónica usando la configuración descrita en el paso 2.1. Medir el nivel de presión de sonido por el micrófono en decibelios (dB). </li><li> Using el atenuador y los amplificadores, ajustar el volumen de los USVs generadas por los emisores de ultrasonido para el mismo nivel de presión de sonido como los USVs masculinos registrado en el paso 3. </li><li> Al utilizar los dos emisores de ultrasonidos y un archivo de sonido compuesto por USVs y el ruido de fondo, confirmar que los dos emisores de ultrasonidos presentan el mismo nivel de presión de sonido con un sonido de calibración (por ejemplo, un tono puro de 75 kHz) antes de realizar la prueba de comportamiento. </li><li> Al reproducir un archivo de sonido compuesta de dos corrientes de USVs, ajustar los dos niveles de presión de sonido generado al mismo nivel, antes de la prueba. </li></ol></li></ol> 6. Prueba de dos elección <ol><li> Coloque los emisores de ultrasonido detrás de las mallas de la caja de prueba. </li><li> Cerrar las puertas a las salas A y B con un tablero de acrílico, y habituarse al sujeto femenino a la zona neutral durante 30 min. Cubra la caja con tablero de acrílico para evitar que el ratón puede escapar. </li><li> Inicie la grabación de vídeo con un CCCámara D montado en la caja. La cámara (no mostrada en la figura 1) cubre el área de todos los tres compartimientos. </li><li> Retire la puerta y la cubierta a bordo y permitir a la hembra para explorar libremente la caja de prueba, incluyendo las zonas de sonido. </li><li> Una vez que la hembra ha investigado ambas mallas, y ha vuelto a la zona neutral, empezar a jugar un archivo de sonido de 20 segundos en varias ocasiones durante 5 o 10 minutos. </li><li> Llevar a cabo el monitoreo del comportamiento durante 5 o 10 minutos. </li><li> Para reducir señales olfativas deseadas depositado a partir de la materia anterior, limpiar las jaulas de ensayo entre los ensayos con etanol al 70%. </li><li> Cambiar el lugar de emisores de ultrasonidos A y B al azar para evitar el efecto de sesgo lateral inherente hacia salas A o B. </li></ol> 7. Análisis estadísticos <ol><li> El uso de software evento puntuable del comportamiento, análisis de los siguientes parámetros: número total de entradas en cada habitación; duración total de la estancia en cada zona media; duración total de la estancia en cada zona de sonido; yduración total de búsqueda de cada malla en frente del emisor de ultrasonidos. Figura 1 ilustra lo que se entiende por &quot;zona intermedia&quot; y &quot;zona de sonido.&quot; Nota: En los casos en que el ratón se mantiene en la línea divisoria entre las dos zonas (Figura 1, línea discontinua) decidir la ubicación de la dirección de la cabeza. Cuando la cabeza se enfrenta al lado de la malla, se anotó como una estancia en la zona de sonido. </li><li> Para cada parámetro del comportamiento, comparar las respuestas de las habitaciones A y B mediante una prueba de rangos de Wilcoxon firmado-o T-pruebas pareadas con un nivel de significación de 0,05. </li></ol>

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The authors declare that they have no competing interests.

Here, the results of a representative test showed that female mice can discriminate between artificial male USVs and background noise (Figure 3). The conclusion to be drawn from these results is that the discrimination signal is reflected in the duration of stay in the room and sound zone, and in the duration of searching the mesh in the first 5 min of testing, but not in the second 5 min (Figure 3C, E and F). These data indicate that mice become habituated to the playback sounds, possib...

Muchos animales utilizan vocalizaciones para la comunicación intraespecífica. En el ratón Mus musculus, un tipo importante de la señal de comunicación es vocalizaciones ultrasónicas (USVs), que tienen frecuencias superiores a 20 kHz. USVs emitidos por los ratones se consideran un componente de reconocimiento social en hombre-mujer 1.4, hembra-hembra 1, 5 y macho-macho 1, 6 interacciones. USVs también son emitidos por los cachorros cuando están aislados de su madre, lo que aumenta su comportamiento cachorro-recuperación, y por lo tanto PUP supervivencia 7. Aunque muchos informes han analizado y categorizado USVs ratón 8, 9, las respuestas de comportamiento y mecanismos neurales del animal que recibe se han documentado menos 10, 11. Este último es necesario aclarar el significado biológico de las diversas características de USVs. Para revelar estos mecanismos, el experimento de reproducción es un método eficiente. Estudios recientes han revelado que la reproducción femeninaratones son atraídos a USVs 12, y que prefieren USVs de machos que son diferentes de sus padres 13, 14. En este artículo se explica la prueba de reproducción se utiliza para evaluar USV preferencia en ratones. Una caja de prueba de dos elección se desarrolló en el que dos USVs diferentes se pueden reproducir simultáneamente en dos compartimentos de un recinto de ensayo, como se muestra en la Figura 1. Este tipo de caja de prueba evita la contaminación sonora dividiendo el área de prueba en tres sub-cámaras , el uso de paredes de plomo. Los emisores de ultrasonidos se encuentran fuera de cada habitación. En la pared entre las habitaciones y emisores de ultrasonido son agujeros cubiertos con una malla de alambre. Los ratones pueden moverse libremente en las tres habitaciones, y muestran una &quot;búsqueda de la malla&quot; el comportamiento, como para responder a USVs reproducidas por los emisores de ultrasonido. En esta prueba, los ratones se mantienen durante períodos de diferente duración cerca de un emisor de sonido o el otro. Estos parámetros se pueden registrar para obtener un measu sensibles<html> re de la preferencia de sonido. Para reproducir los USVs espalda, emisores termo-acústico de silicio nanocristalino (es decir, &quot;nc-Si emisor&quot;) fueron utilizados como en estudios previos 15-17. Estos dispositivos se componen de un electrodo de película delgada calentador, una capa de silicio nano-poroso, y una oblea de silicio monocristalino. El archivo de sonido digital se convierte en una señal analógica y después se pasa a través del electrodo calefactor. El dispositivo convierte las señales térmicas dependientes de voltaje resultantes en presión sonora significativa con baja distorsión. Este dispositivo es único en que, a diferencia de los generadores de sonido comunes que dependen de las vibraciones mecánicas, se puede reproducir el sonido sin la necesidad de un diafragma. El emisor presenta un nivel de presión sonora plana en las frecuencias de 20 a 160 kHz (Figura 2), y puede reproducir USVs murinos grabados digitalmente con mucha precisión en cuanto a la duración, la frecuencia y el nivel de presión acústica 15, 18, ​​19. ve_content &quot;&gt; En un experimento representativo se muestra en la Figura 3, C57BL / 6 (B6) hembras se les permitió elegir entre BALB / c (BALB) USVs masculinos y ruido de fondo. Además, la Figura 4 muestra la elección de las hembras B6 y BALB entre reproducciones USV simultáneas desde un BALB y un macho B6, según ha informado en un estudio previo 14. Las características de USVs machos difieren entre B6 y BALB cepas 20. Como se muestra por estos resultados, el atractivo de USVs se puede evaluar con el presente Protocolo, en el que los sonidos se registran de un individuo vivo, acústicamente analizados, y se reproducen a otros individuos.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Soundproof chamber</td>
			<td>Muromachi Kikai</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Small cage</td>
			<td>CLEA Japan</td>
			<td>CL-0113-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Middle cage</td>
			<td>CLEA Japan</td>
			<td>CL-0103-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound condenser microphones</td>
			<td>Avisoft Bioacoustics</td>
			<td>CM16/CMPA</td>
			<td></td>
		</tr>
		<tr>
			<td>A/D converter</td>
			<td>Avisoft Bioacoustics</td>
			<td>UltraSoundGate116H</td>
			<td></td>
		</tr>
		<tr>
			<td>Audio software</td>
			<td>Avisoft Bioacoustics</td>
			<td>RECORDER USGH</td>
			<td></td>
		</tr>
		<tr>
			<td>Adobe Audition 3.0 / Audio editing software</td>
			<td>Adobe Systems</td>
			<td>Adobe Audition 3.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Nc-Si emitter</td>
			<td>Original</td>
			<td></td>
			<td>not commercially available but it is planned to be so in near future</td>
		</tr>
		<tr>
			<td>D/A converter</td>
			<td>National Instruments</td>
			<td>NI USB-6251 BNC</td>
			<td></td>
		</tr>
		<tr>
			<td>Attenuator</td>
			<td>Original</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier</td>
			<td>Yamatake</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PC</td>
			<td>Windows 7 professional</td>
			<td></td>
			<td>Intel&#174; core i7-2600K CPU @ 3.4GHz, 8GB RAM, 64-bit operating system</td>
		</tr>
		<tr>
			<td>Event recorder Excel-macro / Event-scoring software</td>
			<td>original</td>
			<td></td>
			<td>Programmed by Naoto Akagawa &#38; Takeru Yamamoto</td>
		</tr>
		<tr>
			<td>CCD Camera</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rubber plates (made of elastomer resin)</td>
			<td>Tokyo bouon</td>
			<td>TI-75BK B4</td>
			<td>Cut them to the proper size http://www.piano-bouon.jp/shopping/?pid=1329272401-447630&#38;ca=6&#38;p=3</td>
		</tr>
		<tr>
			<td>Giemsa&#39;s azur eosin methylene blue solution</td>
			<td>Merck Millipore</td>
			<td>1.09204.0500</td>
			<td></td>
		</tr>
	</tbody>
</table>

determining ultrasonic vocalization preferences in mice using a two-choice playback test

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, sex, condition, and genetic background of the emitting animal. Although many studies have characterized these differences, whether receiver mice can discriminate among objectively different USVs and show preferences for particular sound traits remains to be elucidated. To determine whether mice can discriminate between different characteristics of USVs, a playback experiment was developed recently, in which preference responses of mice to two different USVs could be evaluated in the form of a place preference.
First, USVs from mice were recorded. Then, the recorded USVs were edited, trimmed accordingly, and exported as stereophonic sound files. Next, the USV amplitudes generated by the two ultrasound emitters used in the experiment were adjusted to the same sound pressure level. Nanocrystalline silicon thermo-acoustic emitters were used to play the USVs back. Finally, to investigate the preference of subject mice to selected USVs, pairs of two differing USV signals were played back simultaneously in a two-choice test box. By repeatedly entering a defined zone near an ultrasound emitter and searching the wire mesh in front of the emitter, the mouse reveals its preference for one sound over another. This model allows comparing the attractiveness of the various features of mouse USVs, in various contexts.

Los USVs registran de una BALB-macho (161 sílabas por 20 sec), así como el ruido de fondo se usaron como la reproducción de sonidos en el experimento representativo se muestra en la Figura 3. En este experimento, se utilizaron 7 ratones B6 hembra a 9 semanas de edad . Para determinar la mejor duración de las pruebas de respuesta femenina para reproducir los sonidos, los parámetros de comportamiento se analizaron por separado para los primeros y últimos cinco min de la época total...

Watch this Scientific Journal Video about Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test at JoVE.com

Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

La determinación de Preferencias ultrasónico Vocalización en ratones utilizando una prueba de reproducción de dos elección

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, ...

determining-ultrasonic-vocalization-preferences-mice-using-two-choice

Research

JoVE Journal

Behavior

11.1K vistas.  Azabu University. Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

Video: Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

The Successive Alleys Test of Anxiety in Mice and Rats

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

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

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

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 ...

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 ...

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

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

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

La dimensión social del estrés: Manipulaciones experimentales de Apoyo Social e Identidad Social en el Trier Social Estrés Prueba

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

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization (USV) Songs

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup retrieval1, juvenile interactions2, opposite and same sex interactions3,4,5, and territorial interactions6. For decades, the USVs have been used by investigators as proxies to study neuropsychiatric and developmental or behavioral disorders7,8,9, and more recently to understand mechanisms and evolution of vocal communication among vertebrates10. Within the sexual interactions, adult male mice produce USV songs, which have some features similar to courtship songs of songbirds11. The use of such multisyllabic repertoires can increase potential flexibility and information they carry, as they can be varied in how elements are organized and recombined, namely syntax. In this protocol a reliable method to elicit USV songs from male mice in various social contexts, such as exposure to fresh female urine, anesthetized animals, and estrus females is described. This includes conditions to induce a large amount of syllables from the mice. We reduce recording of ambient noises with inexpensive sound chambers, and present a quantification method to automatically detect, classify and analyze the USVs. The latter includes evaluation of call-rate, vocal repertoire, acoustic parameters, and syntax. Various approaches and insight on using playbacks to study an animal&#39;s preference for specific song types are described. These methods were used to describe acoustic and syntax changes across different contexts in male mice, and song preferences in female mice.

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization USV Songs

Mice produce a complex multisyllabic repertoire of ultrasonic vocalizations (USVs). These USVs are widely used as readouts for neuropsychiatric disorders. This protocol describes some of the practices we learned and developed to consistently induce, collect, and analyze the acoustic features and syntax of mouse songs.

Eliciando y analizando canciones masculinas de vocalización ultrasónica (USV)

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup ...

Psychophysical Tracking Method to Measure Taste Preferences in Children and Adults

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to adulthood. The method, which is identical for children, adolescents, and adults, is of short duration (&#60; 15 min), does not rely on sustained attention or place demands on memory (which would yield spurious age differences), and minimizes the impact of language development, making this method amenable to the cognitive limitations of pediatric populations. In this whole-mouth tasting method, subjects are asked to taste (without swallowing) pairs of solutions of different sucrose concentrations and to point to the solution they prefer. Each subsequent pair contains the participant&#39;s preceding preferred concentration and an adjacent stimulus concentration. The procedure continues until the subject chooses either a given concentration of sucrose when paired with both a higher and a lower concentration, or the highest or lowest concentration two consecutive times. Subjects are prevented from reaching response criteria on the basis of first or second position bias by the two-series design of the method, which counterbalances the order of solution presentation within each pair between the series (the weaker concentration is presented first in Series 1, second in Series 2). The geometric mean of the two sucrose concentrations chosen in Series 1 and 2 is an estimate of the participant&#39;s most preferred level of sucrose. Sucrose preference as determined with this laboratory-based measure has been shown to be associated with preference for sugars in foods and beverages and with taste receptor genotype, family history of alcoholism, and race/ethnicity, as well as depressive symptomatology among pediatric populations. The method has real-world relevance and has been applied to determine most preferred level of other tastes (e.g., salt), making it a valuable psychophysical tool.

Sweet taste has powerful hedonic appeal among people of all ages, particularly children. Described herein is a reliable and valid method that can be used to determine the level of sweetness most preferred, making it a valuable psychophysical tool for scientists.

Psicofísico Método de Seguimiento para medir las preferencias de sabor en niños y adultos

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

La prueba de roedores psicomotor Vigilancia (rPVT): Un método para evaluar el rendimiento neuroconductual en ratas y ratones

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is fairly simple and can be completed over 3 days: habituation day, training day, and testing day. During training, the mouse is allowed to explore 2 identical objects. On test day, one of the training objects is replaced with a novel object. Because mice have an innate preference for novelty, if the mouse recognizes the familiar object, it will spend most of its time at the novel object. Due to this innate preference, there is no need for positive or negative reinforcement or long training schedules. Additionally, the ORT can also be modified for numerous applications. The retention interval can be shortened to examine short-term memory, or lengthened to probe long-term memory. Pharmacological intervention can be used at various times prior to training, after training, or prior to recall to investigate different phases of learning (i.e., acquisition, early or late consolidation, or recall). Overall, the ORT is a relatively low-stress, efficient test for memory in mice, and is appropriate for the detection of neuropsychological changes following pharmacological, biological, or genetic manipulations.

The object recognition test (ORT) is a simple and efficient assay for evaluating learning and memory in mice. The methodology is described below.

Prueba de reconocimiento de objeto novedoso para la investigación del aprendizaje y la memoria en ratones

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is ...

Reinstatement of Drug-seeking in Mice Using the Conditioned Place Preference Paradigm

The present protocol describes the Conditioned Place Preference (CPP) as a model of relapse in drug addiction. In this model, animals are first trained to acquire a conditioned place preference in a drug-paired compartment, and after the post-conditioning test, they perform several sessions to extinguish the established preference. The CPP permits the evaluation of the conditioned rewarding effects of drugs related to environmental cues. Then, the extinguished CPP can be robustly reinstated by the non-contingent administration of a priming dose of the drug, and by exposure to stressful stimuli. Both methods will be explained here. When the animal reinitiates the behavioral response, a reinstatement of the conditioned reward is considered to have taken place.
The main advantages of this protocol are that it is non-invasive, inexpensive, and simple with good validity criteria. In addition, it allows the study of different environmental manipulations, such as stress or diet, which can modulate relapse into drug seeking behaviors. However, one limitation is that if the researcher aims to explore the motivation and primary reinforcing effects of the drug, it should be complemented with self-administration procedures, as they involve operant responses of animals.

This protocol describes the Conditioned Place Preference (CPP) as a model of relapse. This procedure permits the measurement of relapse in laboratory animals, considering the impact of drug-associated environmental cues as craving and relapse in abstaining addicts is currently the focus of drug-abuse treatment programs.

Reintegro de la búsqueda de drogas en ratones utilizando el paradigma de preferencia condicionada del lugar

The present protocol describes the Conditioned Place Preference (CPP) as a model of relapse in drug addiction. In this model, animals are first trained to ...

Assessment of Social Transmission of Food Preferences Behaviors

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy aging groups. For example, olfactory dysfunction in AD can present as impairment in olfactory recognition, emerging during early stages of the disease and worsening while the disease progresses. The social transmission of food preferences (STFP) task is based on a rudimentary form of communication between rodents concerning distant foods dependent on the transmission of olfactory cues. Healthy wild-type mice would prefer to eat a novel, flavored food that was previously cued by a conspecific, and this food preference would be hampered in transgenic AD mice, such as the APP/PS1 model. Indeed, a strong preference for the cued food in C57Bl6/J mice of 3 months of age was found, and this was reduced in 3 months old transgenic APP/PS1 mice. In summary, STFP task could be a powerful measure to be integrated in present subclinical detection assays of AD.

This paper presents a protocol for the investigation of social transmission of food preference in mice. The advantages and possible applications for this procedure, for instance, in detecting early changes in AD mouse models, are highlighted. To conclude, interpretation of the results in light of critical details are discussed.

Evaluación de la transmisión Social de comportamientos de preferencias de alimentos

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy ...