Diese Arbeit wurde durch ein Grant-in-Aid für JSPS-Stipendiaten auf AA; von Grants-in-Aid for Scientific Research für innovative Bereiche für JSPS-Stipendiaten (No. 4501 und Nr 25132712) zu TK; und von einem Forschungsprojekt Zuschuss von Azabu University. Abbildung 2 verliehen wird aus Kihara, T., Harada, T., &amp; Koshida, N. Wafer-kompatible Herstellung und Eigenschaften der nanokristallinen Silizium thermisch induzierte Ultraschallsender nachgedruckt. In: Sensors and Actuators A: Physical, Band 125, Elsevier, p. 426, (2006), mit Genehmigung von Elsevier.

<html> Alle Verfahren wurden von der Ethikkommission der Azabu Universität genehmigt. Alle Versuche wurden in einer schalldichten Kammer durchgeführt wird. 1. Vorbereitung der Tiere <ol><li> Männliche Person für die Aufnahme <ol><li> Erhalten geschlechtsreifen männlichen Mäusen in Paarungs erlebt. </li></ol></li><li> Probandinnen <ol><li> Erhalten jungfräulichen weiblichen Mäusen, die mit 2 bis 5 Wurf pro Käfig gehalten worden sind (in der Regel 8 bis 12 Wochen alt). </li><li> Erhalten Vaginalabstrichen täglich, um die Phase des Östruszyklus vor der Prüfung zu bestimmen, nach McLean 21. Es sollte darauf geachtet, um die vaginale Stimulation zu minimieren, um Scheinträchtigkeit vermeiden. </li><li> Färben die Ausstriche mit Giemsa und bestimmen Phase des Östruszyklus basierend auf dem Vorhandensein oder der Abwesenheit von Leukozyten, verhornten Epithelzellen und kernhaltige Epithelzellen, nach Nelson 22. </li></ol></li></ol> 2. Geräte (Abbildung 1) _content &quot;&gt; <img alt="Abbildung 1" src="/files/ftp_upload/53074/53074fig1.jpg" /> Abbildung 1. Schematische Darstellung des Zwei-Choice-Test-Box und Geräte. Die Mäuse können durch die kleinen Toren zwischen ihnen und der neutralen Zone zugreifen Räume A und B. Die zwei Auswahlbox und zwei Ultraschallsender sind auf dem Boden von einer schalldichten Kammer platziert. Das Mikrofon ist in der schalldichten Kammer suspendiert. Abkürzung:. PC, PC <a href="https://www.jove.com/files/ftp_upload/53074/53074fig1large.jpg" target="_blank">Bitte klicken Sie hier, um eine größere Version dieser Figur zu sehen.</a> <ol><li> Für Recording <ol><li> Schließen Sie ein Ultraschallkondensatormikrofon an einen PC über ein USB-A / D-Wandler. </li><li> Stellen Sie den Verstärkungspegel des A / D-Wandler auf &quot;7&quot;. </li><li> Nehmen akustischen Daten in 16-Bit-Format-Dateien mit einer Abtastrate von 400 kHz mittels Audio-Capture-Software. </li></ol></li><li> Für die Wiedergabe <ol><li> Schließen Sie einen D / A-Wandler mit dem PC übereine USB-Schnittstelle. </li><li> Verbindung von zwei Verstärkern in den D / A-Wandler durch ein Dämpfungsglied. (Letzteres Einrichtung dient, um eine Überlastung zu verhindern.) </li><li> Schließen Sie ein Ultraschallsender an jeden Verstärker. </li></ol></li></ol> 3. USV Recording <ol><li> Legen Sie eine männliche Maus in einem kleinen (17 x 10 x 11 cm) und mittlere (21 x 14 x 13 cm) Käfig mit einer 6 cm-Durchmesser-Loch in der Seitenwand und dem Loch mit einem 0,5 cm Drahtgitter abgedeckt. Bei der Verwendung eines mittelgroßen Käfig, konstruieren Sie eine Partition auf der Mitte des Käfigs, um den Bereich zu reduzieren. </li><li> Sie Bettwäsche in den Käfig zu verwenden nicht Rascheln Geräusche, die die Aufnahmen kontaminieren könnten. </li><li> Legen Sie ein Mikrofon neben dem Mesh. </li><li> Beginnen Ultraschall vocalization Überwachung mit dem Aufnahme-Setup in Schritt 2.1 dargestellt. </li><li> Legen Sie eine geschlechtsreifen weiblichen Maus entweder Metöstrus oder Diöstrus in den Käfig mit dem männlichen Maus. </li><li> Nehmen USVs für 3-5 min. </li></ol> 4. Test Box <ol><li> Erstellen Sie das Testfeld mit Acrylplatte (Abbildung 1). Hinweis: Das Testfeld (35 x 20 und x 20 cm hoch) ist in drei Abteilungen unterteilt: eine neutrale Zone (15 x 20 cm), und die Zimmer A und B (20 x 10 cm, jeweils). Zimmer A und B jeweils eine 4 cm Durchmesser Loch, das mit einem 0,5 cm Drahtgewebe abgedeckt ist. Die Löcher und Mesh sind an jedem Ende der Zimmer A und B. Achten Tore (5 x 5 cm jeweils) zwischen der neutralen Zone und Zimmer A und B befindet, </li><li> Zu verhindern Tonverlust isolieren jedes Ultraschallsender, mit Ausnahme der Seite, die Gitterwand, mit Gummiplatten, Dichtung in die Schachtel um den Umfang jeder Masche. </li><li> Platz Plomben auf beiden Seiten der Mauer zwischen den Räumen A und B für den Schallschutz. </li></ol> 5. Vorbereitung für Soundwiedergabe <ol><li> Schaffung Wiedergabe Sounds <ol><li> Nehmen Sie ein 20 sec Segment der USV aus der aufgezeichneten Datei mit digitalen Audio-Editing-Software. </li><li> Wählen und erfassen Hintergrund keineise (0,35 sec) aus dem 20 sec USV-Segment-Datei. Erstellen Sie eine 20 Sekunden Hintergrundrauschen-Datei durch Wiederholen des Rauschsegment. </li><li> Um die beiden Audio-Dateien gleichzeitig wiedergeben, exportieren Sie sie als eine einzelne Stereo-Sound-Datei mit einer Datei für den &quot;linken Ohr&quot; und die andere Datei für die &quot;rechte Ohr.&quot; </li><li> Filtern Sie die Sound-Dateien, wobei als Hochpassfilter mit einem Cut-off von 40 kHz. </li><li> Verringern Sie Geräusche mit dem Rauschunterdrückungswerkzeug im Post-Processing-Software. Wählen und erfassen ein Segment der Datei, die keine USVs wie die Rauschunterdrückungs Profil enthält, und führen Sie Rauschreduktion mit Level 40. </li></ol></li><li> Kalibrierung der Wiedergabe Sounds <ol><li> Sicherzustellen, dass der Raum zwischen Ultraschallsender und Mikrofon 10 cm. </li><li> Schalldruckpegel von den Ultraschallsender zu kalibrieren, führen Ultraschallüberwachung unter Verwendung der in Schritt 2.1 beschriebenen Konfiguration. Messen Sie den Schalldruckpegel von dem Mikrofon in Dezibel (dB). </li><li> Using der Dämpfer und Verstärker, die Lautstärke der von den Ultraschallsender auf den gleichen Schalldruck wie die männlichen USVs in Schritt 3 aufgezeichnet erzeugt USVs. </li><li> Bei der Verwendung der zwei Ultraschallsender und eine Audio-Datei von USVs und Hintergrundrauschen besteht, zu bestätigen, dass die beiden Ultraschallsender weisen die gleiche Schalldruckpegel unter Verwendung einer Eich Sound (zB ein 75-kHz reiner Ton) vor der Durchführung des Verhaltenstest. </li><li> Beim Wiedergeben einer Audiodatei von zwei Strömen von USVs besteht, passen die beiden erzeugten Schalldruckpegel auf den gleichen Pegel, vor dem Test. </li></ol></li></ol> 6. Zwei-Choice-Test <ol><li> Legen Sie die Ultraschallsender hinter den Maschen des Testfeld. </li><li> Schließen Sie die Türen zu Räumen A und B mit einer Acrylplatte und gewöhnen die weibliche unterliegen der neutralen Zone für 30 min. Decken Sie die Box mit Acrylbrett, um zu vermeiden, dass die Maus entweichen kann. </li><li> Starten Sie Videoaufzeichnung mit einer CCÜber der Box montiert D-Kamera. Die Kamera (in 1 nicht dargestellt) deckt den Bereich aller drei Abteilen. </li><li> Entfernen Sie das Tor und Cover-Board und lassen Sie die weiblichen frei das Testfeld, darunter die Klangzonen zu erkunden. </li><li> Sobald das Weibchen hat beide Netze untersucht und hat sich in die Neutralzone zurückgeführt, spielen Sie eine 20 sec Tondatei wiederholt für 5 oder 10 min. </li><li> Zuführen Verhaltensüberwachung für 5 oder 10 min. </li><li> Um unerwünschte olfaktorischen Signale aus dem vorherigen Thema hinterlegt reduzieren, reinigen Sie die Testkäfige zwischen den Tests mit 70% Ethanol. </li><li> Schalten Sie den Ort der Ultraschallsender A und B nach dem Zufallsprinzip inhärenten Seiten Bias-Effekt in Richtung Zimmer A oder B zu vermeiden </li></ol> 7. Statistische Analysen <ol><li> Verwenden von Verhaltens Event-Scoring-Software analysiert die folgenden Parameter: Gesamtzahl der Einträge in jedem Zimmer; Gesamtdauer des Aufenthalts in jedem mittleren Zone; Gesamtdauer des Aufenthalts in jedem Ton Zone; undGesamtdauer der Suche nach jeder Masche vor der Ultraschallsender. 1 veranschaulicht, was mit &quot;Mittelzone&quot; und meinte &quot;Sound-Zone.&quot; Hinweis: In den Fällen, wo die Maus bleibt auf der Trennlinie zwischen den beiden Zonen (Abbildung 1, gestrichelte Linie) entscheiden, die Position durch die Richtung des Kopfes. Wenn der Kopf steht vor der Netzseiten, es als ein Aufenthalt in der Klangzone erzielt wird. </li><li> Für jede Verhaltensparameter, zu vergleichen, Antworten für Räume A und B mit einem Wilcoxon-Test, oder gepaarten t -Tests mit einem Signifikanzniveau von 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...

Viele Tiere nutzen Lautäußerungen für intraspezifische Kommunikation. In der Maus Mus musculus, ist eine wichtige Art der Kommunikationssignal Ultraschall vocalizations (USVs), die höhere Frequenzen als 20 kHz aufweisen. USVs von Mäusen emittiert werden als eine Komponente der sozialen Anerkennung in Mann-Frau-1-4, Buchse-Buchse 1, 5, und Stecker-Stecker 1, 6-Interaktionen. USVs werden auch von Jungtieren emittiert, wenn sie von ihrer Mutter, die ihrem Welpen-Abrufen Verhalten erhöht isoliert und daher Pup Überleben 7. Obwohl viele Berichte wurden analysiert und kategorisiert Maus USVs 8, 9, die Verhaltensreaktionen und neuronalen Mechanismen des Aufnahme Tier wurden weniger gut dokumentierte 10, 11. Letzteres ist notwendig, um die biologische Bedeutung der verschiedenen Merkmale der USVs verdeutlichen. Diese Mechanismen offenbaren, ist die Wiedergabeexperiment ein effizientes Verfahren. Aktuelle Wiedergabe Studien haben ergeben, dass weiblicheMäuse werden zur USVs 12 angezogen wird, und dass sie USVs von Männchen, die von ihren Eltern 13, 14 unterschiedlich sind bevorzugen. Dieser Artikel erklärt, die Wiedergabetest verwendet werden, um USV Präferenz bei Mäusen zu bewerten. Ein Zwei-Choice-Test Feld entwickelt, bei denen zwei unterschiedliche USVs gleichzeitig wieder in zwei Kammern eines Testgehäuse, wie in 1 gezeigt, wiedergegeben werden. Diese Art der Testbox verhindert störende Verunreinigungen durch Dividieren der Testbereich in drei Teilkammern , unter Verwendung von Blei Wänden. Die Ultraschallsender werden vor jedem Zimmer befindet. In der Wand zwischen den Zimmern und Ultraschallsender sind Löcher mit Drahtgeflecht bedeckt. Mäuse können frei in den drei Räumen bewegen und zeigen eine &quot;Suchen Sie das Netz&quot; Verhalten, wie um zu USVs von den Ultraschallsender abgespielt reagieren. Bei diesem Test Mäuse bleiben für Zeiträume von unterschiedlicher Dauer in der Nähe einer Schallsender oder das andere. Diese Parameter können angemeldet sein, um einen empfindlichen measu erhalten<html> re Schall Präferenz. Um die USVs wiedergeben, nanokristallinen Silizium thermo-akustischen Emitter (dh &quot;nc-Si-Emitter&quot;) wurden als in früheren Studien 15-17 verwendet. Diese Geräte werden von einem Dünnschicht Heizelektrode eine nanoporöse Siliziumschicht, und einem einkristallinen Silicium-Wafer besteht. Die digitale Audiodatei in ein analoges Signal umgewandelt und dann durch den Heizer Elektrode geleitet. Das Gerät wandelt die resultierende spannungsabhängigen thermischen Signale in signifikanten Schalldruck bei geringer Verzerrung. Diese Vorrichtung ist dadurch einzigartig, dass, im Gegensatz zu gemeinsamen Schallgeneratoren, die von mechanischen Schwingungen abhängt, kann Ton, ohne die Notwendigkeit einer Membran, zu reproduzieren. Der Emitter weist eine flache Schalldruckpegel bei Frequenzen von 20 bis 160 kHz (Abbildung 2), und kann digital aufgezeichnet murine USVs sehr genau in Bezug auf die Dauer, Häufigkeit und Schalldruckpegel 15, 18, ​​19 zu reproduzieren. ve_content &quot;&gt; In einer in Figur 3 gezeigten repräsentativen Experiment wurden C57BL / 6 (B6) Weibchen darf zwischen BALB / c (BALB) männliche USVs und Hintergrundrauschen zu wählen. Ferner zeigt 4 die Wahl B6 und BALB Frauen zwischen gleichzeitige USV Wiedergaben von einer BALB und B6 male, wie in einer früheren Studie berichteten 14. Die Eigenschaften der männlichen USVs unterscheiden zwischen B6 und BALB Stämme 20 dar. Wie gezeigt durch diesen Ergebnissen kann die Attraktivität USVs der vorliegenden Protokoll bewertet werden, in welcher Töne aus einem Live einzelnen aufgezeichnet, akustisch analysiert, und zurück zu anderen Personen gespielt.

<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.

Die USVs von einer BALB-männlich (161 Silben pro 20 sec), als auch Hintergrundgeräusche aufgenommen wurden, wie die Wiedergabe verwendet klingt in der in 3 gezeigten repräsentativen Experiment. In diesem Experiment wurden 7 weiblichen B6-Mäusen am Alter von 9 Wochen verwendet . Zur Ermittlung der besten Testdauer von weiblichen Reaktion zur Wiedergabe von Ton wurden die Verhaltensparameter getrennt für die ersten und letzten fünf Minuten der insgesamt 10 min Testzeit analysiert. <p class="jove...

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.

Bestimmen Ultraschall Vocalization Preferences in Mäuse mit einer Zwei-Choice-Test-Playback

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

11.0K Aufrufe.  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.

Serie: 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.

Die sukzessive Gassen Test von Angst bei Mäusen und Ratten

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.  ...

Forschung

Verhalten

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.

Kontextuelle und Cued Angst Anlage Test mit einer Video-Analyse-System bei Mäusen

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.

Der modifizierte Loch Board - Mess Verhalten, Kognition und Sozialinteraktion an Mäusen und Ratten

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).

Die soziale Dimension der Stress: experimentelle Manipulationen an sozialer Unterstützung und Sozial Identität in der Trier Social Belastungsprobe

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.

Auslösen und Analysieren von männlichen Maus Ultraschall-Vokalisierung (USV) Lieder

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.

Psycho-physischen Tracking-Methode zur Geschmackspräferenzen bei Kindern messen und Erwachsene

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.

Das Nagetier Psychomotorik Vigilance Test (rPVT): Eine Methode zur Beurteilung der Neuropsychologische Leistung bei Ratten und Mäusen

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.

Neuartige Objekt Anerkennung Test zur Untersuchung von lernen und Gedächtnis bei Mäusen

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.

Wiedereinführung der Drug-seeking bei Mäusen mit dem klimatisierten Ort Präferenz Paradigma

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.

Bewertung der sozialen Übertragung von Essen Vorlieben Verhaltensweisen

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