Name: determining ultrasonic vocalization preferences in mice using a two-choice playback test
Uploaded: 2015-09-03T15:00:00
Description: Watch this Scientific Journal Video about Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test at JoVE.com

This work was supported by a Grant-in-Aid for JSPS Fellows to A.A.; by Grants-in-Aid for Scientific Research on Innovative Areas for JSPS Fellows (No. 4501 and No. 25132712) to T.K.; and by a research project grant awarded by Azabu University. Figure 2 is reprinted from Kihara, T., Harada, T., &#38; Koshida, N. Wafer-compatible fabrication and characteristics of nanocrystalline silicon thermally induced ultrasound emitters. In: Sensors and Actuators A: Physical, volume 125, Elsevier, p. 426, (2006), with permission from Elsevier.

All procedures were approved by the Ethics Committee of Azabu University. All experiments were carried out in a soundproof chamber.
1. Animal Preparation
<ol>
	<li>Males for Recording
	<ol>
		<li>Obtain sexually mature male&#160;mice experienced in mating.</li>
	</ol>
	</li>
	<li>Female Subjects
	<ol>
		<li>Obtain virgin female mice that have been housed with 2 to 5 littermates per cage (usually 8 - 12 weeks old).</li>
		<li>Obtain vaginal smears daily to determine the phase of the estrous c...

<ol>
	<li>Panksepp, J. B., 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=Affiliative+behavior,+ultrasonic+communication+and+social+reward+are+influenced+by+genetic+variation+in+adolescent+mice.">Affiliative behavior, ultrasonic communication and social reward are influenced by genetic variation in adolescent mice.</a> PLoS One. 2 (4), e351 (2007).</li><li>Scattoni, M. L., Ricceri, L., 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=Unusual+repertoire+of+vocalizations+in+adult+BTBR+T+tf/J+mice+during+three+types+of+social+encounters.">Unusual repertoire of vocalizations in adult BTBR T+tf/J mice during three types of social encounters.</a> Genes Brain Behav. 10 (1), 44-56 (2010).</li><li>Hammerschmidt, K., Radyushkin, K., Ehrenreich, H., Fischer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+structure+and+usage+of+female+and+male+mouse+ultrasonic+vocalizations+reveal+only+minor+differences.">The structure and usage of female and male mouse ultrasonic vocalizations reveal only minor differences.</a> PLoS One. 7 (7), e41133 (2012).</li><li>Merten, S., Hoier, S., Pfeifle, C., Tautz, 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+role+for+ultrasonic+vocalisation+in+social+communication+and+divergence+of+natural+populations+of+the+house+mouse+(Mus+musculus+domesticus).">A role for ultrasonic vocalisation in social communication and divergence of natural populations of the house mouse (Mus musculus domesticus).</a> PLoS One. 9 (5), e97244 (2014).</li><li>Amato, F. R., Moles, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonic+vocalizations+as+an+index+of+social+memory+in+female+mice.">Ultrasonic vocalizations as an index of social memory in female mice.</a> Behav. Neurosci. 115 (4), 834-840 (2001).</li><li>Chabout, J., 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=Adult+male+mice+emit+context-specific+ultrasonic+vocalizations+that+are+modulated+by+prior+isolation+or+group+rearing+environment.">Adult male mice emit context-specific ultrasonic vocalizations that are modulated by prior isolation or group rearing environment.</a> PLoS One. 7 (1), e29401 (2012).</li><li>Ehret, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infant+rodent+ultrasounds&#8211;a+gate+to+the+understanding+of+sound+communication.">Infant rodent ultrasounds&#8211;a gate to the understanding of sound communication.</a> Behav. Genet. 35 (1), 19-29 (2005).</li><li>Holy, T. E., Guo, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonic+songs+of+male+mice.">Ultrasonic songs of male mice.</a> PLoS Biol. 3 (1), e386 (2005).</li><li>Portfors, C. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Types+and+functions+of+ultrasonic+vocalizations+in+laboratory+rats+and+mice.">Types and functions of ultrasonic vocalizations in laboratory rats and mice.</a> J. Am. Assoc. Lab. Anim. Sci. 46 (1), 28-34 (2007).</li><li>Holfoth, D. P., Neilans, E. G., Dent, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discrimination+of+partial+from+whole+ultrasonic+vocalizations+using+a+go/no-go+task+in+mice.">Discrimination of partial from whole ultrasonic vocalizations using a go/no-go task in mice.</a> J. Acoust. Soc. Am. 136 (6), 3401 (2014).</li><li>Neilans, E. G., Holfoth, D. P., Radziwon, K. E., Portfors, C. V., Dent, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discrimination+of+ultrasonic+vocalizations+by+CBA/CaJ+mice+(Mus+musculus)+is+related+to+spectrotemporal+dissimilarity+of+vocalizations.">Discrimination of ultrasonic vocalizations by CBA/CaJ mice (Mus musculus) is related to spectrotemporal dissimilarity of vocalizations.</a> PLoS One. 9 (1), e85405 (2014).</li><li>Hammerschmidt, K., Radyushkin, K., Ehrenreich, H., Fischer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Female+mice+respond+to+male+ultrasonic+&#8216;songs&#8217;+with+approach+behaviour.">Female mice respond to male ultrasonic &#8216;songs&#8217; with approach behaviour.</a> Biol. Lett. 5 (5), 589-592 (2009).</li><li>Musolf, K., Hoffmann, F., Penn, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonic+courtship+vocalizations+in+wild+house+mice,+Mus+musculus+musculus.">Ultrasonic courtship vocalizations in wild house mice, Mus musculus musculus.</a> Anim. Behav. 79 (3), 757-764 (2010).</li><li>Asaba, 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=Developmental+social+environment+imprints+female+preference+for+male+song+in+iice.">Developmental social environment imprints female preference for male song in iice.</a> PloS one. 9 (2), e87186 (2014).</li><li>Uematsu, 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=Maternal+approaches+to+pup+ultrasonic+vocalizations+produced+by+a+nanocrystalline+silicon+thermo-acoustic+emitter.">Maternal approaches to pup ultrasonic vocalizations produced by a nanocrystalline silicon thermo-acoustic emitter.</a> Brain Res. 1163, 91-99 (2007).</li><li>Okabe, S., 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=The+effects+of+social+experience+and+gonadal+hormones+on+retrieving+behavior+of+mice+and+their+responses+to+pup+ultrasonic+vocalizations.">The effects of social experience and gonadal hormones on retrieving behavior of mice and their responses to pup ultrasonic vocalizations.</a> Zoolog. Sci. 27 (10), 790-795 (2010).</li><li>Okabe, S., 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=Pup+odor+and+ultrasonic+vocalizations+synergistically+stimulate+maternal+attention+in+mice.">Pup odor and ultrasonic vocalizations synergistically stimulate maternal attention in mice.</a> Behav. Neurosci. 127 (3), 432-438 (2013).</li><li>Shinoda, H., Nakajima, T., Ueno, K., Koshida, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermally+induced+ultrasonic+emission+from+porous+silicon.">Thermally induced ultrasonic emission from porous silicon.</a> Nature. 400 (6747), 853-855 (1999).</li><li>Kihara, T., Harada, T., Koshida, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wafer-compatible+fabrication+and+characteristics+of+nanocrystalline+silicon+thermally+induced+ultrasound+emitters.">Wafer-compatible fabrication and characteristics of nanocrystalline silicon thermally induced ultrasound emitters.</a> Sensor. Actuat. A-Phys. 125 (2), 422-428 (2006).</li><li>Kikusui, T., 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=Cross+fostering+experiments+suggest+that+mice+songs+are+innate.">Cross fostering experiments suggest that mice songs are innate.</a> PloS One. 6 (3), e17721 (2011).</li><li>McLean, A. C., Valenzuela, N., Fai, S., Bennett, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performing+Vaginal+Lavage,+Crystal+Violet+Staining,+and+Vaginal+Cytological+Evaluation+for+Mouse+Estrous+Cycle+Staging+Identification.">Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification.</a> J. Vis. Exp. (67), e4389 (2012).</li><li>Nelson, J. F., Felicio, L. S., Randall, P. K., Sims, C., Finch, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+longitudinal+study+of+estrous+cyclicity+in+aging+C57BL/6J+mice:+I.+Cycle+frequency,+length+and+vaginal+cytology.">A longitudinal study of estrous cyclicity in aging C57BL/6J mice: I. Cycle frequency, length and vaginal cytology.</a> Biol. Reprod. 27 (2), 327-339 (1982).</li><li>Tomihara, K., 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=Effect+of+ER-beta+gene+disruption+on+estrogenic+regulation+of+anxiety+in+female+mice.">Effect of ER-beta gene disruption on estrogenic regulation of anxiety in female mice.</a> Physiol. Behav. 96 (2), 300-306 (2009).</li><li>Zheng, Q. Y., Johnson, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hearing+loss+associated+with+the+modifier+of+deaf+waddler+(mdfw)+locus+corresponds+with+age-related+hearing+loss+in+12+inbred+strains+of+mice.">Hearing loss associated with the modifier of deaf waddler (mdfw) locus corresponds with age-related hearing loss in 12 inbred strains of mice.</a> Hear. Res. 154 (1-2), 45-53 (2001).</li><li>Haga, S., 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=The+male+mouse+pheromone+ESP1+enhances+female+sexual+receptive+behaviour+through+a+specific+vomeronasal+receptor.">The male mouse pheromone ESP1 enhances female sexual receptive behaviour through a specific vomeronasal receptor.</a> Nature. 466 (7302), 118-122 (2010).</li><li>Grimsley, J. M., Monaghan, J. J., Wenstrup, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+social+vocalizations+in+mice.">Development of social vocalizations in mice.</a> PLoS One. 6 (3), e17460 (2011).</li><li>Sugimoto, H., 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=A+role+for+strain+differences+in+waveforms+of+ultrasonic+vocalizations+during+male-female+interaction.">A role for strain differences in waveforms of ultrasonic vocalizations during male-female interaction.</a> PLoS ONE. 6 (7), (2011).</li><li>Hanson, J. L., Hurley, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Female+presence+and+estrous+state+influence+mouse+ultrasonic+courtship+vocalizations.">Female presence and estrous state influence mouse ultrasonic courtship vocalizations.</a> PLoS One. 7 (7), e40782 (2012).</li><li>Wang, H., Liang, S., Burgdorf, J., Wess, J., Yeomans, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonic+vocalizations+induced+by+sex+and+amphetamine+in+M2,+M4,+M5+muscarinic+and+D2+dopamine+receptor+knockout+mice.">Ultrasonic vocalizations induced by sex and amphetamine in M2, M4, M5 muscarinic and D2 dopamine receptor knockout mice.</a> PLoS One. 3 (4), (2008).</li><li>Moles, A., Kieffer, B. L., D'Amato, F. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deficit+in+attachment+behavior+in+mice+lacking+the+mu-opioid+receptor+gene.">Deficit in attachment behavior in mice lacking the mu-opioid receptor gene.</a> Science. 304 (5679), 1983-1986 (2004).</li><li>Hiramoto, T., 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=Tbx1:+identification+of+a+22q11.2+gene+as+a+risk+factor+for+autism+spectrum+disorder+in+a+mouse.">Tbx1: identification of a 22q11.2 gene as a risk factor for autism spectrum disorder in a mouse.</a> Hum. Mol. Genet. 20 (24), 4775-4785 (2011).</li><li>Ey, 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=Absence+of+deficits+in+social+behaviors+and+ultrasonic+vocalizations+in+later+generations+of+mice+lacking+neuroligin4.">Absence of deficits in social behaviors and ultrasonic vocalizations in later generations of mice lacking neuroligin4.</a> Genes Brain Behav. , (2012).</li><li>Roy, S., Watkins, N., Heck, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+analysis+of+ultrasonic+vocalizations+in+a+mouse+model+of+fragile+X+syndrome+reveals+limited,+call+type+specific+deficits.">Comprehensive analysis of ultrasonic vocalizations in a mouse model of fragile X syndrome reveals limited, call type specific deficits.</a> PLoS One. 7 (9), e44816 (2012).</li></ol>

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...

Many animals use vocalizations for intraspecific communication. In the mouse Mus musculus, one important type of communication signal is ultrasonic vocalizations (USVs), which have frequencies higher than 20 kHz. USVs emitted by mice are considered a component of social recognition in male-female1-4, female-female1, 5, and male-male1, 6 interactions. USVs are also emitted by pups when they are isolated from their mother, which increases her pup-retrieving behavior, and therefore pup survival 7. Although many reports have analyzed and categorized mouse USVs8,9, the behavioral responses and neural mechanisms of the receiving animal have been less documented10, 11. The latter is necessary to clarify the biological significance of the various characteristics of USVs. To reveal these mechanisms, the playback experiment is an efficient method. Recent playback studies have revealed that female mice are attracted to USVs12, and that they prefer USVs from males that are different from their parents13, 14.
This article explains the playback test used to evaluate USV preference in mice. A two-choice test box was developed in which two different USVs can be played back simultaneously in two compartments of a test enclosure, as shown in Figure 1. This type of test box prevents sound contamination by dividing the test area in three sub-chambers, using lead walls. The ultrasound emitters are located outside each room. In the wall between the rooms and ultrasound emitters are holes covered with wire mesh. Mice can move freely in the three rooms, and show a &#8220;searching the mesh&#8221; behavior, as if to respond to USVs played back by the ultrasound emitters. In this test, mice stay for periods of different duration close to one sound emitter or the other. These parameters can be logged to obtain a sensitive measure of sound preference.
To play the USVs back, nanocrystalline silicon thermo-acoustic emitters (i.e., &#8220;nc-Si emitter&#8221;) were used as in previous studies15-17. These devices are composed of a thin-film heater electrode, a nano-porous silicon layer, and a single-crystalline silicon wafer. The digital sound file is converted to an analog signal and then passed through the heater electrode. The device converts the resulting voltage-dependent thermal signals into significant sound pressure with low distortion. This device is unique in that, unlike common sound generators that depend on mechanical vibrations, it can reproduce sound without the need of a diaphragm. The emitter exhibits a flat sound pressure level at frequencies from 20 to 160 kHz (Figure 2), and can reproduce digitally recorded murine USVs very accurately in terms of duration, frequency, and sound pressure level15, 18, 19.
In a representative experiment shown in Figure 3, C57BL/6 (B6) females were allowed to choose between BALB/c (BALB) male USVs and background noise. In addition, Figure 4 shows the choice of B6 and BALB females between simultaneous USV playbacks from a BALB and a B6 male, as reported in a previous study14. The characteristics of male USVs differ between B6 and BALB strains20. As shown by these results, the attractiveness of USVs can be assessed with the present protocol, in which sounds are recorded from a live individual, acoustically analyzed, and played back to other individuals.

<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.

The USVs recorded from one BALB-male (161 syllables per 20 sec), as well as background noise were used as playback sounds in the representative experiment shown in Figure 3. In this experiment, 7 female B6 mice were used at 9 weeks of age. To determine the best duration of testing of female response to playback sounds, the behavioral parameters were analyzed separately for the first and last five min of the total 10 min test time.
First, there was no significant difference in ...

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.

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

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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 ...

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 ...

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

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 ...

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 ...

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 ...

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 ...