Studying Socio‐Affective Communication in Rats through Playback of Ultrasonic Vocalizations

Markus Wöhr1, Dominik Seffer1, Rainer K.W. Schwarting1

1 Behavioral Neuroscience, Experimental and Biological Psychology, Philipps‐University of Marburg, Marburg
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 8.35
DOI:  10.1002/cpns.7
Online Posting Date:  April, 2016
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Rats are able to produce ultrasonic vocalizations (USVs). Such USVs are an important component of the rat social behavior repertoire and serve distinct communicative functions as socio‐affective signals. Depending on the emotional valence of the situation, juvenile and adult rats utter (1) aversive 22‐kHz USVs conveying an appeasing and/or alarming function; or (2) appetitive 50‐kHz USVs, which act as social contact calls, amongst others. A 50‐kHz USV radial maze playback paradigm that allows assessment of the behavioral responses displayed by the recipients in a highly standardized manner has been developed. In this newly developed paradigm, a rat is exposed individually to playback of natural 50‐kHz USVs and appropriate acoustic control stimuli using an acoustic presentation system for ultrasound. By this means, it has been consistently shown that 50‐kHz USVs lead to social approach behavior in the recipient, supporting the notion that they serve an affiliative function as social contact calls. © 2016 by John Wiley & Sons, Inc.

Keywords: ultrasonic vocalization; ultrasonic communication; contact call; alarm call; social behavior; social motivation; playback

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Table of Contents

  • Introduction
  • Basic Protocol 1: The 50‐kHz USV Radial Maze Playback Paradigm
  • Support Protocol 1: Sound Calibration
  • Commentary
  • Figures
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Basic Protocol 1: The 50‐kHz USV Radial Maze Playback Paradigm

  • Rats (at least 3 weeks of age; a minimum of 8 rats per experimental condition; 12 rats are recommended)
  • 0.1% acetic acid solution
  • Two PC workstations running Microsoft Windows XP or later
  • Light device for dim red lighting (adjustable, ∼150 cm above maze)
  • Lux meter (e.g., Testo 545 lux meter, Testo)
  • Two ultrasonic loudspeakers for playback of ultrasound (ScanSpeak, Avisoft Bioacoustics)
  • Two ultrasonic microphones for ultrasound recording (UltraSoundGate Condensor Microphone CM16/CMPA, Avisoft Bioacoustics)
  • External sound card with a sampling rate of 192 kHz (e.g., Fire Wire Audio Capture FA‐101, Edirol)
  • Portable ultrasonic power amplifier with a frequency range of 1‐125 kHz (Avisoft Bioacoustics)
  • Video camera (∼150 cm above maze, e.g., Panasonic WV‐BP 330/GE)
  • HDD recorder (e.g., Iomega Screenplay Pro, Iomega International SA)
  • Flat‐screen TV
  • Automated video tracking system (EthoVision XT, Noldus Information Technology)
  • Avisoft UltraSoundGate 416 H USB audio device for connecting microphones to PC workstation (Avisoft Bioacoustics)
  • External hard drive for data storage (e.g., 4 TB My Book, Western Digitals)
  • Computer software:
  • Avisoft SASLab Pro for generating and presenting acoustic stimuli in the ultrasonic range and data analysis (Avisoft Bioacoustics)
  • Avisoft RECORDER USGH for ultrasound recording (Avisoft Bioacoustics)
  • Folding yardstick
  • Radial eight‐arm maze (9.8 × 40.5–cm arms extending radially from a central platform with a diameter of 24 cm, elevated 52 cm above the floor, made of black plastic, and surrounded by a black curtain to minimize visual cues and to reduce sound reflections
  • Sound files of previously recorded 50‐kHz USVs and appropriate controls as stimuli

Support Protocol 1: Sound Calibration

  • G.R.A.S. ultrasonic microphone for recording the reference sound and ultrasound recording (G.R.A.S. 46DP ⅛‐in. LEMO Pressure Standard Microphone Set, G.R.A.S. Sound & Vibration)
  • Avisoft UltraSoundGate power module and input connector for connecting ⅛‐in. microphones directly to the Avisoft UltraSoundGate 416 H USB audio device (UltraSoundGate ¼‐in. Mic Power Module with 7‐pin LEMO 1B input connector; Avisoft Bioacoustics)
  • G.R.A.S. sound calibrator for generating a reference sound (G.R.A.S. Sound Calibrator Type 42AB, G.R.A.S. Sound & Vibration)
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Literature Cited

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Key References
   Brudzynski , 2013. See above.
  A review of the author's and others’ work describing the communicative function of ultrasonic vocalizations and their underlying brain mechanisms.
   Kisko et al., 2015. See above.
  This is a unique study showing, for the first time, that lack of 50‐kHz ultrasonic vocalizations following surgical devocalization results in a reduction of rough‐and‐tumble play in rats.
   Knutson et al., 1998. See above.
  This study is the first presentation of results showing that rats emit 50‐kHz ultrasonic vocalizations during rough‐and‐tumble play.
   Seffer et al., 2015. See above.
  This is an extensive study on the effects of social housing conditions and the first one showing that post‐weaning social isolation leads to a lack of social approach behavior elicited by playback of 50‐kHz ultrasonic vocalizations.
   Willuhn et al., 2014. See above.
  This is the first study using the operant conditioning chamber adaption of the 50‐kHz ultrasonic vocalization playback paradigm for in vivo recordings, showing that 50‐kHz but not 22‐kHz ultrasonic vocalizations elicit dopamine release in the nucleus accumbens.
   Wöhr and Schwarting , 2013. See above.
  A review of the authors’ and others’ work describing the role of ultrasonic vocalizations as socio‐affective signals with important communicative functions and how they can be used in animal models of neuropsychiatric disorders.
Internet Resources
  Avisoft Bioacoustics’ site describes hardware and software available for investigating animal acoustic communication, including equipment to record and analyze rodent ultrasonic vocalizations and to perform playback experiments. It also includes examples of animal sounds and an overview on ultrasonic vocalizations in rodents.
  G.R.A.S. Sound & Vibration's site describes a large set of acoustic equipment, including sound calibration systems that allow calibrating sounds in the ultrasonic range as they are used in playback experiments in rats.‐behavior‐research.
  Noldus Information Technology's site describes several computer systems available for the automated analysis of animal behavior, including behavioral changes elicited in response to playback of ultrasonic vocalizations in rats.
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