Ultrasonic Vocalizations in Rat and Mouse Pups

Myron A. Hofer1, Harry N. Shair1, Susan A. Brunelli1

1 Columbia University and Sackler Institute of Developmental Psychobiology and New York State Psychiatric Institute, New York, New York
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 8.14
DOI:  10.1002/0471142301.ns0814s17
Online Posting Date:  February, 2002
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The ultrasonic vocalization (USV), or isolation calling response, of infant rats and mice has been studied as a measure of the intensity of an aversive affective state and as an early communicative behavior between pup and mother. The four protocols described in this unit are for the basic isolation testing procedure, and for elicitation of the contact quieting response to littermates and/or dam, the potentiation of isolation calling response by a prior brief maternal interaction and the predatorā€induced suppression of USV by the scent of an unfamiliar male. These procedures for the elicitation of USV, and for its regulation by different kinds of social interaction, provides the basis for experimental research on the early development of emotion and communication in an animal model system.

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Table of Contents

  • Basic Protocol 1: The Isolation Test
  • Alternate Protocol 1: Contact Quieting
  • Alternate Protocol 2: Maternal Potentiation
  • Alternate Protocol 3: Predator‐Induced USV Suppression
  • Commentary
  • Figures
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Basic Protocol 1: The Isolation Test

  • Dam and litter of 2‐ to 17‐day old rat pups or 2‐ to 14‐day old mouse pups
  • Heating pad regulated by feedback from a thermosensor
  • Fine flexible thermistor (Kent Scientific Corp.)
  • Ink marker, unscented
  • Two or more identical test chambers (see above)
  • Ultrasound detector (e.g., Pettersson Elektronik AB or Ultrasound Advice; see above)

Alternate Protocol 1: Contact Quieting

  • Noninhalant anesthetic (e.g., urethane)

Alternate Protocol 2: Maternal Potentiation

  • For materials, see protocol 1.

Alternate Protocol 3: Predator‐Induced USV Suppression

  • Adult male, 150 to 200 days old, housed in a separate room from the dams and their litter to avoid any familiarization with its scent
  • Urethane
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Literature Cited

Literature Cited
   Allin, J.T. and Banks, E.M. 1970. Effects of temperature on ultrasound production by infant albino rats. Dev. Psychobiol. 4:149‐156.
   Amsel, A., Radek, C.C., Graham, M., and Letz, R. 1977. Ultrasound emission in infant rats as an indicant of arousal during appetitive learning and extinction. Science 197:786‐788.
   Benton, D. and Nastiti, K. 1988. The influence of psychotropic drugs on the ultrasonic calling of mouse pups. Psychopharmacology 95:99‐102.
   Blumberg, M.S. and Alberts, J.R. 1990. Ultrasonic vocalizations by rat pups in the cold: An acoustic by‐product of laryngeal braking? Behav. Neurosci. 104:808‐817.
   Blumberg, M.S. and Sokoloff, G. 1998. Thermoregulatory competence and behavioral expression in the young of altricial species: Revisited. Dev. Psychobiol. 33:107‐123.
   Brunelli, S.A. and Hofer, M.A. 1996. Development of ultrasonic vocalization responses in genetically heterogeneous National Institute of Health (N:NIH) rats. Part II. Associations among variables and behaviors. Dev. Psychobiol . 29:517‐528.
   Brunelli, S.A. and Hofer, M.A. 2001. Selective breeding for an infantile phenotype (isolation calling): A window on developmental processes. In Handbook of Behavioral Neurobiology (E. Blass, ed.). Plenum Press, New York.
   Brunelli, S.A., Vinocur, D.D., Soo Hoo, D., and Hofer, M.A. 1997. Five generations of selective breeding for ultrasonic vocalization (USV) responses in N:NIH strain rats. Dev. Psychobiol 31:255‐265.
   Brunelli, S.A., Masmela, J.R., Shair, H.N., and Hofer, M.A. 1998. Effects of biparental rearing on ultrasonic vocalization (USV) responses of rat pups (Rattus norvegicus). J. Comp. Psychol. 112:331‐343.
   Brunner, D., Buhot, M.C., Hen, R., and Hofer, M.A. 1999. Anxiety, motor activation and maternal‐infant interactions in 5HT1B knockout mice. Behav. Neurosci. 113:587‐601.
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   Ehret, G. and Koch, M. 1989. Ultrasound‐induced parental behaviour in house mice is controlled by female sex hormones and parental experience. Ethology 80:81‐93.
   Haack, B., Markl, H., and Ehret, G. 1983. Sound communication between parents and offspring. In The Auditory Psychobiology of the Mouse (J. Willott, ed.) pp. 57‐97. Charles C. Thomas Publisher, Ltd Springfield, Ill.
   Hahn, M.E., Karkowski, L., Weinreb, L., Henry, A., Schanz, N., and Hahn, E.M. 1998. Developmental patterns in the calls of mice 2‐12 days of age. Behav. Genet. 28:315‐325.
   Hofer, M.A. 1996. Multiple regulators of ultrasonic vocalization in the infant rat. Psychoneuroendocrinology 21:203‐217.
   Hofer, M.A. and Shair, H.N. 1980. Sensory processes in the control of isolation‐induced ultrasonic vocalization by two week‐old rats. J. Comp. Physiol. Psychol. 94:271‐279.
   Hofer, M.A. and Shair, H.N. 1991. Independence of ultrasonic vocalization and thermogenic responses in infant rats. Behav.Neurosci. 105:41‐48.
   Hofer, M.A. and Shair, H.N. 1992. Ultrasonic vocalization by rat pups during recovery from deep hypothermia. Dev. Psychobiol 25:511‐528.
   Hofer, M.A. and Shair, H.N. 1993. Ultrasonic vocalization, laryngeal braking and thermoregulation in rat pups: A reappraisal. Behav. Neurosci. 107:354‐362.
   Hofer, M.A., Brunelli, S.A., and Shair, H.N. 1993. The effects of 24 hr maternal separation and of litter size reduction on the isolation distress response of 12 day old rat pups. Dev. Psychobiol 26:483‐497.
   Hofer, M.A., Brunelli, S.A., Masmela, J.R., and Shair, H.N. 1996. Maternal interactions prior to separation potentiate isolation‐induced calling in rat pups. Behav. Neurosci. 110:1158‐1167.
   Hofer, M.A., Masmela, J.R., Brunelli, S.A., and Shair, H.N. 1998. The ontogeny of maternal potentiation of the infant rats' isolation call. Dev. Psychobiol. 33:189‐201.
   Naito, H. and Tonoue, T. 1987. Sex differences in ultrasound distress call by rat pups. Behav. Brain Res. 25:13‐21.
   Newman, J.D. 1991. Vocal manifestations of anxiety and their pharmacological control. In Psychopharmacology of Anxiolytics and Antidepressants (S.E. File, ed.) pp. 251‐260. Pergamon Press,Inc., New York.
   Nitschke, W., Bell, N.J., and Zachman, T. 1972. Distress vocalizations of young in three inbred strains of mice. Dev. Psychobiol. 5:363‐370.
   Roberts, L.H. 1975. Evidence for the laryngeal source of ultrasonic and audible cries of rodents. J. Zool. (London) 175:243‐257.
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   Shair, H.N., Masmela, J.R., and Hofer, M.A. 1999. The influence of olfaction on potentiation and inhibition of ultrasonic vocalization of rat pups. Physiol. Behav. 65:769‐772.
   Smotherman, W.P., Bell, R.W., Hershberger, W.A., and Coover, G.D. 1978. Orientation to rat pup cues: Effects of maternal experiential history. Anim. Behav. 26:265‐273.
   Takahashi, L.K. 1992. Developmental expression of defensive responses during exposure to conspecific adults in preweanling rats (Rattus norvegicus). J. Comp. Psychol. 106:69‐77.
   Zimmerberg, B., Brunelli, S.A., and Hofer, M.A. 1994. Reduction of rat pup ultrasonic vocalizations by the neuroactive steroid, allopregnanolone. Pharm. Biochem. Behav. 47:735‐738.
Key References
   Hofer, 1996. See above.
  A review of the authors' and others' work describing the role of USV in the behavioral regulatory system involving the mother‐pup interaction, the external sensory regulators, and internal physiological and brain systems controlling infant USV.
   Newman, 1991. See above.
  An extensive review of the pharmacological evidence for vocalization as a measure of anxiety in adults as well as young in a variety of species.
   Sales, G. and Pye, D. 1974. Ultrasonic Communication by Animals. Chapman and Hall, London; John Wiley & Sons, New York.
  The classic work on the physical nature of ultrasound, methods of detection and roles in communication in species from insects to mammals and in echolocation by bats.
Internet Resources
  These sites give descriptions of acoustic analytic principles as well as specifications and prices of ultrasonic detectors, microphones, and loudspeakers for playback. Keith Maries at Ultrasound Advice and Lars Pettersson are both extremely helpful in all aspects of acoustic analysis of ultrasonic signals, including the basic principles involved. They can be reached by e‐mail from these sites.
  This site describes several systems available for automated analysis of other behaviors in addition to the automated recording of USV.
  This site displays a wealth of hard‐to‐find products for physiological and microsurgical studies on rats and mice.
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