Research report
Female urine-induced male mice ultrasonic vocalizations, but not scent-marking, is modulated by social experience

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Abstract

Despite the evidence for a communicative function of rodent scent marks and ultrasonic vocalizations, relatively little is known about the impact of social factors on these two forms of communication. Here, we tested the effects of two important social factors, prior exposure to a female and freshness of female urine, on male scent marks and ultrasonic vocalizations elicited by female urine. We also asked whether a recently reported strain difference between the highly social strain C57BL/6J (B6) and the mouse model of autism BTBR T+tf/J (BTBR) herein is specifically seen in response to female urine or also detectable in response to male urine traces. Results show that the emission of female urine-elicited ultrasonic vocalizations was dependent on previous female experience, while scent-marking behavior was not affected. A positive correlation was detected between scent-marking behavior and ultrasonic calling in the most biologically relevant context, male mice exposed to fresh female urine after female experience. Correlations were less prominent or missing in less biologically relevant contexts, e.g. in male mice exposed to fresh female urine without previous female experience, indicating that previous female experience is affecting both the emission of female urine-elicited ultrasonic vocalizations and the correlation between olfactory and acoustic communication. The strain difference in scent-marking behavior and ultrasonic calling between B6 and BTBR appears to be specific to female urine-elicited behavior as it was not seen in response to male urine traces, highlighting the relevance of the social context in which mouse communication is evaluated.

Introduction

Rodents communicate primarily via olfactory and acoustic signals. A major source of olfactory communicative signals is contained in scent marks. Scent-marking, the deposition of urine traces in strategic environmental locations, is used by mice to demarcate territories, orchestrate aggressive behavior, recognize individuals, maintain family organization, communicate danger, and to attract mates [1], [2], [3], [4]. Male mice scent marks serve to indicate territorial boundaries [5], [6], advertising the mouse's ability to dominate it and hence the likelihood to maintain the territory against intruders [7], [8], [9] and aid in the orchestration of aggressive behavior [10], [12], [13], [14], [15]. They also allow the recognition of individuals and hence maintenance of family organization [16], [17]. Male scent marks attract female mice [18], [19], [20], [21], [22], [23], [24]. Furthermore, they induce estrous [25] and accelerate the onset of puberty [26]. Scent marks from unfamiliar males can interrupt the establishment of pregnancy in female mice [27].

In addition to the deposition of scent marks, rodents emit ultrasonic vocalizations as acoustic communicative signals [28], [29], [30], [31], [32]. Dependent on species, age, sex and affective state, different types of ultrasonic vocalizations can be detected that appear to serve different communicative purposes. Pup isolation-induced ultrasonic vocalizations elicit maternal retrieval behavior in mice and rats [33], [34], [35], [36]. Anxiety-induced ultrasonic vocalizations serve an alarm function and elicit freezing behavior in rats [37], [38], [39], [40]. Interaction-induced ultrasonic vocalizations elicit social approach behavior in mice and rats [39], [41]. Males exposed to females or female urine emit ultrasonic vocalizations that facilitate mating behavior in mice and rats [42], [43], [44], [45], [46], [47], [48], [49], [50]. In mice, ultrasonic calling is specifically seen in males exposed to female urine as male urine is ineffective [51], [52], [53], [54], [55], [56], [57].

Despite the evidence for a communicative function of rodent scent marks and ultrasonic vocalizations, relatively little is known about the impact of social factors on scent-marking and ultrasonic vocalizations. One of the best known phenomena of social modulation of male scent-marking in response to female scents is its dependence on social status. Dominant males scent mark more than subordinate males [11], [19], [58], [59] and social defeat leads to a reduction in male scent-marking [60]. Importantly, females prefer scent marks deposited by dominant males [19], [21]. Relatively little, however, is known about the social status for the production of ultrasonic vocalizations in response to female scents [61], [62]. Here, one of the best known phenomena of social modulation is the dependence on previous female experience [51], [52], [53], [63], [64], [65], [66]. Typically, male mice do not emit ultrasonic vocalizations when exposed to female urine if they have not been in contact with females previously. However, some findings appear to be inconsistent with a critical familiarity or learning component. In some reports, male mice that had no previous female experience reliably emitted ultrasonic vocalizations to female urine, although mostly to a lower extent [46], [57], [64], [65]. These inconsistencies in male ultrasonic calling might be due to a second modulating factor, freshness of the female urine [64], [65], [66]. For both factors, however, little is known about their effect on the time course of the male's ultrasonic vocalization response.

The first aim of the present study was a more detailed characterization of the effects of these two factors, prior exposure to a female and freshness of the female urine, on male ultrasonic vocalizations elicited by female urine. The second aim was to test whether those two factors affect olfactory communication as well as acoustic responses. The third aim was to test the hypothesis that vocalization and scent-marking responses to female urine are linked. We recently reported that mice who deposit high numbers of scent marks in response to female urine emit more ultrasonic vocalizations than mice who deposit low numbers of scent marks [67]. This link between the two systems was seen under optimal conditions, namely male mice with previous female experience exposed to fresh female urine. The present experiments were designed to understand whether this link holds true under less optimal conditions.

The fourth aim of the present study addressed social cues mediating scent-marking between two inbred strains of mice which differ on sociability. We reported that scent-marking as well as the emission of ultrasonic vocalizations by male mice in response to female urine is dependent on the sociability of the strain [67]. C57BL/6J (B6) is a highly social strain of mice, while BTBR T+tf/J (BTBR) is a low sociability strain that may represent a mouse model of autism [68], [69], [70], [71], [72], [73], [74]. We found that adult male BTBR deposited fewer scent marks and emitted extremely low numbers of ultrasonic vocalizations as compared to adult male B6. The question addressed in the present study is whether this strain difference is specific to exposure to female urine, relevant to sexual mating [18], [19], [20], [21], [22], [23], [24]. Male/male scent countermarking represents a different domain of mouse communication, relevant to territoriality [5], [7], [8], [9], [10], [11], [12], [13], [14], [15] and dominance hierarchies [12], [58], [59], [60]. We therefore tested whether a similar strain difference is seen in scent countermarking in response to male urine.

Section snippets

Animals and housing

Subjects were adult male C57BL/6J (B6) and BTBR T+tf/J (BTBR) mice. For the male subject + female urine spot experiment, N = 45 male B6 mice were used. For the male subject + male urine scent marks experiment, N = 40 male B6 and N = 40 male BTBR mice were used. Breeding pairs were purchased from The Jackson Laboratory (Bar Harbor, ME, USA) and bred at the National Institute of Mental Health in Bethesda, MD, USA. About 2 weeks after pairing for breeding, the females were individually housed and

Male subject + female urine spot: ultrasonic vocalizations

As shown in Fig. 2, emission of ultrasonic vocalizations was dependent on experimental condition in male B6 mice. When comparing the total number of ultrasonic vocalizations emitted during the entire 5 min of female B6 urine exposure, a significant effect of experimental condition was detected by One-Way ANOVA (F2,44 = 3.336, p = 0.045; Fig. 2A). Males with previous female experience exposed to fresh female urine emitted 438.33 ± 109.39 calls, while much lower call numbers were detected in the two

Discussion

Emission of female urine-elicited ultrasonic vocalizations in adult male mice was dependent on previous female experience. Male mice with a previous 5 min exposure to an adult female mouse approximately 1 week before the test session displayed an overall higher ultrasonic vocalization response than male mice with no previous exposure to an adult female mouse. Further, the latency to vocalize was shorter and the rate of vocalizations was more sustained in males with previous exposure to a female.

Acknowledgment

Supported by the National Institute of Mental Health Intramural Research Program.

References (82)

  • J.W. Scott et al.

    Behavioral and electrophysiological responses of female mice to male urine odors

    Physiol Behav

    (1970)
  • T.J. Rich et al.

    The competing countermarks hypothesis: reliable assessment of competitive ability by potential mates

    Anim Behav

    (1999)
  • M. Wöhr et al.

    Activation of limbic system structures by replay of ultrasonic vocalization in rats, Chapter 4.2

  • M. Wöhr et al.

    Effect of altricial pup ultrasonic vocalization on maternal behavior, Chapter 5.2

  • S.M. Brudzynski et al.

    Behavioural responses of laboratory rats to playback of 22 kHz ultrasonic calls

    Physiol Behav

    (1995)
  • G.D. Sales

    The effect of 22 kHz calls and artificial 38 kHz signals on activity in rats

    Behav Proc

    (1991)
  • M. Wöhr et al.

    Ultrasonic calling during fear conditioning in the rat: no evidence for an audience effect

    Anim Behav

    (2008)
  • K. Musolf et al.

    Ultrasonic courtship vocalizations in wild house mice, Mus musculus musculus

    Anim Behav

    (2010)
  • S.M. Pomerantz et al.

    Female behavior is affected by male ultrasonic vocalizations in house mice

    Physiol Behav

    (1983)
  • D.A. Thomas et al.

    Male-produced postejaculatory vocalizations and the mating behaviour of estrous female rats

    Behav Neural Biol

    (1982)
  • G. Dizinno et al.

    Ultrasonic vocalizations by male mice (Mus musculus) to female sex pheromone: experimental determinants

    Behav Biol

    (1978)
  • J. Nyby et al.

    Pheromonal regulation of male mouse ultrasonic courtship (Mus musculus)

    Anim Behav

    (1977)
  • J. Nyby et al.

    Elicitation of male mouse ultrasounds: bladder urine and aged urine from females

    Physiol Behav

    (1980)
  • J. Nyby et al.

    Male mouse (Mus musculus) ultrasonic vocalizations to female urine: why is heterosexual experience necessary?

    Behav Neural Biol

    (1983)
  • L.C. Drickamer

    Urine marking and social dominance in male house mice (Mus musculus domesticus)

    Behav Proc

    (2001)
  • L.A. Lumley et al.

    Social stress effects on territorial marking and ultrasonic vocalizations in mice

    Physiol Behav

    (1999)
  • F.R. D’Amato

    Courtship ultrasonic vocalizations and social status in mice

    Anim Behav

    (1991)
  • J. Nyby et al.

    Social status and ultrasonic vocalizations of male mice

    Behav Biol

    (1976)
  • J.C. Maggio et al.

    Experience-based vocalization of male mice to female chemosignals

    Physiol Behav

    (1983)
  • M.L. Sipos et al.

    An ephemeral sex pheromone in the urine of female house mice (Mus domesticus)

    Behav Neural Biol

    (1992)
  • M.L. Sipos et al.

    An ephemeral pheromone of female house mice: perception via the main and accessory olfactory systems

    Physiol Behav

    (1995)
  • M.L. Sipos et al.

    An ephemeral sex pheromone in the urine of female house mice (Mus domesticus): pheromone fade-out time

    Physiol Behav

    (1993)
  • V.J. Bolivar et al.

    Assessing autism-like behaviour in mice: variations in social interactions among inbred strains

    Behav Brain Res

    (2007)
  • S.S. Moy et al.

    Mouse behavioral tasks relevant to autism: phenotypes of 10 inbred strains

    Behav Brain Res

    (2007)
  • M. Yang et al.

    Social approach behaviors are similar on conventional versus reverse lighting cycles, and in replications across cohorts, in BTBR T+tf/J, C57BL/6J, and vasopressin receptor 1B mutant mice

    Front Behav Neurosci

    (2007)
  • M. Yang et al.

    Social deficits in BTBR T+tf/J mice are unchanged by cross-fostering with C57BL/6J mothers

    Int J Dev Neurosci

    (2007)
  • H. Arakawa et al.

    Scent marking behavior in male C57BL/6J mice: sexual and developmental determination

    Behav Brain Res

    (2007)
  • J. Nyby et al.

    Postpubertal experience establishes signal value of mammalian sex odor

    Behav Biol

    (1978)
  • J. Nyby et al.

    Intracranial androgenic and estrogenic stimulation of male-typical behaviors in house mice (Mus domesticus)

    Horm Behav

    (1992)
  • H. Arakawa et al.

    Social features of scent-donor mice modulate scent marking of C57BL/6J recipient males

    Behav Brain Res

    (2009)
  • J.A. Maruniak et al.

    Urinary marking in male house mice: responses to novel environmental and social stimuli

    Physiol Behav

    (1974)
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