Abstract

Chemosignals mediate both intra- and inter-specific communication in most mammals. Pheromones elicit stereotyped reactions in conspecifics, whereas kairomones provoke a reaction in an allospecific animal. For instance, predator kairomones elicit anticipated defensive responses in preys. The aim of this work was to test the behavioral responses of female mice to two chemosignals: 2-heptanone (2-HP), a putative alarm pheromone, and 2,4,5-trimethylthiazoline (TMT), a fox-derived putative kairomone, widely used to investigate fear and anxiety in rodents. The banana-like odorant isoamyl acetate (IA), unlikely to act as a chemosignal, served as a control odorant. We first presented increasing amounts of these odorants in consecutive days, in a test box in which mice could explore or avoid them. Female mice avoided the highest amounts of all three compounds, with TMT and IA eliciting avoidance at lower amounts (3.8 pmol and 0.35 μmol, respectively) than 2-HP (35 μmol). All three compounds induced minimal effects in global locomotion and immobility in this set up. Further, mice detected 3.5 pmol of TMT and IA in a habituation–dishabituation test, so avoidance of IA started well beyond the detection threshold. Finally, both TMT and IA, but not 2-HP, induced conditioned place avoidance and increased immobility in the neutral compartment during a contextual memory test. These data suggest that intense odors can induce contextual learning irrespective of their putative biological significance. Our results support that synthetic predator-related compounds (like TMT) or other intense odorants are useful to investigate the neurobiological basis of emotional behaviors in rodents. Since intense odorants unlikely to act as chemosignals can elicit similar behavioral reactions than chemosignals, we stress the importance of using behavioral measures in combination with other physiological (e.g., hormonal levels) or neural measures (e.g., immediate early gene expression) to establish the ethological significance of odorants.

Highlights

  • Rodents are widely used in studies on the neurobiological basis of emotional behaviors

  • Odorants We used three odorants: 2,5-dihydro-2,4,5-trimethylthiazoline (TMT; Contech, Victoria, Canada), a putative kairomone found in fox feces and detected by the main olfactory system and the Grueneberg ganglion (Brechbühl et al, 2013), 2-heptanone (2HP, Sigma-Aldrich, Schnelldorf, Germany), a putative mouse pheromone detected by the main and accessory olfactory epithelia (Thompson et al, 2004; Xu et al, 2005), and isoamyl acetate (IA, Panreac Quimica SA, Barcelona, Spain), a control odorant detected by the main olfactory system (Xu et al, 2005) and unlikely to participate in intra or interspecific communication (Root et al, 2014)

  • None of the behavioral measures varied across days in these mice exposed to PBS alone, so any behavioral changes in the mice exposed to the different odorants could be attributed to the effects of the stimuli. (Supplementary Material, Experiment S2)

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Summary

Introduction

Rodents are widely used in studies on the neurobiological basis of emotional behaviors. Alarm pheromones are substances released by an injured or threatened animal and detected by conspecifics (Gutiérrez-García et al, 2007; Brechbühl et al, 2013), whereas predator kairomones elicit defensive responses in preys (for a review, see Fortes-Marco et al, 2013). The volatile chemical 2-heptanone (2-HP) has been proposed to act as an alarm substance in bees (Collins et al, 1989) and rats (Gutiérrez-García et al, 2007). This ketone is a component of the urine of rats, and its concentration is higher in stressed individuals. Alarm pheromones and kairomones might act using the same neural circuits conveying signals of danger (Brechbühl et al, 2013)

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