The nonapeptide arginine vasotocin (AVT) and its mammalian homologue arginine vasopressin are well known for their role in the modulation of several intraspecific social behaviours, such as social approach/withdrawal and aggression. Recently, we suggested that AVT might also be important in the regulation of interspecific social behaviours as it modulates interspecific cooperative behaviours in the Indo-Pacific bluestreak cleaner wrasse Labroides dimidiatus. AVT injections decreased cleaners' propensity to engage in cleaning interactions with their clients, suggesting that high levels of this peptide compromise the cleaners' cooperative motivation. Therefore, we hypothesise that low endogenous levels of AVT are a prerequisite for the expression of interspecific cleaning behaviour in cleaner wrasses, since it allows them to approach and interact with interspecific individuals, and that this should be reflected in their AVT neuronal phenotype. Here we test this hypothesis by comparing the AVT neuronal phenotypes of two phylogenetically closely related species that live in similar environments but diverge in the expression of interspecific cooperative behaviour: an obligate cleaner wrasse (L. dimidiatus) and a non-cleaner corallivore wrasse (Labrichthys unilineatus). The two species are predicted to differ in their AVT neuronal phenotypes as a reflection of their ability/inability to approach and interact with interspecific individuals, with cleaners presenting smaller and/or less numerous AVT-immunoreactive (ir) neurons. A sex difference in AVT neuronal phenotypes was also predicted because males of both species appear to be more aggressive than females. As described for most of the other teleost species, AVT-ir neurons were restricted to the preoptic area, and in agreement with our first prediction cleaners presented smaller and less numerous AVT-ir neurons in the gigantocellular preoptic area (gPOA) compared to non-cleaners. Contrary to our second prediction, AVT neuronal phenotypes did not differ between sexes in either species, but differences in other features of the AVT system cannot be ruled out. In summary, the results presented here suggest a putative role for AVT gPOA neurons in the ability of a cleaner wrasse to approach and interact with a client, through their projections to extrahypothalamic brain areas.

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