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Identification of protein pheromones that promote aggressive behaviour

Abstract

Mice use pheromones, compounds emitted and detected by members of the same species, as cues to regulate social behaviours such as pup suckling, aggression and mating1. Neurons that detect pheromones are thought to reside in at least two separate organs within the nasal cavity: the vomeronasal organ (VNO) and the main olfactory epithelium (MOE)2. Each pheromone ligand is thought to activate a dedicated subset of these sensory neurons. However, the nature of the pheromone cues and the identity of the responding neurons that regulate specific social behaviours are largely unknown. Here we show, by direct activation of sensory neurons and analysis of behaviour, that at least two chemically distinct ligands are sufficient to promote male–male aggression and stimulate VNO neurons. We have purified and analysed one of these classes of ligand and found its specific aggression-promoting activity to be dependent on the presence of the protein component of the major urinary protein (MUP) complex, which is known to comprise specialized lipocalin proteins bound to small organic molecules1,3,4. Using calcium imaging of dissociated vomeronasal neurons (VNs), we have determined that the MUP protein activates a sensory neuron subfamily characterized by the expression of the G-protein Gαo subunit (also known as Gnao) and Vmn2r putative pheromone receptors (V2Rs). Genomic analysis indicates species-specific co-expansions of MUPs and V2Rs, as would be expected among pheromone-signalling components. Finally, we show that the aggressive behaviour induced by the MUPs occurs exclusively through VNO neuronal circuits. Our results substantiate the idea of MUP proteins as pheromone ligands that mediate male–male aggression through the accessory olfactory neural pathway.

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Figure 1: Male urine contains two aggression pheromones.
Figure 2: HMW aggression activity is dependent on MUPs.
Figure 3: MUPs activate a subset of VNs that express Gαo.
Figure 4: MUP activation is specific to VNs.

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Acknowledgements

We thank F. Papes, U. Mueller, A. Patapoutian, K. Baldwin and C. Zucker for discussions and critical reading of the manuscript. This work was supported by funding from the NIDCD, Pew Charitable Trust, Skaggs Institute, and Helen Dorris Foundation (to L.S.), a National Institute on Deafness and Other Communication Disorders (NIDCD) pre-doctoral fellowship (T.F.M.) and The Basque Government Post-Doctoral Research Fellowship (P.C.).

Author Contributions The behavioural analysis was performed by P.C., J.R.C. and L.S.; calcium imaging was done by P.C. and T.F.M.; and biochemical purification of MUPs by T.F.M. and advised by B.F.C. SBT synthesis was performed by A.S. and B.F.C.; construct preparation and immunostaining by P.C., T.F.M. and K.F.; and genomic analysis by D.W.L. All authors participated in data analysis and writing of the manuscript.

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Correspondence to Lisa Stowers.

Supplementary information

Supplementary Information

This file includes Supplementary Figures 1 -5 with Legends, Supplementary Methods and additional references. The Supplementary Figures show wide range different control experiments for the calcium imaging, dose response to urine in behavior (SFig1), MUP purification by FPLC (SFig2), validation of SBT displacement by GC/MS (FigS3), ISH for Gi and Go antibodies (FigS4), and a neighbor-joining tree of MUP-like proteins from some terrestrial vertebrates (FigS5). (PDF 575 kb)

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Chamero, P., Marton, T., Logan, D. et al. Identification of protein pheromones that promote aggressive behaviour. Nature 450, 899–902 (2007). https://doi.org/10.1038/nature05997

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