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      Proteome characterization of used nesting material and potential protein sources from group housed male mice, Mus musculus

      research-article
      1 , , 2 , 1
      Scientific Reports
      Nature Publishing Group UK
      Animal behaviour, Proteomics

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          Abstract

          Laboratory mice ( Mus musculus) communicate a variety of social messages through olfactory cues and it is often speculated that these cues are preserved in nesting material. Based on these speculations, a growing number of husbandry recommendations support preserving used nests at cage cleaning to maintain familiar odors in the new cage. However, the content of used nesting material has never been chemically analyzed. Here we present the first comprehensive proteome profile of used nesting material. Nests from cages of group housed male mice contain a variety of proteins that primarily originate from saliva, plantar sweat, and urine sources. Most notably, a large proportion of proteins found in used nesting material belong to major urinary protein (“MUP”) and odorant binding protein (“OBP”) families. Both protein families send messages about individual identity and bind volatile compounds that further contribute to identity cues. Overall, this data supports current recommendations to preserve used nesting material at cage cleaning to maintain odor familiarity.

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          Loss of sex discrimination and male-male aggression in mice deficient for TRP2.

          The mouse vomeronasal organ (VNO) is thought to mediate social behaviors and neuroendocrine changes elicited by pheromonal cues. The molecular mechanisms underlying the sensory response to pheromones and the behavioral repertoire induced through the VNO are not fully characterized. Using the tools of mouse genetics and multielectrode recording, we demonstrate that the sensory activation of VNO neurons requires TRP2, a putative ion channel of the transient receptor potential family that is expressed exclusively in these neurons. Moreover, we show that male mice deficient in TRP2 expression fail to display male-male aggression, and they initiate sexual and courtship behaviors toward both males and females. Our study suggests that, in the mouse, sensory activation of the VNO is essential for sex discrimination of conspecifics and thus ensures gender-specific behavior.
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            Regulation of salivary gland function by autonomic nerves.

            Oral homeostasis is dependent upon saliva and its content of proteins. Reflex salivary flow occurs at a low 'resting' rate and for short periods of the day more intense taste or chewing stimuli evoke up to ten fold increases in salivation. The secretion of salivary fluid and proteins is controlled by autonomic nerves. All salivary glands are supplied by cholinergic parasympathetic nerves which release acetylcholine that binds to M3 and (to a lesser extent) M1 muscarinic receptors, evoking the secretion of saliva by acinar cells in the endpieces of the salivary gland ductal tree. Most salivary glands also receive a variable innervation from sympathetic nerves which released noradrenaline from which tends to evoke greater release of stored proteins, mostly from acinar cells but also ductal cells. There is some 'cross-talk' between the calcium and cyclic AMP intracellular pathways coupling autonomic stimulation to secretion and salivary protein secretion is augmented during combined stimulation. Other non-adrenergic, non-cholinergic neuropeptides released from autonomic nerves evoke salivary gland secretion and parasympathetically derived vasointestinal peptide, acting through endothelial cell derived nitric oxide, plays a role in the reflex vasodilatation that accompanies secretion. Neuronal type, calcium-activated, soluble nitric oxide within salivary cells appears to play a role in mediating salivary protein secretion in response to autonomimetics. Fluid secretion by salivary glands involves aquaporin 5 and the extent to which the expression of aquaporin 5 on apical acinar cell membranes is upregulated by cholinomimetics remains uncertain. Extended periods of autonomic denervation, liquid diet feeding (reduced reflex stimulation) or duct ligation cause salivary gland atrophy. The latter two are reversible, demonstrating that glands can regenerate provided that the autonomic innervation remains intact. The mechanisms by which nerves integrate with salivary cells during regeneration or during salivary gland development remain to be elucidated.
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              Identification of protein pheromones that promote aggressive behaviour.

              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 mating. 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). 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 molecules. 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 Galpha(o) 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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                Author and article information

                Contributors
                abarabas@purdue.edu
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                26 November 2019
                26 November 2019
                2019
                : 9
                : 17524
                Affiliations
                [1 ]ISNI 0000 0004 1937 2197, GRID grid.169077.e, Department of Animal Science, , Purdue University, ; West Lafayette, IN 47907 USA
                [2 ]ISNI 0000 0004 1937 2197, GRID grid.169077.e, Purdue Proteomics Facility, , Purdue University, ; West Lafayette, IN 47907 USA
                Author information
                http://orcid.org/0000-0003-4543-1536
                Article
                53903
                10.1038/s41598-019-53903-x
                6879570
                31772257
                54489bd8-4c2f-4ef2-b1ef-441794302b84
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 18 June 2019
                : 4 November 2019
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                © The Author(s) 2019

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                animal behaviour,proteomics
                Uncategorized
                animal behaviour, proteomics

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