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

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.

SUBMITTER: Barabas AJ

PROVIDER: S-EPMC6879570 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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

Barabas Amanda J AJ Aryal Uma K UK Gaskill Brianna N BN

Scientific reports 20191126 1

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 materi ...[more]

PMID: 31772257

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Project description:Excessive home cage aggression often results in severe injury and subsequent premature euthanasia of male laboratory mice. Aggression can be reduced by transferring used nesting material during cage cleaning, which is thought to contain aggression appeasing odors from the plantar sweat glands. However, neither the composition of plantar sweat nor the deposits on used nesting material have been evaluated. The aims of this study were to (1) identify and quantify volatile compounds deposited in the nest site and (2) determine if nest and sweat compounds correlate with social behavior. Home cage aggression and affiliative behavior were evaluated in 3 strains: SJL, C57BL/6N, and A/J. Individual social rank was assessed via the tube test, because ranking may influence compound levels. Sweat and urine from the dominant and subordinate mouse in each cage, plus cage level nest samples were analyzed for volatile compound content using gas chromatography-mass spectrometry. Behavior data and odors from the nest, sweat, and urine were statistically analyzed with separate principal component analyses (PCA). Significant components, from each sample analysis, and strain were run in mixed models to test if odors were associated with behavior. Aggressive and affiliative behaviors were primarily impacted by strain. However, compound PCs were also impacted by strain, showing that strain accounts for any relationship between odors and behavior. C57BL/6N cages displayed the most allo-grooming behavior and had high scores on sweat PC1. SJL cages displayed the most aggression, with high scores on urine PC2 and low scores on nest PC1. These data show that certain compounds in nesting material, urine, and sweat display strain specific patterns which match strain specific behavior patterns. These results provide preliminary information about the connection between home cage compounds and behavior. Salient compounds will be candidates for future controlled studies to determine their direct effect on mouse social behavior.

Project description:Home cage aggression causes poor welfare in male laboratory mice and reduces data quality. One of the few proven strategies to reduce aggression involves preserving used nesting material at cage change. Volatile organic compounds from the nesting material and several body fluids not only correlate with less home cage aggression, but with more affiliative allo-grooming behavior. To date, these compounds have not been tested for a direct influence on male mouse social behavior. This study aimed to determine if 4 previously identified volatile compounds impact home cage interactions. A factorial design was used with cages equally split between C57BL/6N and SJL male mice (N = 40). Treatments were randomly assigned across cages and administered by spraying one compound solution on each cage's nesting material. Treatments were refreshed after day 3 and during cage change on day 7. Home cage social behavior was observed throughout the study week and immediately after cage change. Several hours after cage change, feces were collected from individual mice to measure corticosterone metabolites as an index of social stress. Wound severity was also assessed after euthanasia. Measures were analyzed with mixed models. Compound treatments did not impact most study measures. For behavior, SJL mice performed more aggression and submission, and C57BL/6N mice performed more allo-grooming. Wound severity was highest in the posterior region of both strains, and the middle back region of C57BL/6N mice. Posterior wounding also increased with more observed aggression. Corticosterone metabolites were higher in C57BL/6N mice and in mice treated with 3,4-dimethyl-1,2-cyclopentanedione with more wounding. These data confirm previous strain patterns in social behavior and further validates wound assessment as a measure of escalated aggression. The lack of observed treatment effects could be due to limitations in the compound administration procedure and/or the previous correlation study, which is further discussed.

Dataset Information

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

Publications

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

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