Urinary Proteins of Female Domestic Dog (Canis familiaris) during Ovarian Cycle.

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In this study, we aim to evaluate the urinary proteins of female domestic dogs in the estrus and anestrus phases to evidence the presence of non-volatile chemical signals and to elucidate their identities. We collected urine samples from eight female dogs in the estrus and anestrus phases. A total of 240 proteins were identified in the urine samples using liquid chromatography-mass spectrometry (LC-MS analysis). The comparison of the proteins revealed a significant difference between the estrus and anestrus urine. We identified proteins belonging to the lipocalin family of canines (beta-lactoglobulin-1 and beta-lactoglobulin-2, P33685 and P33686, respectively), one of whose function was the transport of pheromones and which was present only in the estrus urine samples. Moreover, proteins such as Clusterin (CLU), Liver-expressed antimicrobial peptide 2 (LEAP2), and Proenkephalin (PENK) were more abundant in the estrus urine when compared to the anestrus urine. LEAP2 was recently described as a ghrelin receptor antagonist and implicated in regulating food intake and body weight in humans and mice. Proenkephalin, a polypeptide hormone cleaved into opioid peptides, was also recognized as a candidate to determine kidney function. As of yet, none of these have played a role in chemical communication. Clusterin, an extracellular chaperone protecting from protein aggregation implicated in stress-induced cell apoptosis, is a plausible candidate in chemical communication, which is a claim that needs to be ascertained further. Data are available via ProteomeXchange with the identifier PXD040418.", "claims": [{"id": "claim-method", "statement": "The statistical significance of quantification values for proteins was tested with background-based multiple t-testing.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:37104448", "locator": "sections[15] \"2.5. Statistical Methods\" paragraph 2", "quote": "The statistical significance of quantification values for proteins was tested with background-based multiple t-testing."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The data that support the findings of this study are available from the corresponding author (M.D.) upon reasonable request.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:37104448", "locator": "sections[25] \"notes\" paragraph 1", "quote": "The data that support the findings of this study are available from the corresponding author (M.D.) upon reasonable request."}], "state": "complete"}, {"id": "claim-target-result", "statement": "The comparison of the proteins revealed a significant difference between the estrus and anestrus urine.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:37104448", "locator": "sections[0] \"Abstract\" paragraph 2", "quote": "The comparison of the proteins revealed a significant difference between the estrus and anestrus urine."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "The comparison of the proteins revealed a significant difference between the estrus and anestrus urine.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:37104448"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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