engleski

USE OF HIGH-RESOLUTION QUANTITATIVE PROTEOMIC APPROACH TO IDENTIFY CHANGES IN SALIVARY PROTEINS AFTER AN INDUCED EJACULATION IN DOGS

Introduction: The conjunction of modern proteomic diagnostic techniques, such as Tandem Mass Tags (TMT), with saliva samples allow to achieve comparative quantitative analysis of this fluid and the identification of new metabolic pathways and possible biomarkers. Individual analytes, such as alpha-amylase or cortisol, has been studied in saliva after an ejaculation in dogs. However, no complete studies about changes in salivary proteome have been reported. Objectives: Investigate the possible changes in salivary proteome profile of dogs during ejaculation. Methods: Saliva specimens were collected from eight healthy male beagles 30 min before (preejaculation) and immediately after the ejaculation (post-ejaculation) provoked by digital manipulation, using a latex semen collection cone (artificial vagina). Saliva samples were subjected to reduction, alkylation, digestion and labelled using 6-plex TMT reagents before further LC–MS/MS analysis. The results were then processed by the Gene Ontology (GO) analysis. Results: The TMT analysis allowed for the identification of new ejaculation-related metabolic pathways and revealed 33 of recognized peptides showed significant differences (p<0.05) after ejaculation. Of those proteins, 17 were down-regulated while 16 were up-regulated. The most affected metabolic pathways significantly involved identified by GO analysis were antimicrobial humoral response and nucleosome assembly (Figure 1). Conclusions: TMT-based proteomic approach allowed identification of new salivary proteins that change in concentration after ejaculation in dogs. However, further studies should be made to clarify the mechanisms of these changes and if they could be used as biomarkers.

Saliva, Dog, Ejaculation, Proteomics

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