Protein Expression of Cortisol Metabolising Enzymes in the Boar Reproductive Tract

P151 Protein Expression of Cortisol Metabolising Enzymes in the Boar Reproductive Tract V Sharp, AE Michael and LM Thurston Royal Veterinary College, University College London, London, UK The stress hormone cortisol is implicated in a wide range of reproductive processes including semen quality. Cortisol is interconverted with cortisone by isoforms of the enzyme 11beta hydroxysteroid dehydrogenase (11bHSD). Investigations in our laboratory have demonstrated that the boar reproductive tract secretes modulators of 11bHSD activity. We hypothesise that modulation of 11bHSD in the boar tract alters the environment in which the sperm mature, thus modifying sperm quality and the ability to withstand cold temperature storage. The aims of this study were to establish whether 11bHSD1 and 11bHSD2 proteins are expressed in the boar reproductive tract. Boar testis, epididymis, vas deferens, bulbourethral glands and penis were obtained from slaughterhouse material from 7 animals. Tissues were homogenised in a Tris-glycerol-SDS lysis buffer containing a protease inhibitor cocktail. Proteins were resolved by SDS-PAGE and 11bHSD protein expression was assessed using Western blotting. The immunopurified 11bHSD1 antibody recognised a protein band of 32 kDa in porcine liver (positive control) and in boar testis, epididymis and vas deferens, but not in porcine kidney (negative control), bulbourethral glands or penis. Alternatively, the immunopurified 11bHSD2 antibody recognised a protein band of 57 kDa in all reproductive tissues assessed. In conclusion, 11bHSD protein was expressed at several sites in the boar reproductive tract. Cortisol generation by 11bHSD is known to influence both sodium concentration and pH, thus modulation of this protein may result in changes to the luminal environment ultimately affecting sperm quality.