Gelatinolytic and anti-trypsin activities in seminal plasma of common carp: relationship to blood, skin mucus and spermatozoa

Abstract

Proteases and protease inhibitors were detected in the seminal plasma, blood plasma, skin mucus and spermatozoa. Their molecular weights were estimated using SDS-PAGE under non-reducing conditions. The results demonstrate that the two main bands of anti-proteinase activity (APA) detected earlier in common carp seminal plasma with molecular weight of approximately 47 and 58 kDa are also present in other fluids. The intensity of staining was highest in blood and seminal plasma. The intensity in skin mucus was visible, but in a sperm extract it was faint. An additional fast migrating band (30 kDa) was observed only in seminal plasma. Highest APA was found in blood and seminal plasma followed by skin mucus. The activity in sperm extracts was low. The two serine-like proteases with molecular weight of 79 and 189 kDa in seminal and blood plasma have also been found. In skin mucus, protease of 79 kDa was also present. Metalloproteinases with molecular weights of 61 and 69 kDa were found in seminal and blood plasma but metalloproteinases of 44 and 38 kDa were observed only in seminal plasma. Although metalloproteinases, of molecular weight ranging between 61 and 75 kDa, were also visible in a sperm extract, the experimental approach used in this study did not allow unequivocal identification of unique proteases of spermatozoa. Blood plasma contains a serine protease and protease inhibitor not present in seminal plasma. Skin mucus also showed the profile of three unique proteases (two EDTA stimulated and one metalloproteinase). These results indicate that the analysis of proteases and their inhibitors makes it possible to distinguish contamination of milt with either blood or skin mucus. The physiological role of the detected protease-inhibitory system in fish seminal plasma is still unknown. It is possible that the protease inhibitor and proteases, unique for seminal plasma, are involved in a specific function of milt or testis (e.g. the control of spermatogenesis).