title: Characterization of proteolytic and anti-proteolytic activity involvement in sterlet spermatozoon maturation.
abstract: In sturgeon, the acquisition of the potential for motility activation called spermatozoon maturation takes place outside testes. This process can be accomplished in vitro by pre-incubation of immature testicular spermatozoa in seminal fluid collected from fully mature Wolffian duct sperm. Addition of trypsin inhibitor to the pre-incubation medium disrupts spermatozoon maturation. There are no available data for the role of proteolysis regulators in fish spermatozoon maturation, while their role is recognized in mammalian sperm maturation. The present study evaluated the involvement of seminal fluid proteases and anti-proteolytic activity in the sterlet spermatozoon maturation process. Casein and gelatin zymography and quantification of amidase and anti-proteolytic activity were conducted in sturgeon seminal fluid from Wolffian duct sperm and seminal fluid from testicular sperm, along with spermatozoon extracts from Wolffian duct spermatozoa, testicular spermatozoa, and testicular spermatozoa after in vitro maturation. We did not find significant differences in proteolytic profiles of seminal fluids from Wolffian duct sperm and ones from testicular sperm. Zymography revealed differences in spermatozoon extracts: Wolffian duct spermatozoon extracts were characterized by the presence of a broad proteolytic band ranging from 48 to 41 kDa, while testicular spermatozoon extracts did not show such activity until after in vitro maturation. The differences in amidase activity coincided with these results. It may not be the levels of proteolytic and anti-proteolytic activity per se, but the alterations in their interactions triggering a cascade of signaling events, that is crucial to the maturation process.
authors.affiliation: South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Zatisi 728/II, 389 25, Vodnany, Czech Republic. vdzyuba@frov.jcu.cz.
authors.forename: Viktoriya
authors.initials: V
authors.lastname: Dzyuba
authors.affiliation: Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Tuwima 10, 10-748, Olsztyn, Poland.
authors.forename: Mariola
authors.initials: M
authors.lastname: Słowińska
authors.affiliation: South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Zatisi 728/II, 389 25, Vodnany, Czech Republic.
authors.forename: Jacky
authors.initials: J
authors.lastname: Cosson
authors.affiliation: Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Tuwima 10, 10-748, Olsztyn, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Ciereszko
authors.affiliation: South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Zatisi 728/II, 389 25, Vodnany, Czech Republic.
authors.forename: Sergii
authors.initials: S
authors.lastname: Boryshpolets
authors.affiliation: Institute of Chemistry and Biochemistry, Faculty of Science, University of South Bohemia in Ceske Budejovice, Branisovska 1760, 37005, Ceske Budejovice, Czech Republic.|Institute of Parasitology, Biology Centre of ASCR, Branisovska 31, 37005, Ceske Budejovice, Czech Republic.
authors.forename: Ján
authors.initials: J
authors.lastname: Štĕrba
authors.affiliation: South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Zatisi 728/II, 389 25, Vodnany, Czech Republic.
authors.forename: Marek
authors.initials: M
authors.lastname: Rodina
authors.affiliation: South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Zatisi 728/II, 389 25, Vodnany, Czech Republic.
authors.forename: Otomar
authors.initials: O
authors.lastname: Linhart
authors.affiliation: South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Zatisi 728/II, 389 25, Vodnany, Czech Republic.
authors.forename: Borys
authors.initials: B
authors.lastname: Dzyuba
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D005399:Fishes / Q000502:physiology*; D008297:Male; D059748:Proteolysis; D013079:Sperm Maturation; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology*; D013737:Testis / Q000166:cytology; D014928:Wolffian Ducts / Q000166:cytology
chemical_list: D000581:Amidohydrolases; C019665:amidase
keywords: Amidase activity; Anti-proteolytic activity; Casein and gelatin zymography; Siberian sturgeon sperm; Spermatozoon maturation; Sterlet sperm