title: Protein modification in the post-mating spermatophore of the signal crayfish Pacifastacus leniusculus: insight into the tyrosine phosphorylation in a non-motile spermatozoon.
abstract: After mating, spermatophores of signal crayfish are stored on the body of the female for a period before fertilization. This study compared the post-mating protein profile and pattern of protein tyrosine phosphorylation of the signal crayfish spermatophore to that of the freshly ejaculated spermatophore and found substantial differences. Two major bands of tyrosine-phosphorylated proteins of molecular weights 10 and 50kDa were observed in the freshly ejaculated spermatophore of the signal crayfish. While the tyrosine-phosphorylated protein band with molecular weight 10kDa was formed by protein(s) of similar pH, the band with molecular weight of 50kDa consisted of proteins of varying pH. In the post-mating spermatophore, the band with molecular weight of 50kDa was not detected, and an increase in the level of protein tyrosine phosphorylation was observed in the 10kDa band. The microtubular radial arms of the spermatozoon showed a positive reaction to an anti-tyrosine antibody conjugated with gold particles in both the freshly ejaculated and post-mating spermatophores. In conclusion, the male gamete of the signal crayfish undergoes molecular modification during post-mating storage on the body of the female including changes in the level of protein expression and protein tyrosine phosphorylation. Structural similarity of the radial arms in the crayfish immotile spermatozoon with flagellum, which is the main site of protein tyrosine phosphorylation in the mammalian motile spermatozoa, raises questions regarding evolution and function of such organelles across the animal kingdom that must be addressed in the future studies.
authors.affiliation: University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zátiší 728/II, 38925 Vodňany, Czech Republic. Electronic address: niksirat@frov.jcu.cz.
authors.forename: Hamid
authors.initials: H
authors.lastname: Niksirat
authors.affiliation: Institute of Parasitology, Biology Centre of the ASCR and Faculty of Science, University of South Bohemia in České Budějovice, Branišovská 31, České Budějovice 37005, Czech Republic.
authors.forename: Marie
authors.initials: M
authors.lastname: Vancová
authors.affiliation: Department of Immunotechnology, Hus 406, Medicon Village, Lund University, 22381, Lund, Sweden.
authors.forename: Liselotte
authors.initials: L
authors.lastname: Andersson
authors.affiliation: Department of Immunotechnology, Hus 406, Medicon Village, Lund University, 22381, Lund, Sweden.
authors.forename: Peter
authors.initials: P
authors.lastname: James
authors.affiliation: University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zátiší 728/II, 38925 Vodňany, Czech Republic.
authors.forename: Antonín
authors.initials: A
authors.lastname: Kouba
authors.affiliation: University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zátiší 728/II, 38925 Vodňany, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Kozák
mesh_terms: D000818:Animals; D003400:Astacoidea / Q000502:physiology*; D003307:Copulation; D005260:Female; D008297:Male; D010766:Phosphorylation / Q000502:physiology; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D013081:Sperm Motility / Q000502:physiology*; D013093:Spermatogonia / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D011506:Proteins; D014443:Tyrosine
keywords: Microtubular radial arm; Spermatozoon capacitation; Tyrosine-phosphorylation; Ultrastructural localization