Fish sperm cryopreservation is a well-established technique allowing for artificial insemination on a commercial scale. The extent of proteome alterations in seminal plasma and sperm due to cryopreservation, however, is not known. This study was conducted to evaluate the effect of cryopreservation on motility variables of sterlet Acipenser ruthenus sperm and to detect the differences in protein profiles of fresh and cryopreserved sterlet sperm and seminal plasma. Fresh sperm had 89 ± 3% motility and 160 ± 14 μm/s curvilinear velocity at 15 s post-activation. The motility rate of cryopreserved sperm (37 ± 5%) was less at 15 s post-activation. No difference (ANOVA; P > 0.05) in mean curvilinear velocity of fresh and cryopreserved sperm was detected. The protein profiles of seminal plasma and sperm were characterized using comparative proteomics to determine the influence of cryopreservation. Six altered protein spots in seminal plasma and thirteen altered spots in sperm were detected in fresh and thawed sperm. Subsequent protein characterization suggested that the proteins identified were involved in sperm metabolism, cytoskeleton, and stress response. The results broaden the understanding of the effects of cryopreservation and identify the proteins associated with cryo-injury. These data may help to determine the function of altered proteins and provide new insights into improving sperm cryopreservation.

CAS Key Laboratory of Biobased Materials Qingdao Institute of Bioenergy and Bioprocess Technology Chinese Academy of Sciences No 189 Songling Road Qingdao 266101 China

University of South Bohemia in Ceske Budejovice Faculty of Fisheries and Protection of Waters South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses Research Institute of Fish Culture and Hydrobiology Zatisi 728 2 38925 Vodnany Czech Republic

University of South Bohemia in Ceske Budejovice Faculty of Fisheries and Protection of Waters South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses Research Institute of Fish Culture and Hydrobiology Zatisi 728 2 38925 Vodnany Czech Republic; Sino Czech Joint Laboratory for Fish Conservation and Biotechnology Yangtze River Fisheries Research Institute Chinese Academy of Fishery Sciences Wuhan China

University of South Bohemia in Ceske Budejovice Faculty of Science Institute of Chemistry and Biochemistry Branisovska 1760 37005 Ceske Budejovice Czech Republic

University of South Bohemia in Ceske Budejovice Faculty of Science Institute of Chemistry and Biochemistry Branisovska 1760 37005 Ceske Budejovice Czech Republic; Biology Centre of Academy of Sciences of the Czech Republic Institute of Parasitology Branisovska 31 37005 Ceske Budejovice Czech Republic

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Protective role of antifreeze proteins on sterlet (Acipenser ruthenus) sperm during cryopreservation