Xin Miaomiao, Shaliutina-Kolesova Anna, Sterba Jan, Konik Peter, Boryshpolets Sergii, Rodina Marek, Li Ping, Nian Rui, Linhart Otomar
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/II, 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 Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zatisi 728/II, 38925 Vodnany, Czech Republic.
Anim Reprod Sci. 2018 May;192:280-289. doi: 10.1016/j.anireprosci.2018.03.025. Epub 2018 Mar 27.
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.
鱼类精子冷冻保存是一项成熟的技术，可实现商业规模的人工授精。然而，冷冻保存对精浆和精子蛋白质组的改变程度尚不清楚。本研究旨在评估冷冻保存对俄罗斯鲟精子活力参数的影响，并检测新鲜和冷冻保存的俄罗斯鲟精子及精浆蛋白质谱的差异。新鲜精子在激活后15秒时的活力为89±3%，曲线速度为160±14μm/s。冷冻保存的精子在激活后15秒时的活力率较低（37±5%）。未检测到新鲜和冷冻保存精子的平均曲线速度有差异（方差分析；P>0.05）。使用比较蛋白质组学对精浆和精子的蛋白质谱进行表征，以确定冷冻保存的影响。在新鲜和解冻的精子中，检测到精浆中有6个蛋白斑点改变，精子中有13个斑点改变。随后的蛋白质表征表明，鉴定出的蛋白质参与精子代谢、细胞骨架和应激反应。这些结果拓宽了对冷冻保存影响的理解，并确定了与冷冻损伤相关的蛋白质。这些数据可能有助于确定改变蛋白质的功能，并为改善精子冷冻保存提供新的见解。
