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, 389 25 Vodňany, Czech Republic; Department of Parasitology, Faculty of Science, Charles University, BIOCEV, Průmyslová 595, 252 50 Vestec, Czech Republic.
BioCeV - Institute of Microbiology of the CAS, Průmyslová 595, 252 50 Vestec, Czech Republic; University of South Bohemia in Ceske Budejovice, Faculty of Science, Branisovska 1760, 37005 Ceske Budejovice, Czech Republic; Orekhovich Institute of Biomedical Chemistry, Pogodinskaja str. 10, 119191, Moscow, Russia.
Anim Reprod Sci. 2023 Apr;251:107213. doi: 10.1016/j.anireprosci.2023.107213. Epub 2023 Mar 14.
Using cryopreservation techniques can increase the effectiveness of reproducing cultured fish species by ensuring a dependable supply of sperm, although the quality of the sperm could be impacted by the procedures involved. The goal of this study was to investigate the effect of purified seminal plasma transferrin (Tf), bovine serum albumin (BSA), and antifreeze protein (AFP) types I and III at 1 µg mL on relevant characteristics of cryopreserved sperm from common carp Cyprinus carpio. We compared oxidative stress indices, antioxidant activity, and DNA fragmentation of fresh sperm to that frozen with extender only or with Tf, BSA, or AFP types I and III. Fresh sperm had significantly lower levels of thiobarbituric acid reactive substances (TBARS) compared to samples that underwent cryopreservation without protein treatment, which resulted in 0.54 ± 0.06 nmol/10 cells of TBARS. Carbonyl derivatives of proteins (CP) decreased significantly (ANOVA; P > 0.05) in carp sperm with addition of Tf, AFPI, and AFPIII. Significant differences in superoxide dismutase (SOD), glutathione reductase (GR), and glutathione peroxidase (GPx) activity were seen in sperm supplemented with Tf, BSA, AFPI, and AFPIII from those without. Significantly less DNA damage, expressed as percent tail DNA (11.56 ± 1.34) and olive tail moment (0.59 ± 0.13), was recorded in samples cryopreserved with Tf. The findings indicated that addition of Tf, BSA, AFPI, or AFPIII to cryopreservation medium is beneficial to sperm preservation. The mechanisms through which these proteins act positively on sperm need to be further investigated.
使用冷冻保存技术可以通过确保可靠的精子供应来提高养殖鱼类物种的繁殖效果,尽管所涉及的程序可能会影响精子的质量。本研究的目的是研究在 1 µg/mL 时,纯化的精液转铁蛋白 (Tf)、牛血清白蛋白 (BSA) 和抗冻蛋白 (AFP) 类型 I 和 III 对鲤鱼精子冷冻保存相关特性的影响。我们比较了新鲜精子和仅用冷冻液、Tf、BSA 或 AFP 类型 I 和 III 冷冻精子的氧化应激指标、抗氧化活性和 DNA 碎片化。与未用蛋白质处理冷冻的样品相比,新鲜精子的硫代巴比妥酸反应物质 (TBARS) 水平明显较低,为 0.54±0.06 nmol/10 个细胞。在添加 Tf、AFPI 和 AFPIII 后,鲤鱼精子中的蛋白质羰基衍生物 (CP) 显著减少 (ANOVA;P>0.05)。在添加 Tf、BSA、AFPI 和 AFPIII 的精子中,超氧化物歧化酶 (SOD)、谷胱甘肽还原酶 (GR) 和谷胱甘肽过氧化物酶 (GPx) 的活性有显著差异。用 Tf 冷冻保存的精子的 DNA 损伤明显较少,表现为尾部 DNA 百分比 (11.56±1.34) 和橄榄尾部矩 (0.59±0.13)。添加 Tf、BSA、AFPI 或 AFPIII 到冷冻保存液中有利于精子保存。这些蛋白质对精子产生积极作用的机制需要进一步研究。