Zhang Yuxin, Liu Dongqing, Wang Qinghua, Ruan Qingxin, Hua Sijie, Zhang Weiwei, Yang Sen, Meng Zining
State Key Laboratory of Biocontrol, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory of Aquatic Economic Animals, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
College of Food Science and Technology, Guangdong Ocean University (Yangjiang Campus), Yangjiang 529599, China.
Animals (Basel). 2024 Sep 12;14(18):2652. doi: 10.3390/ani14182652.
Spermatozoa cryopreservation protocols have been established for yellow catfish (), but cryopreservation can still cause cellular damage and affect spermatozoa viability and fertility. Therefore, the aim of this paper was to evaluate the effects of adding or not adding cryoprotectants during low-temperature storage on the ultrastructural damage, oxidative damage, and DNA damage of thawed yellow catfish spermatozoa. The mixed semen of three male yellow catfish was divided into a fresh spermatozoa group, a frozen spermatozoa group (DMSO) with a cryoprotectant (10% DMSO), and a frozen spermatozoa group without a cryoprotectant (DMSO). Ultrastructural of the spermatozoa after thawing were observed under an electron microscope and the spermatozoa were assayed for SOD, MDA, and T-AOC enzyme activities, as well as for DNA integrity. In terms of movement parameters, compared with DMSO, the addition of DMSO has significantly improved sperm motility, curve line velocity (VCL), and straight line velocity (VSL). The ultrastructural results showed that although thawed spermatozoa exhibited increased damage than fresh spermatozoa, 10% DMSO effectively reduced the damage to the plasma membrane, mitochondria, and flagellum of spermatozoa by cryopreservation, and most of the spermatozoa were preserved with intact structure. The results of oxidative damage showed that compared with frozen spermatozoa, 10% DMSO significantly increased the activities of SOD and T-AOC enzymes and clearly reduced the activity of the MDA enzyme. The antioxidant capacity of spermatozoa was improved, lipid peroxidation was reduced, and the oxidative damage caused by cryopreservation was mitigated. The DNA integrity of spermatozoa showed that 10% DMSO clearly reduced the DNA fragmentation rate. In conclusion, 10% DMSO can effectively reduce the ultrastructural damage, oxidative damage, and DNA damage of yellow catfish spermatozoa during cryopreservation; it can also further optimize the cryopreservation protocol for yellow catfish spermatozoa. Meanwhile, it also provides a theoretical basis for the future optimization of the cryopreservation protocols.
黄颡鱼精子冷冻保存方案已经确立，但冷冻保存仍会导致细胞损伤，影响精子活力和受精能力。因此，本文旨在评估低温保存过程中添加或不添加冷冻保护剂对解冻后的黄颡鱼精子超微结构损伤、氧化损伤及DNA损伤的影响。将3尾雄性黄颡鱼的混合精液分为新鲜精子组、添加冷冻保护剂（10%二甲基亚砜，DMSO）的冷冻精子组和不添加冷冻保护剂的冷冻精子组。解冻后的精子在电子显微镜下观察超微结构，并检测精子的超氧化物歧化酶（SOD）、丙二醛（MDA）和总抗氧化能力（T-AOC）酶活性以及DNA完整性。在运动参数方面，与不添加DMSO相比，添加DMSO显著提高了精子活力、曲线运动速度（VCL）和直线运动速度（VSL）。超微结构结果显示，虽然解冻后的精子比新鲜精子损伤增加，但10% DMSO有效减少了冷冻保存对精子质膜、线粒体和鞭毛的损伤，大部分精子结构完整。氧化损伤结果表明，与冷冻精子相比，10% DMSO显著提高了SOD和T-AOC酶活性，明显降低了MDA酶活性。精子的抗氧化能力提高，脂质过氧化减少，冷冻保存引起的氧化损伤减轻。精子的DNA完整性表明，10% DMSO明显降低了DNA片段化率。综上所述，10% DMSO能有效减少黄颡鱼精子冷冻保存过程中的超微结构损伤、氧化损伤和DNA损伤；还能进一步优化黄颡鱼精子冷冻保存方案。同时，也为今后优化冷冻保存方案提供了理论依据。
