Dalmazzo Andressa, de Souza Ramos Angrimani Daniel, Losano João Diego A, Rocha Carolina C, Sobrinho Carlos A B, Chinait Gurgel João Rafael, Monteiro Pacheco Pedro Ivo, Minazaki Claudia Kiyomi, Crusco Silvia E, Nichi Marcilio, Barnabe Valquíria H
University of São Paulo,College of Veterinary Medicine and Animal Science,Department of Animal Reproduction, Cidade Universitária, São Paulo,Brazil.
Zygote. 2019 Feb;27(1):17-24. doi: 10.1017/S0967199418000576. Epub 2018 Dec 5.
SummaryThe aim of this study was to compare different concentrations of soy lecithin (LEC0.01%, LEC0.05% and LEC0.1%) with egg yolk (Control) in cooling extenders during the storage of semen at 5ºC for 5 days. Twelve dogs (n = 12) were selected, and semen was cooled and assessed after 2, 24, 48, 72, 96 or 120 h. At each time point, sperm were analyzed for kinetic patterns (using computer-assisted sperm analysis), mitochondrial activity (3'3- diaminobenzidine assay), lipid peroxidation (TBARS assay), DNA fragmentation (SCSA®) and plasma and acrosome membrane integrity (eosin/nigrosin and fast green/rose Bengal stains, respectively). The Control group (1814.4 ± 197.2) presented the highest rates of lipid peroxidation at 120 h. Conversely, progressive motility (42.8 ± 4%), linearity (45.4 ± 1%), and VAP (88 ± 3%) were higher in the Control group. In addition, there was lower mitochondrial activity in the Control group at 72 h. Therefore, our data show that lecithin used at these concentrations was not able to maintain sperm viability at as high qualities as would egg yolk. Moreover, the decrease in high mitochondrial activity and the persistence of sperm motility may indicate a compensatory mechanism in canine spermatozoa (i.e., glycolytic pathway). Furthermore, these higher lipid peroxidation indexes could indicate the necessity for future therapy using extenders and antioxidants over a long cooling time for dog sperm.
摘要
本研究的目的是比较在5℃下储存精液5天期间,不同浓度的大豆卵磷脂(LEC0.01%、LEC0.05%和LEC0.1%)与蛋黄(对照组)在冷却稀释液中的效果。选取了12只犬(n = 12),在2、24、48、72、96或120小时后对精液进行冷却和评估。在每个时间点,对精子进行动力学模式分析(使用计算机辅助精子分析)、线粒体活性检测(3'3-二氨基联苯胺检测)、脂质过氧化检测(硫代巴比妥酸反应物检测)、DNA片段化检测(精子染色质结构分析)以及血浆和顶体膜完整性检测(分别采用伊红/黑色素和固绿/孟加拉玫瑰红染色)。对照组(1814.4 ± 197.2)在120小时时脂质过氧化率最高。相反,对照组的前向运动率(42.8 ± 4%)、直线性(45.4 ± 1%)和平均路径速度(88 ± 3%)较高。此外,对照组在72小时时线粒体活性较低。因此,我们的数据表明,这些浓度的卵磷脂不能像蛋黄那样维持精子的高质量活力。而且,高线粒体活性的降低和精子活力的持续存在可能表明犬精子存在一种补偿机制(即糖酵解途径)。此外,这些较高的脂质过氧化指标可能表明未来需要使用稀释液和抗氧化剂对犬精子进行长时间冷却治疗。