Lucio C F, Brito M M, Angrimani Dsr, Belaz Kra, Morais D, Zampieri D, Losano Jda, Assumpção Meoa, Nichi M, Eberlin M N, Vannucchi C I
Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
Thomson Mass Spectrometry Laboratory, University of Campinas, Campinas, São Paulo, Brazil.
Reprod Domest Anim. 2017 Apr;52 Suppl 2:208-213. doi: 10.1111/rda.12860. Epub 2016 Nov 3.
The fatty acid composition of the sperm membrane is an important factor involved in the overall sperm quality, including motility. However, in the canine species, the exact composition of the plasma membrane is still unknown. Therefore, the purpose of this study was to evaluate the plasma membrane lipid composition of motile sperm cells and to compare it with asthenospermic samples, as an attempt to determine possible involvements of membrane lipids in dog sperm cell motility. The sperm-rich fraction of ten mature dogs was collected, and samples were subjected to density gradient centrifugation by Percoll , in order to separate motile and asthenospermic samples. Processed semen samples were evaluated for sperm motility, plasma and acrosome membrane integrity, mitochondrial activity and susceptibility to oxidative stress. Lipid plasma membrane composition was identified by mass spectrometry (MALDI-MS). The motile sperm samples presented the following phospholipids in a high frequency in the plasma membrane: phosphatidylcholine 38:4 (composed of stearic and arachidonic fatty acids), phosphatidylcholine 36:1 (stearic and oleic fatty acids), phosphatidylethanolamine 34:4 (myristic and arachidonic fatty acids), glycerophosphatidic acid 36:4 (palmitic and arachidonic fatty acids), phosphatidylcholine 40:4 plasmanyl and phosphatidylcholine 40:5 plasmenyl. Furthermore, no lipid markers were found in the asthenospermic samples. Results also indicate that differences on plasma membrane composition between motile and asthenospermic samples are crucial factors for determining sperm motility, sperm functionality and susceptibility to oxidative stress. In conclusion, plasma membrane lipid composition varies considerable between motile and asthenospermic samples. Therefore, lipid markers of sperm motility can be considered, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylcholine plasmanyl, phosphatidylcholine plasmenyl and phosphatidic acid.
精子膜的脂肪酸组成是影响精子整体质量(包括活力)的一个重要因素。然而,在犬类中,质膜的确切组成仍然未知。因此,本研究的目的是评估活动精子细胞质膜的脂质组成,并将其与弱精子症样本进行比较,以确定膜脂在犬精子细胞活力中可能的作用。收集了十只成年犬的富含精子部分,并通过Percoll进行密度梯度离心,以分离活动精子和弱精子症样本。对处理后的精液样本进行精子活力、质膜和顶体膜完整性、线粒体活性以及对氧化应激敏感性的评估。通过质谱法(基质辅助激光解吸电离质谱)鉴定质膜脂质组成。活动精子样本在质膜中呈现出以下高频率的磷脂:磷脂酰胆碱38:4(由硬脂酸和花生四烯酸组成)、磷脂酰胆碱36:1(硬脂酸和油酸)、磷脂酰乙醇胺34:4(肉豆蔻酸和花生四烯酸)、甘油磷脂酸36:4(棕榈酸和花生四烯酸)、磷脂酰胆碱40:4质体酰和磷脂酰胆碱40:5质膜酰。此外,在弱精子症样本中未发现脂质标志物。结果还表明,活动精子和弱精子症样本之间质膜组成的差异是决定精子活力、精子功能和对氧化应激敏感性的关键因素。总之,活动精子和弱精子症样本之间质膜脂质组成差异很大。因此,可以考虑将精子活力的脂质标志物,如磷脂酰胆碱、磷脂酰乙醇胺、质体酰磷脂酰胆碱、质膜酰磷脂酰胆碱和磷脂酸。