Faculty of Science, School of Life and Environmental Sciences, The University of Sydney, NSW 2006, Australia.
Faculty of Science, School of Life and Environmental Sciences, The University of Sydney, NSW 2006, Australia.
J Proteomics. 2018 Jun 15;181:73-82. doi: 10.1016/j.jprot.2018.04.001. Epub 2018 Apr 5.
Cryopreservation causes significant lethal and sub-lethal damage to spermatozoa. In order to improve freezing outcomes, a comprehensive understanding of sub-lethal damage is required. Cryopreservation induced changes to sperm proteins have been investigated in several species, but few have employed currently available state of the art, data independent acquisition mass spectrometry (MS) methods. We used the SWATH LC-MS method to quantitatively profile proteomic changes to ram spermatozoa following exposure to egg yolk and cryopreservation. Egg yolk contributed 15 proteins to spermatozoa, including vitellogenins, apolipoproteins and complement component C3. Cryopreservation significantly altered the abundance of 51 proteins. Overall, 27 proteins increased (e.g. SERPINB1, FER) and 24 proteins decreased (e.g. CCT subunits, CSNK1G2, TOM1L1) in frozen thawed ram spermatozoa, compared to fresh spermatozoa. Chaperones constituted 20% of the proteins lost from spermatozoa following cryopreservation. These alterations may interfere with both normal cellular functioning and the ability of frozen thawed spermatozoa to appropriately respond to stress. This is the first study to apply SWATH mass spectrometry techniques to characterise proteins contributed by egg yolk based freezing media and to profile cryopreservation induced proteomic changes to ram spermatozoa.
This study profiles changes to the sperm proteome induced by exposure to egg yolk based media and the process of cryopreservation, and the biological consequences are discussed.
冷冻保存会对精子造成显著的致死和亚致死损伤。为了改善冷冻效果，需要全面了解亚致死损伤。已经在几种物种中研究了冷冻保存诱导的精子蛋白变化，但很少有研究采用目前可用的最先进的、无依赖于标签的数据获取质谱（MS）方法。我们使用 SWATH LC-MS 方法定量分析了暴露于卵黄和冷冻保存后公羊精子的蛋白质组变化。卵黄为精子贡献了 15 种蛋白质，包括卵黄蛋白、载脂蛋白和补体成分 C3。冷冻保存显著改变了 51 种蛋白质的丰度。总体而言，与新鲜精子相比，冷冻解冻的公羊精子中 27 种蛋白质增加（如 SERPINB1、FER），24 种蛋白质减少（如 CCT 亚基、CSNK1G2、TOM1L1）。冷冻保存后，从精子中丢失的伴侣蛋白构成了 20%。这些变化可能会干扰精子的正常细胞功能和冷冻解冻精子对适当应激的反应能力。这是首次应用 SWATH 质谱技术来描述基于卵黄的冷冻介质贡献的蛋白质，并对公羊精子冷冻保存诱导的蛋白质组变化进行分析的研究。
本研究分析了暴露于基于卵黄的介质和冷冻保存过程中精子蛋白质组的变化，并讨论了其生物学后果。
