Li Shanshan, Ao Lei, Yan Yaping, Jiang Jiang, Chen Bingbing, Duan Yanchao, Shen Fei, Chen Jinbao, Inglis Briauna, Ni Renmin, Ji Weizhi, Si Wei
1Yunnan Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan China.
Kunming Sino-UK Angel Women's & Children's Hospital, Kunming, Yunnan China.
Clin Proteomics. 2019 Jun 19;16:24. doi: 10.1186/s12014-019-9244-2. eCollection 2019.
Although sperm cryopreservation has been widely used in human reproductive medicine as an integral infertility management in infertility clinics and for banking sperm in sperm banks, the freezing/thawing protocols are not optimal. The freezing and thawing processes result in changes at both structural and molecular levels, some even detrimental, in human sperm when compared with fresh sperm. The change of sperm proteins after cryopreservation may play negative roles for fertilization and early embryo development. Conventionally, cryostraws (CS) and cryovials (CV) are the most widely used cryopreservation carriers (CPCs) for human sperm cryopreservation accompanied with the use of egg yolk free commercial media. However, the influence of cryopreservation on the proteomic profile of human sperm preserved with the two CPCs is unknown. Therefore the purpose of the present study was to compare the frozen-thawed motility, investigate the proteomic profile of human sperm cryopreserved with the two types of CPCs, and identify the susceptible proteins that play key roles for sperm function and fertility.
The present study compared the cryosurvival of human sperm frozen with the two different CPCs and identified the sperm proteomic changes by using the isobaric tags for relative and absolute quantification labeling technique coupled with 2D LC-MS/MS analysis after freezing and thawing.
Our results indicated that sperm cryopreserved with CV showed higher values for percentage of motile sperm and forward activity rate than those with CS. Compared to fresh sperm, 434 and 432 proteins were differentially identified in human sperm cryopreserved with CS and CV, respectively.
The proteomic profiles of human sperm are greatly affected by cryopreservation with either type of CPC. GO analysis revealed that most of the differentially identified sperm proteins enriched in the extracellular membrane-bounded organelles, cytoplasm and cytosol. In addition, 106 susceptible proteins having known identities related to sperm functions were identified. In general, cryovial seems to be the preferred CPC for human sperm cryopreservation based on the post-thaw motility parameters and the effect on sperm proteomic profiles. These results are beneficial for the insight into the understanding of the cryoinjury mechanism of sperm and the development of human sperm cryopreservation strategies.
尽管精子冷冻保存已在人类生殖医学中广泛应用，作为不孕不育诊所不孕症综合管理的一部分以及精子库中精子储存的方法，但冷冻/解冻方案并非最优。与新鲜精子相比，冷冻和解冻过程会导致人类精子在结构和分子水平上发生变化，有些甚至是有害的。冷冻保存后精子蛋白质的变化可能对受精和早期胚胎发育产生负面影响。传统上，冷冻细管（CS）和冻存管（CV）是人类精子冷冻保存中使用最广泛的冷冻保存载体（CPC），同时使用不含蛋黄的商业培养基。然而，冷冻保存对用这两种CPC保存的人类精子蛋白质组图谱的影响尚不清楚。因此，本研究的目的是比较冷冻解冻后的活力，研究用两种类型的CPC冷冻保存的人类精子的蛋白质组图谱，并确定对精子功能和生育能力起关键作用的易感蛋白。
本研究比较了用两种不同CPC冷冻的人类精子的冷冻存活率，并在冷冻和解冻后使用相对和绝对定量标记技术结合二维液相色谱-串联质谱分析来确定精子蛋白质组的变化。
我们的结果表明，用CV冷冻保存的精子的活动精子百分比和向前活动率值高于用CS冷冻保存的精子。与新鲜精子相比，用CS和CV冷冻保存的人类精子分别差异鉴定出434和432种蛋白质。
用任何一种CPC进行冷冻保存都会极大地影响人类精子的蛋白质组图谱。基因本体分析显示，大多数差异鉴定的精子蛋白富集在细胞外膜结合细胞器、细胞质和细胞溶质中。此外，还鉴定出106种与精子功能相关的已知身份的易感蛋白。总体而言，基于解冻后活力参数和对精子蛋白质组图谱的影响，冻存管似乎是人类精子冷冻保存的首选CPC。这些结果有助于深入了解精子冷冻损伤机制和人类精子冷冻保存策略的发展。
