Chen Bingbing, Wang Shengnan, Inglis Briauna Marie, Ding Hao, Suo Angbaji, Qiu Shuai, Duan Yanchao, Li Xi, Li Shanshan, Sun Wendell Q, Si Wei
Institute of Biothermal Science and Technology, School of Medical Instruments and Food Engineering, University of Shanghai for Science and Technology, Shanghai, China.
State Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, China.
Front Physiol. 2021 Oct 4;12:719346. doi: 10.3389/fphys.2021.719346. eCollection 2021.
Antifreeze protein III (AFP III) is used for the cryopreservation of germ cells in various animal species. However, the exact mechanism of its cryoprotection is largely unknown at the molecular level. In this study, we investigated the motility, acrosomal integrity, and mitochondrial membrane potential (MMP), as well as proteomic change, of cynomolgus macaque sperm after cryopreservation. Sperm motility, acrosomal integrity, and MMP were lower after cryopreservation ( < 0.001), but significant differences in sperm motility and MMP were observed between the AFP-treated sperm sample (Cryo+AFP) and the non-treated sample (Cryo-AFP) ( < 0.01). A total of 141 and 32 differentially expressed proteins were, respectively, identified in cynomolgus macaque sperm cryopreserved without and with 0.1 μg/ml AFP III compared with fresh sperm. These proteins were mainly involved in the mitochondrial production of reactive oxygen species (ROS), glutathione (GSH) synthesis, and cell apoptosis. The addition of AFP III in the sperm freezing medium resulted in significant stabilization of cellular molecular functions and/or biological processes in sperm, as illustrated by the extent of proteomic changes after freezing and thawing. According to the proteomic change of differentially expressed proteins, we hypothesized a novel molecular mechanism for cryoprotection that AFP III may reduce the release of cytochrome c and thereby reduce sperm apoptosis by modulating the production of ROS in mitochondria. The molecular mechanism that AFP III acts with sperm proteins for cellular protection against cryoinjuries needs further study.
抗冻蛋白III（AFP III）用于多种动物物种生殖细胞的冷冻保存。然而，其冷冻保护的确切分子机制在很大程度上尚不清楚。在本研究中，我们调查了食蟹猴精子冷冻保存后的活力、顶体完整性、线粒体膜电位（MMP）以及蛋白质组变化。冷冻保存后精子活力、顶体完整性和MMP降低（<0.001），但在AFP处理的精子样本（Cryo+AFP）和未处理的样本（Cryo-AFP）之间观察到精子活力和MMP存在显著差异（<0.01）。与新鲜精子相比，在分别不含和含有0.1μg/ml AFP III的条件下冷冻保存的食蟹猴精子中，分别鉴定出141种和32种差异表达蛋白。这些蛋白主要参与线粒体活性氧（ROS）的产生、谷胱甘肽（GSH）合成和细胞凋亡。精子冷冻培养基中添加AFP III导致精子细胞分子功能和/或生物学过程显著稳定，这通过冻融后蛋白质组变化的程度得以体现。根据差异表达蛋白的蛋白质组变化，我们推测了一种新的冷冻保护分子机制，即AFP III可能通过调节线粒体中ROS的产生来减少细胞色素c的释放，从而减少精子凋亡。AFP III与精子蛋白共同作用以保护细胞免受冷冻损伤的分子机制需要进一步研究。
