SERPINA5 Protein in Cumulus-Oocyte Complexes Increases the Fertilisation Ability of Mouse Sperm
SERPINA5 Protein in Cumulus-Oocyte Complexes Increases the Fertilisation Ability of Mouse Sperm
复制标题
卵丘-卵母细胞复合物中的 SERPINA5 蛋白可提高小鼠精子的受精能力
DOI：
10.1007/s43032-022-00867-5
复制
发表时间：
2022-02-22
影响因子：
2.9
通讯作者：
Yao，Bing
中科院分区：
文献类型：
--
作者：
Cao，Siyuan;Qian，Zhang;Yao，Bing
获得高质量的精子是提高辅助生殖技术（ART）成功率的关键。尽管卵丘-卵母细胞复合体（COCs）分泌的细胞因子与精子表面受体结合以改善精子质量，但将小鼠COCs添加到人输卵管液（HTF）培养基中对精子获能的影响尚未被探索。将8周龄的ICR小鼠COC加入HTF培养基中并粉碎以获得后修饰的HTF培养基。与HTF培养液相比，改良后的HTF培养液体外获能后受精率和与透明质酸结合的精子数显著增加（P< 0.01）。蛋白质组学和Western blotting分析表明，经改良的HTF培养液体外获能后，精子中SERPINA 5的水平显著升高（P< 0.05）。免疫组化染色结果显示SERPINA 5蛋白在小鼠卵丘细胞中有表达。体外获能后，在改良HTF培养液中加入SERPINA 5抗体，阻断SERPINA 5的作用，使受精率和与透明质酸结合的精子数显著降低（P< 0.05）。在HTF培养液中加入重组小鼠SERPINA 5蛋白（1 ~ 2 μg/ml），受精率和精子结合率均显著提高（P< 0.01）。此外，在人精液冷冻前加入重组人SERPINA 5蛋白（5 μg/ml）。与不添加SERPINA 5蛋白相比，添加SERPINA 5蛋白后精子形态正常率和顶体完整率显著提高（P< 0.05）。本研究为体外获能过程中优化精子质量提供了参考方法。
Obtaining high-quality sperm is key to improving the success rate of assisted reproductive technology (ART). Although cytokines secreted by cumulus-oocyte complexes (COCs) bind to sperm surface receptors to improve sperm quality, the effects of adding mouse COCs to human tubal fluid (HTF) medium on sperm capacitation have not yet been explored. Eight-week-old ICR mouse COCs were added to HTF medium and crushed to obtain the post-modified HTF medium. Compared with using HTF medium, the fertilisation rate and number of sperm combined with the zona pellucida significantly increased after in vitro capacitation using the post-modified HTF medium (P< 0.01). Proteomic and Western blotting analyses showed that the level of SERPINA5 in sperm increased significantly following in vitro capacitation with the post-modified HTF medium (P< 0.05). Immunohistochemical staining analysis demonstrated that SERPINA5 protein was expressed in mouse cumulus cells. A SERPINA5 antibody was added in the post-modified HTF medium to block the effects of SERPINA5 after in vitro capacitation, which significantly decreased the fertilisation rate and the number of sperm combined with the zona pellucida (P< 0.05). Recombinant mouse SERPINA5 protein (1 ~ 2 μg/ml) was added to HTF medium and the fertilisation rate and the number of sperm combined with the zona pellucida significantly increased (P< 0.01). Moreover, recombinant human SERPINA5 protein (5 μg/ml) was added before human semen freezing. Compared with adding no SERPINA5 protein, the percentage of normal sperm morphology and the intact acrosome significantly increased (P< 0.05). Our study provides a reference method for optimising sperm quality in the process of in vitro capacitation.