National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, Guangdong, China; Lingnan Guangdong Laboratory of Modern Agriculture, Guangzhou, 510642, Guangdong, China; College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, 471023, Henan, China.
National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, Guangdong, China; Lingnan Guangdong Laboratory of Modern Agriculture, Guangzhou, 510642, Guangdong, China.
Theriogenology. 2024 Jan 1;213:79-89. doi: 10.1016/j.theriogenology.2023.09.026. Epub 2023 Oct 2.
Extracellular vesicles (EVs) are abundant in body fluid and are critical in cell interaction. Seminal plasma contains numerous EVs which affecting sperm function via transferring regulatory cargoes to the sperm. However, the mechanism of seminal plasma extracellular vesicles (SP-EVs) is still not clear. The present study aimed to isolate the boar SP-EVs and explore its potential function, then identify the key protein involved in SP-EVs and sperms interaction, and elucidate mechanism of SP-EVs protein on sperms. Here, we successfully isolated and concentrated boar SP-EVs, the SP-EVs showed a typical vesicle structure under transmission electron microscopy, most of their diameters range between 50 and 200 nm and express EVs biomarkers CD9 and CD63. We proved that SP-EVs could inhibit sperm acrosome reaction and in vitro fertility. Through a data-independent acquisition analysis of protein profiles of noncapacitated sperms, normal capacitated sperms and SP-EVs treated capacitated sperms, we identified that EZRIN was one of the active proteins that participated in SP-EVs and sperms interaction. Furthermore, we tested that the inhibition of EZRIN could promote boar sperm fertility, which is in consistence with the function of SP-EVs. The results may facilitate future research of SP-EVs on sperm function and male infertility.
细胞外囊泡（EVs）在体液中丰富存在，在细胞间相互作用中起着关键作用。精浆中含有大量的 EVs，通过将调节性 cargo 转移到精子上，影响精子功能。然而，精浆细胞外囊泡（SP-EVs）的机制仍不清楚。本研究旨在分离猪的 SP-EVs 并探索其潜在功能，然后鉴定参与 SP-EVs 和精子相互作用的关键蛋白，并阐明 SP-EVs 蛋白对精子的作用机制。在这里，我们成功地分离并浓缩了猪的 SP-EVs，SP-EVs 在透射电子显微镜下显示出典型的囊泡结构，其直径大多在 50 至 200nm 之间，并表达 EVs 标志物 CD9 和 CD63。我们证明 SP-EVs 可以抑制精子顶体反应和体外生育能力。通过对非获能精子、正常获能精子和 SP-EVs 处理的获能精子的蛋白质图谱进行非依赖性采集分析，我们鉴定出 EZRIN 是参与 SP-EVs 和精子相互作用的活性蛋白之一。此外，我们测试了抑制 EZRIN 可以促进猪精子的生育能力，这与 SP-EVs 的功能一致。这些结果可能有助于未来对 SP-EVs 对精子功能和男性不育的研究。
