背景背景来自输卵管液(oEV)的细胞外囊泡(EV)在各种生殖过程中发挥关键作用,包括精子获能,施肥,早期胚胎发育。目的从发情周期的不同阶段(晚期卵泡,LF;黄体早期,EL;黄体中期,ML;黄体晚期,LL)并研究它们对精子功能的影响。方法分离poEV,特点,并标记以评估它们与公猪精子的结合。poEV对精子运动的影响,生存能力,顶体状态,蛋白激酶A磷酸化(PKAs),酪氨酸磷酸化(Tyr-P),并对体外生育率进行了分析。关键结果poEV被观察为圆形或杯状膜包围的囊泡。统计分析表明,POEV在大小上没有显著差异,数量,或发情周期各阶段的蛋白质浓度。然而,LFpoEV表现出更高的与精子结合的亲和力。用EL治疗,ML,和LLpoEV导致精子进行性运动性和总运动性降低。此外,在存在LF的情况下,PKA水平降低,EL,和MLPoEV,而Tyr-P水平在组间没有差异。LFpoEV还降低了精子穿透率和每个穿透卵母细胞的精子数量(P结论来自发情周期不同阶段的poEV通过与精子相互作用在精子功能中起调节作用,影响运动和获能,并参与精子-卵母细胞的相互作用。提示LF和LLpoEV的不同作用表明,poEV可能用作IVF系统中的添加剂来调节精子-卵母细胞的相互作用。
Context Extracellular vesicles (EVs) derived from the oviductal fluid (oEVs) play a critical role in various reproductive processes, including sperm capacitation, fertilisation, and early embryo development. Aims To characterise porcine oEVs (poEVs) from different stages of the estrous cycle (late follicular, LF; early luteal, EL; mid luteal, ML; late luteal, LL) and investigate their impact on sperm functionality. Methods poEVs were isolated, characterised, and labelled to assess their binding to boar spermatozoa. The effects of poEVs on sperm motility, viability, acrosomal status, protein kinase A phosphorylation (pPKAs), tyrosine phosphorylation (Tyr-P), and in in vitro fertility were analysed. Key results poEVs were observed as round or cup-shaped membrane-surrounded vesicles. Statistical analysis showed that poEVs did not significantly differ in size, quantity, or protein concentration among phases of the estrous cycle. However, LF poEVs demonstrated a higher affinity for binding to sperm. Treatment with EL, ML, and LL poEVs resulted in a decrease in sperm progressive motility and total motility. Moreover, pPKA levels were reduced in presence of LF, EL, and ML poEVs, while Tyr-P levels did not differ between groups. LF poEVs also reduced sperm penetration rate and the number of spermatozoa per penetrated oocyte (P Conclusions poEVs from different stages of the estrous cycle play a modulatory role in sperm functionality by interacting with spermatozoa, affecting motility and capacitation, and participating in sperm-oocyte interaction. Implications The differential effects of LF and LL poEVs suggest the potential use of poEVs as additives in IVF systems to regulate sperm-oocyte interaction.