大白公猪精浆外泌体miRNA表征鉴定及功能分析

Characterization,identification and functional analysis of miRNA in seminal plasma exosomes of Yorkshire boar

[目的]精浆外泌体(Seminal plasma exosome,spEX)在精子成熟、凋亡、受精中起着至关重要的作用。本文旨在探究种公猪spEX miRNA表达及miRNA在精子成熟及功能维持过程中的潜在调控作用。[方法]提取大白公猪spEX并进行电镜分析、粒径分析、标志性蛋白表达分析和miRNA高通量测序。[结果]成功分离出spEX,利用miRNA测序共鉴定出329个spEX miRNA。对高表达miRNA进行靶基因预测和功能富集分析,结果表明spEX miRNA在射精、P53信号通路、前列腺癌、细胞对DNA损伤刺激的反应、负调控凋亡过程、顶体膜结合、受精等通路中均发挥了潜在调控作用。[结论]本研究为spEX miRNA在调控精子活力和精子受精作用方面提供基础数据,并为精液保存调控机制的研究提供参考。

【Objective】Seminal plasma exosomes(spEXs)play a crucial role in sperm maturation,apoptosis and fertilization.This study was aimed to explore the miRNA expression of boar spEXs and the potential regulatory role of miRNA in sperm maturation and functional maintenance.【Method】The spEXs from Yorkshire boar semen were isolated and the exosomes were identified through transmission electron microscopy,nanoparticle tracking analysis,marker protein expression analysis and high-throughput miRNA sequencing.【Result】The spEXs were successfully isolated.A total of 329 spEX miRNAs were identified by miRNA sequencing.Through target gene prediction and functional enrichment analysis of highly expressed miRNAs,it was concluded that spEX miRNAs played potential regulatory roles in sperm ejaculation,P53 signaling pathway,prostate cancer,cell response to DNA damage stimuli,negative regulation of apoptosis,acrosome membrane binding,fertilization,etc.【Conclusion】This study provides basic data for spEX miRNAs regulating sperm motility and sperm fertilization,and provides references for studying the regulation mechanism of semen preservation.