Small-extracellular vesicles and their microRNA cargo from porcine follicular fluids:the potential association with oocyte quality

作     者：Ahmed Gad Matej Murin Alexandra Bartkova Veronika Kinterova Katerina Marcollova Jozef Laurincik Radek Prochazka Ahmed Gad;Matej Murin;Alexandra Bartkova;Veronika Kinterova;Katerina Marcollova;Jozef Laurincik;Radek Prochazka

作者机构：Institute of Animal Physiology and GeneticsCzech Academy of Sciences27721 LiběchovCzech Republic Department of Animal ProductionFaculty of AgricultureCairo UniversityGiza 12613Egypt Department of Botany and GeneticsFaculty of Natural SciencesConstantine the Philosopher University in Nitra94901 NitraSlovakia. 

出 版 物：《Journal of Animal Science and Biotechnology》 (畜牧与生物技术杂志（英文版）)

年 卷 期：2022年第13卷第6期

页      面：1727-1742页

核心收录：

学科分类：0905[农学-畜牧学] 09[农学] 

基　　金：supported by the Institute of Animal Physiology and Genetics“IAPG-Matoušek Award 2020”Grant number:DRMA-2020-0002 the Ministry of Education,Youth and Sports of the Czech Republic,Operational Program Research,Development and Education,the project“EXCELLENCE in molecular aspects of the early development of vertebrates”Grant number:CZ.02.1.01/0.0/0.0/15_003/0000460. 

主　　题：Extracellular vesicles Follicular fuids MiRNA Oocyte quality Porcine 

摘      要：Background: Ovarian follicular fluids(FFs) contain several kinds of regulatory factors that maintain a suitable micro-environment for oocyte development. Extracellular vesicles(EVs) are among the factors that play essential roles in regulating follicle and oocyte development through their cargo molecules that include microRNAs(miRNAs). This study aimed to investigate small-EV(s-EV) miRNAs in porcine FFs and their potential association with oocyte quality.Methods: Individual aspirated oocytes were stained with lissamine green B stain(LB), a vital stain for oocyte quality, and each oocyte was classified as high-quality(unstained;HQ) or low-quality(stained;LQ). FFs corresponding to oocytes were pooled together into HQ and LQ groups. Small-EVs were isolated from FFs, characterized, and their miRNA cargo was identified using the Illumina Nova Seq sequencing platform. Additionally, s-EVs from the HQ and LQ groups were utilized to investigate their effect on oocyte development after co-incubation during in vitro maturation.Results: A total of 19 miRNAs(including miR-125b, miR-193a-5p, and miR-320) were significantly upregulated, while 23(including miR-9, miR-206, and miR-6516) were downregulated in the HQ compared to the LQ group. Apoptosis, p53 signaling, and cAMP signaling were among the top pathways targeted by the elevated miRNAs in the HQ group while oocyte meiosis, gap junction, and TGF-beta signaling were among the top pathways targeted by the elevated miRNAs in the LQ group. The supplementation of small-EVs during maturation does not affect the oocyte developmental rates. However, LQ s-EVs increase the proportion of oocytes with homogeneous mitochondrial distribution and decrease the proportion of heterogeneous distribution.Conclusion: Our findings indicated that FF-EVs contain different miRNA cargos associated with oocyte quality and could affect the mitochondrial distribution patterns during oocyte maturation.