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 qual-ity, 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 NovaSeq 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 develop- mental 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.

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

Institute of Animal Physiology and Genetics,Czech Academy of Sciences, 27721 Liběchov,Czech Republic;Department of Animal Production,Faculty of Agriculture,Cairo University,Giza 12613,Egypt;Department of Botany and Genetics,Faculty of Natural Sciences,Constantine the Philosopher Univer-sity in Nitra,94901 Nitra,Slovakia

畜牧与生物技术杂志（英文版）

[1]Ahmed Gad;Matej Murin;Alexandra Bartkova;Veronika Kinterova;Katerina Marcollova;Jozef Laurincik;Radek Prochazka-.Small-extracellular vesicles andtheir microRNA cargo fromporcine follicular fluids: thepotential association withoocyte quality)[J].畜牧与生物技术杂志（英文版）,2022(06):1727-1742

andtheir,fromporcine,thepotential,withoocyte,FFs,aspirated,lissamine,unstained

Small,extracellular,vesicles,follicular,fluids,association,quality,Background,Ovarian,contain,several,kinds,regulatory,factors,that,maintain,suitable,environment,development,Extracellular,EVs,among,play,essential,roles,regulating,follicle,through,molecules,include,microRNAs,miRNAs,This,study,aimed,investigate,small,Methods,Individual,oocytes,were,green,LB,vital,each,was,classified,high,HQ,low,LQ,corresponding,pooled,together,into,groups,isolated,characterized,identified,using,Illumina,NovaSeq,sequencing,platform,Additionally,utilized,effect,after,incubation,during,vitro,maturation,Results,total,including,125b,193a,5p,significantly,upregulated,while,downregulated,compared,Apoptosis,p53,signaling,cAMP,top,pathways,targeted,by,elevated,meiosis,gap,junction,TGF,beta,supplementation,does,not,affect,mental,rates,However,increase,proportion,homogeneous,mitochondrial,distribution,decrease,heterogeneous,Conclusion,Our,findings,indicated,different,cargos,associated,could,patterns

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