From follicle to blastocyst:microRNA-34c from follicular flui-derived extracellular vesicles modulates blastocyst quality  

作　　者：Camilla Benedetti Krishna Chaitanya Pavani Yannick Gansemans Nima Azari‑Dolatabad Osvaldo Bogado Pascottini Luc Peelman Rani Six Yuan Fan Xuefeng Guan Koen Deserranno Andrea Fernandez‑Montoro Joachim Hamacher Filip Van Nieuwerburgh Trudee Fair An Hendrix Katrien Smits Ann Van Soom 

机构地区：[1]Department of Internal Medicine,Reproduction and Population Medicine,Faculty of Veterinary Medicine,Ghent University,9820 Merelbeke,Belgium [2]Department for Reproductive Medicine,Ghent University Hospital,9000 Ghent,Belgium [3]Laboratory of Pharmaceutical Biotechnology,Faculty of Pharmaceutical Sciences,Ghent University,B‑9000 Ghent,Belgium [4]Department of Animal Science,University of Tennessee,Knoxville,TN,USA.5 Department of Nutrition,Genetics and Ethology,Faculty of Veterinary [5]Medicine,Ghent University,B‑9000 Ghent,Belgium [6]Institute of Crop Science and Resource Conservation,Plant Pathology,Rheinische Friedrich-Wilhelms-University of Bonn,53115 Bonn,Germany [7]School of Agriculture and Food Science,University College Dublin,Belfield,Dublin,Ireland [8]Laboratory of Experimental Cancer Research,Department of Human Structure and Repair,Ghent University,B‑9000 Ghent,Belgium

出　　处：《Journal of Animal Science and Biotechnology》2024年第6期2276-2292,共17页畜牧与生物技术杂志(英文版)

基　　金：supported by the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 860960;by Bijzonder Onderzoeksfonds GOA(Geconcerteerde onderzoeksacties)2018000504(GOA030-18 BOF);supported by the Research Foundation Flanders(FWO)(grant numbers:1228821N and 12A2H24N)。

摘　　要：Background Within the follicular fluid,extracellular vesicles(EVs)guide oocyte growth through their cargo microRNAs(miRNAs).Here,we inv estigated the role of EVs and their cargo miRNAs by linking the miRNAs found in EVs,derived from the fluid of an individual follicle,to the ability of its oocyte to become a blastocyst(competent)or not(non-competent).Methods Bovine antral follicles were dissected,categorized as small(2-4 mm)or large(5-8 mm)and the corresponding oocytes were subjected to individual maturation,fertilization and embryo culture to the blastocyst stage.Follicular fluid was pooled in 4 groups(4 replicates)based on follicle size and competence of the corresponding oocyte to produce a blastocyst.Follicular fluid-derived EVs were isolated",characterized,and subjected to miRNAsequencing(Illumina Miseq)to assess differential expression(DE)in the 4 groups.Functional validation of the effect of miR-34c on embryo development was performed by supplementation of mimics and inhibitors during in vitro maturation(IVM).Results We identified 16 DE miRNAs linked to oocyte competence when follicular size was not considered.Within the large and small follicles,46 DE miRNAs were driving blastocyst formation in each group.Comparison of EVs from competent small and large follicles revealed 90 DE miRNAs.Cell regulation,cell differentiation,cell cycle,and metabolic process regulation were the most enriched pathways targeted by the DE miRNAs from competent oocytes.We identified bta-miR-34c as the most abundant in follicular fluid containing competent oocytes.Supplementation of miR-34c mimic and inhibitor during IVM did not affect embryo development.However,blastocyst quality,as evidenced by higher cell numbers,was significantly improved following oocyte IVM in the presence of miR-34c mimics,while miR-34c inhibitors resulted in the opposite effect.Conclusion This study demonstrates the regulatory effect of miRNAs from follicular fliived EVs on oocyte competence acquisitividing a further basis for understanding the significance

关 键 词：Developmental competence Extracellular vesicles MATURATION MicroRNA miR-34c OOCYTE 

分 类 号：Q132.4[生物学—普通生物学]