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RESEARCH PRODUCT

Identification and Characterization of Extracellular Vesicles and Its DNA Cargo Secreted During Murine Embryo Development

Diana VabuenaAlicia AmadozInmaculada MorenoDavid BolumarBlanca SimonFelipe VilellaJorge Jimenez-almazán

subject

0301 basic medicinemurine blastocystslcsh:QH426-470Embryonic DevelopmentexosomesArticleEmbryo Culture Techniques03 medical and health scienceschemistry.chemical_compoundMice0302 clinical medicineGeneticsmedicineAnimalsSecretionBlastocystParticle Sizeapoptotic bodiesGenetics (clinical)030219 obstetrics & reproductive medicineChemistryADP-Ribosylation FactorsTetraspanin 30BlastocoelHigh-Throughput Nucleotide SequencingEmbryoImmunogold labellingSequence Analysis DNADNAEmbryonic stem cellMicrovesiclesCell biologylcsh:Genetics030104 developmental biologymedicine.anatomical_structureBlastocystADP-Ribosylation Factor 6Culture Media Conditionedembryonic structuresOocytespreimplantation embryosFemaleextracellular vesiclesmicrovesiclesDNA

description

Extracellular vesicles (EVs) are known to transport DNA, but their implications in embryonic implantation are unknown. The aim of this study was to investigate EVs production and secretion by preimplantation embryos and assess their DNA cargo. Murine oocytes and embryos were obtained from six- to eight-week-old females, cultured until E4.5 and analyzed using transmission electron microscopy to examine EVs production. EVs were isolated from E4.5-day conditioned media and quantified by nanoparticle tracking analysis, characterized by immunogold, and their DNA cargo sequenced. Multivesicular bodies were observed in murine oocytes and preimplantation embryos together with the secretion of EVs to the blastocoel cavity and blastocyst spent medium. Embryo-derived EVs showed variable electron-densities and sizes (20&ndash

10.3390/genes11020203http://dx.doi.org/10.3390/genes11020203