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

Genes

Jorge Jimenez-Almazán¹  Felipe Vilella¹  Blanca Simon²  David Bolumar²  Alicia Amadoz³  Diana Valbuena³  Inmaculada Moreno³ 

[1] D, Parque Tecnológico de Paterna, Ronda de Narcís Monturiol, 11B, 46980 Paterna, Spain;Igenomix Foundation-INCLIVA Biomedical Research Institute, Ronda de Narcís Monturiol, 11B, 46980 Paterna, Spain;;Igenomix, R&

关键词: extracellular vesicles;    exosomes;    microvesicles;    apoptotic bodies;    dna;    preimplantation embryos;    murine blastocysts;   

DOI  :  10.3390/genes11020203

来源: DOAJ

【 摘 要 】

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 (20−500 nm) and total concentrations of 1.74 × 10⁷ ± 2.60 × 10⁶ particles/mL. Embryo secreted EVs were positive for CD63 and ARF6. DNA cargo sequencing demonstrated no differences in DNA between apoptotic bodies or smaller EVs, although they showed significant gene enrichment compared to control medium. The analysis of sequences uniquely mapping the murine genome revealed that DNA contained in EVs showed higher representation of embryo genome than vesicle-free DNA. Murine blastocysts secrete EVs containing genome-wide sequences of DNA to the medium, reinforcing the relevance of studying these vesicles and their cargo in the preimplantation moment, where secreted DNA may help the assessment of the embryo previous to implantation.

【 授权许可】

Unknown