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Gene expression analysis of in vivo originated bovine embryos fllushed out at 2- or 8-cell stages and cultured in vitro till blastocyst stage

ABSTRACT: There is an inevitable need for combining in vitro and in vivo culture systems during specific developmental time points to facilitate a comprehensive understanding of early embryo development which would yield insights into the molecular pathways controlling early development and to improve our knowledge in regulation of embryonic development. In current study, we aimed to understand the influences of in vitro culture conditions on bovine blastocyst originated in vivo at 2- or 8-cell stagre. The effect of this altrnative cuture conditions on developmental rate, gene expression pattern and subsequent influences on pathways and biological functions controlling bovine embryo development has been analyzed.

ORGANISM(S): Bos taurus

PROVIDER: GSE111990 | GEO | 2018/10/31

REPOSITORIES: GEO

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Project description:Despite many improvements with in vitro culture systems, the quality and developmental ability of mammalian embryos produced in vitro is still lower than their in vivo counterparts. To bring knowledge to answer this question, we used a mass spectrometry (MS) approach to compare the protein content of bovine embryos that were conceived in vivo or produced in vitro. A total of 38 pools of grade-1 quality bovine embryos at the 4-6 cell, 8-12 cell, morula, compact morula and blastocyst stages developed either in vivo or in vitro were analyzed by nano-liquid chromatography coupled with label-free quantitative mass spectrometry, allowing the identification of 3,028 proteins. Multivariate analysis of quantified proteins showed a clear separation of embryo pools according to their in vivo or in vitro origin at all stages. Three clusters of differentially abundant proteins (DAPs) according to embryo origin were evidenced, including 463 proteins more abundant in vivo than in vitro across development, and 314 and 222 proteins more abundant in vitro than in vivo before and after the morula stage, respectively. The functional analysis of proteins found more abundant in vivo showed an enrichment in carbohydrate metabolism and cytoplasmic cellular components. Proteins found more abundant in vitro before the morula stage were mostly localized in mitochondrial matrix and involved in ATP-dependent activity while those overabundant after morula stage were mostly localized in the ribonucleoprotein complex and involved in protein synthesis. Oviductin and other proteins previously shown to interact with early embryos in vitro were among the most overabundant proteins after in vivo development. In conclusion, the maternal environment led to faster degradation of mitochondrial proteins at early embryo developmental stages, lower abundance of proteins involved in protein synthesis at the time of embryonic genome activation and a global upregulation of carbohydrate and small molecule metabolic pathways compared to in vitro produced embryos. Furthermore, our data confirm that embryos developed in vivo uptake large amounts of oviduct fluid-derived proteins as soon as the 4-6 cell stage. These data provide new insight into the molecular contribution of the mother to the developmental ability of early embryos and will help designing better in vitro culture systems.

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Gene expression analysis of in vivo originated bovine embryos fllushed out at 2- or 8-cell stages and cultured in vitro till blastocyst stage

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