Abstract
In vitro maturation (IVM) and vitrification have been widely used to prepare oocytes before fertilization; however, potential effects of these procedures, such as expression profile changes, are poorly understood. In this study, mouse oocytes were divided into four groups and subjected to combinations of in vitro maturation and/or vitrification treatments. RNA-seq and in silico pathway analysis were used to identify differentially expressed genes (DEGs) that may be involved in oocyte viability after in vitro maturation and/or vitrification. Our results showed that 1) 69 genes were differentially expressed after IVM, 66 of which were up-regulated. Atp5e and Atp5o were enriched in the most significant gene ontology term “mitochondrial membrane part”; thus, these genes may be promising candidate biomarkers for oocyte viability after IVM. 2) The influence of vitrification on the transcriptome of oocytes was negligible, as no DEGs were found between vitrified and fresh oocytes. 3) The MII stage is more suitable for oocyte vitrification with respect to the transcriptome. This study provides a valuable new theoretical basis to further improve the efficiency of in vitro maturation and/or oocyte vitrification.
Highlights
In vitro maturation (IVM) of mammalian oocytes is an essential technique used in research of developmental biology and assisted reproductive technology (ART)[1]
The following four groups of mouse MII stage oocytes were used: 1. fresh germinal vesical (GV) stage oocytes matured in vitro (FG); 2. vitrified GV stage oocytes matured in vitro (VG); 3. fresh MII stage oocytes matured in vivo (FM); 4. vitrified MII stage oocytes matured in vivo (VM) (Fig. 1a)
The ATP synthase genes Atp5e and Atp5o were in the key position of the Protein-protein interaction (PPI) network (Fig. 5b). These results indicated that the mitochondrial membrane protein gene expression was different between in vitro maturation of fresh and vitrified GV stage oocytes when compared to FM
Summary
In vitro maturation (IVM) of mammalian oocytes is an essential technique used in research of developmental biology and assisted reproductive technology (ART)[1]. One major problem of MII stage oocytes is the sensitivity of the spindles to low temperatures and cryoprotective agents[13,14,15] This damage can be avoided by vitrification of the germinal vesical (GV) stage oocytes, a stage when the spindle apparatus has not yet formed. RNA-seq was used to analyse the transcriptomes of differently treated mouse MII stage oocytes to gain a better understanding of the transcriptome events in the oocytes after in vitro maturation and/or vitrification. This study provided a transcriptional basis for using oocyte vitrification to preserve female fertility for those who may lose their ovarian function because of surgery, cancer treatment and premature menopause or who decide to postpone having children
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