Abstract
We measured differential expression profiles of genes and long non-coding RNA (lncRNA) using RNA sequencing in bovine embryos with or without glutathione (GSH) treatment. Bovine embryos fertilized in vitro were treated with GSH to blastocyst. Embryos at the 8-16-cell and morula stages were collected, with embryos without GSH treatment as the control. RNA was isolated, amplified, and sequenced. Differentially expressed genes (DEGs) and lncRNAs (DElncRNAs) were identified and bioinformatic analyses carried out. Transcript levels were confirmed using quantitative RT-PCR. A total of 4100 DEGs were identified, of which 3952 were in GSH-treated morulae and 884 in untreated morulae. More gene ontology (GO) terms were associated with GSH treatment than with control conditions. KEGG analysis showed that glutathione metabolism, citrate cycle, and metabolic pathways involving glycine, serine, and threonine were observed only in GSH-treated embryos. Among 4273 DElncRNAs identified, 59 were potentially important in GSH-treated embryo development, including 14 involved in glutathione metabolism. The 59 DElncRNAs co-expressed with protein-coding mRNAs involved similar GO terms and pathways as the DEGs. This appears to be the first comprehensive profiling of DEGs and DElncRNAs in bovine embryos fertilized in vitro with or without GSH, and the first systematic screen of potential lncRNAs in bovine embryos.
Highlights
IntroductionMammalian pre-implantation embryonic development is a comprehensive and finely controlled process that includes fertilization, cleavage divisions, genome activation, compaction, and blastulation
Mammalian pre-implantation embryonic development is a comprehensive and finely controlled process that includes fertilization, cleavage divisions, genome activation, compaction, and blastulation.Following fertilization, the maternal-to-zygotic transition is the first critical transition in development.Following this, the developmental program is directed by the activated embryonic genome, instead of maternal proteins and mRNA [1]
In GSH-treated embryos, GSH metabolism was enriched among protein-coding mRNAs that co-expressed with 14 DElncRNAs, but this enrichment was not observed in untreated embryos (Table S17)
Summary
Mammalian pre-implantation embryonic development is a comprehensive and finely controlled process that includes fertilization, cleavage divisions, genome activation, compaction, and blastulation. Antioxidants 2020, 9, 402 the intracellular redox balance toward oxidation [6,7] This alters the cell fate determination, especially in embryos cultured or produced in vitro [8]. Adding GSH to culture medium increased the developmental rate of bovine morula and blastocyst produced by in vitro fertilization by 13% and 20%, respectively [20]. Our previous work showed that treating in vitro fertilized embryos with GSH significantly increased the rate of morula formation but not of 8-16-cell stage embryos [20]. The results obtained here with bovine embryos may provide some clues about human embryos because of the similarities in their transcriptomes [2]
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