Whole Transcriptome Sequencing And Analysis Of Bovine Blastocysts Produced From In Vivo And In Vitro

In vitro embryo production technology has been widely used as an efficient and simple assisted reproductive technology.The production of embryos in vitro is regarded as the third revolution in livestock breeding.In vitro fertilization technology is an important part of in vitro embryo production technology.It is of great significance to improve the breeding efficiency of livestock and poultry and improve the genetics of excellent livestock.Vitrification is popular because of low cost,time consumption,simple operation and high efficiency.However,compared with in vivo embryos,in vitro embryo production still has deficiencies in cleavage rate,pregnancy rate,calving rate,etc.These may be related to abnormal transcriptome(including mRNA,lncRNA,circRNA).In this study,single-cell whole transcriptome sequencing technology was used to analyze the whole transcriptome of in vivo bovine blastocysts,bovine blastocysts from IVF of fresh oocytes,and bovine blastocysts from IVF of vitrified oocytes(including mRNA,lncRNA,circRNA)and the results of GO annotation analysis and KEGG enrichment analysis of differentially expressed genes.The main findings of this experiment are as follows:1.The whole transcriptome sequencing of in vivo and in vitro bovine blastocysts was used to explore the mRNA pattern and analyze the biological information.There were 78 differentially expressed genes between in vivo and IVF groups,60 genes were up-regulated and 18 were down-regulated in IVF group.There were 362 differentially expressed genes between In vivo group and V_IVF group,of which 305 were up-regulated,57 down-regulated in V_IVF group;There were 14 differentially expressed genes between IVF and V_IVF groups,of which 13 were up-regulated and 1 down-regulated V_IVF group.GO annotation of its differentially expressed genes found that differentially expressed genes in vivo and in vitro blastocysts were enriched in entries such as rRNA processing,metabolic processes of organic nitrogen compounds,biosynthetic processes of organic nitrogen compounds.Through KEGG Functional enrichment analysis found that differentially expressed genes were significantly enriched in glycolysis/gluconeogenesis pathways,and 29 genes were significantly up-regulated in this pathway.2.The whole transcriptome sequencing of in vivo and in vitro bovine embryos was used to discuss the lncRNA pattern and analyze the biological information.There were 187 differentially expressed lncRNAs between in vivo and IVF group,up-regulated expression lncRNA were 124,down-regulated expression lncRNA were 63 in IVF group.There were 694 differentially expressed lncRNAs between in vivo and V_IVF group,up-regulated expression lncRNA were 223,down-regulated expression lncRNA were 471 in V_IVF group.There were 114 differentially expressed lncRNA between IVF and V_IVF groups,up-regulated expression lncRNA were 27,down-regulated expression lncRNA were 87 in V_IVF group.GO annotation of the differentially expressed lncRNA found that they were enriched into the items such as cell growth and development,cytoskeleton,ATP binding.Differentially expressed lncRNA were found through KEGG functional enrichment analysis in lysosomes,RNA transport,and oxidative phosphorylation pathways,oxidative phosphorylation pathways are essential for early embryonic development and successful pregnancy outcomes.3.The transcriptome sequencing of bovine embryos produced in vivo and in vitro was used to explore the circRNA pattern and analyze the biological information,and 994 circleRNAs were obtained.These circleRNAs have 437 target genes.Differentially expressed circleRNA between IVF and in vivo groups were 28,of which 2 circleRNA expressions increased and 26 expressions decreased in IVF group.Differentially expressed circleRNA between V_IVF and in vivo groups were 51,which included 2 expressions increased and 49 expressions decreased in V_IVF group.Differentially expressed circleRNA between IVF and V_IVF groups were 1.GO annotation of Differentially expressed circleRNA found that these circleRNA were enriched in the positive regulation of chromosomal organization,the regulation of histone H3-K9 methylation and other pathways.