Comparative Transcriptome Analysis During Early Development Between Three Different Types Embryos Of Mongolia Sheep

Mongolia sheep are special local livestock which have excellent meat quality.However,due to the special geographical and climatic conditions,their reproductive performance is generally low.It can preserve and expand high-quality sheep germplasm resources effectively and shorten the genetic breeding cycle by using in vitro fertilization and somatic cell nuclear transfer technology to produce embryos.Still now,in vitro embryos produced by existing technologies not only have low common development rate problem,but also have obvious developmental block.To explore the mechanism of the potential difference between in vitro fertilization and somatic cell nuclear transfer embryos,firstly we chose three different types and stages of early embryonic development time(2-cell,4-cell and 8-cell)and extract the mRNA of every single embryo.Secondly,we successfully established the cDNA library by reverse transcription of the samples,and complete the sequencing and construction of the above embryo transcriptome by using the Illumina HiSeqX10 high-throughput sequencing system.Lastly,the differences in gene expression of different types embryos were analyzed.The results we analysised are shown below:(1)Successfully amplified and established the cDNA library of the early embryo of Mongolian sheep from different sources and construct the transcriptome by using single cell RNA extraction system Smart-seq V4;(2)There are 530 significant different genes shows in gene expression between in vivo fertilization embryos from 2-cell to 4-cell while 869 shows between 4-cell to 8-cell;4550 significant different genes shows in gene expression between in vitro fertilization embryos from 2-cell to 4-cell while 4687 shows between 4-cell to 8-cell;4600 significant different genes shows between somatic cell nuclear transfer embryos from 2-cell to 4-cell while 5212 shows between 4-cell to 8-cell;(3)Through GO enrichment analysis we found that the differentially expressed genes in in vivo fertilization embryos is mainly expressed in cell metabolism,nucleic acid metabolism,macromolecular metabolism and cellular component organization or biogenesis.Compared to the in vivo fertilization embryos,the difference gene of other two kinds embryos expressed in metabolic process,cellular development process and annular macromolecular metabolism,nitrogen metabolism;(4)Through KEGG analysis,the expression of differentially expressed genes in these three different source embryos was mainly distributed in the cell pathway such as ATP-binding cassette transporters,Oxidative phosphorylation,cAMP signaling pathway,Cell adhesion molecules(CAMs),Mismatch repair and Phagosome.