| Embry implantation refers to the process that the embryo contact with maternal uterus and establishes a structural bond,and finally realizes the maternal-fetal material exchange,which mainly includes the establishment of endometrial receptivity(ER)and the stages of embryo development,migration,localization,adhesion and invasion.This process regulated by complex and precise regulation is a research hotspot.The synchrony of the uterus and embryo development determines whether implantation can be successful.Goat is ruminants which is surface implantation.However,the mechanism of goat embryo implantation during early pregnancy is still not clear at present,and there is not much work on the excavation of genes related embryo implantation at uterus.Since transcriptome sequencing(RNA-Seq)can quickly and comprehensively obtain transcription products from tissues or cells under different conditions,it is possible to use it to explore genes related embryo implantation at uterus.Therefore,Our study took Dazu Black Goat,a national genetic resource,as the research object.And we analyzed the changes of uterine histomorphology during different gestation periods,and screened the genes related embryo implantation at uterus during early pregnancy by transcriptomics.The purpose of our study is to providing a preliminary basis for the analysis of the molecular mechanism regulating goat embryo implantation.Our study collected uterine tissue in non-gestation(KH),pregnancy 15day(P15)and pregnancy 19 day(P19)of Dazu Black Goat.And we analyzed the uterus tissue morphology during early pregnancy by H&E staining and immunohistochemistry.Then,RNA-Seq was used to screen the key regulatory genes affecting goat embryo implantation,and RT-q PCR was used to verify the sequencing results.The main research results were as follows:1.Morphological analysis: Compared with KH,the endometrial thickness of the uterine horn increased,the folds increased,the uterine glands increased and became larger in P15 and P19.PCNA(Proliferating Cell Nuclear Antigen)detected in luminal epithelium cells(LE),glandular epithelial cells(GE)and stromal cells(SC)at different gestation.The expression of PCNA in LE and GE was higher than that of SC at same period.2.RNA-Seq analysis: There were 48 differentially expressed genes in the P15 group compared with the KH group,of which 12 were up-regulated and 36 were down-regulated.By GO analysis,it was found that the differentially expressed genes were mainly related to extracellular matrix,organelle membrane,mitochondrial composition,ferritin receptor activity,collagen fibrillary tissue,etc.KEGG analysis showed that differential genes were mainly involved in cell adhesion molecules,metabolism,antigen processing and expression,thermogenesis and other pathways.There were 1439 differentially expressed genes in the P19 group compared with the P15 group,among which 929 genes were up-regulated and 510 genes were down-regulated.GO analysis showed that the differentially expressed genes were mainly related to immune response,inflammatory response,protein binding,immunoglobulin receptor activity,cell localization,cell surface,etc.KEGG analysis showed that the differential genes were mainly concentrated in metabolic pathways,cytokine receptor interactions,Toll-like receptor signaling pathways,TNF signaling pathways,cell adhesion molecules and other pathways.3.Finally,a total of 8 differentially expressed genes(C1S,CSF1,CST6,SLC24A4,HOXA10,HOXA11,MMP9 and ITGA11)were selected and verified by RT-q PCR,and the verification results were consistent with the sequencing results.In conclusion,the differentially expressed genes related to embryo implantation in the uterus of Dazu Black Goat during early pregnancy were screened in this study and providing a preliminary basis for studying the molecular mechanism of goat embryo implantation. |
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