| Vangl2first discovered and identified was the vertebrate homolog ofVan Gogh (Vang) gene in Drosophila, and its serine cluster motif in theamino-terminal cytoplasm and the PDZ motif in the carboxy-terminalcytoplasm domain make it very conservative in the evolution. The gene hashomologous genes Vangl1in human, mouse, zebrafish, and they share73.1%homology, this structural homology also determines the similarity oftheir functional. Existing data show that the loss of Vangl1and Vangl2function can lead to abnormal embryonic development and introduce thedeath of the embryo during pregnancy in severe cases. In fact, due to itsinterference with the Wnt/PCP signal transduction pathway in the gastrulaand neurula developmental processes, affecting cell migration, adhesion,polarity, and the remodeling of the cytoskeleton. Embryo implantation wasas the primary key in the whole process of pregnancy, the success or notdirectly affects the embryo development and pregnancy outcome. However,whether Vangl1and Vangl2play a role during embryo implantationremains unclear. Therefore, this research mainly explores the expression of Vangl1and Vangl2in the uterus of mice during peri-implantation and itsinfluence on embryo implantation.The expression of of Vangl1and Vangl2was examined by real-timePCR, in situ hybridization, immunohistochemistry and western blot in thenormal pregnant mice and pseudo-pregnant mouse uterine tissue. Thisexperiment was performed by uterine angle injection with antisenseoligodeoxynucleotides (A-ODNS) of Vangl1and Vangl2on day3.5,remove endometrial tissue after24hours, then respectively detect theexpression differences of Vangl1and Vangl2mRNA and protein andobserve the embryo number and morphology changes on day8afteroperation. The results of this study were as follows:1. The results of Real-time PCR showed that that the Vangl1and Vangl2mRNA levels was significantly upgraded with the increase of gestationaldays and reached maximum levels on day5. The expression of the Vangl1mRNA and protein in the uterus of pseudo-pregnant mice was significantlylower than that of pregnant mice on D4-D6（*P<0.05）. The Vangl2mRNAand protein had no quantitative change between pregnancy andpseudo-pregnancy mice.2. The results of in situ hybridization showed Vangl1mRNA was mainlylocalized in the stromal cells, the luminal and glandular epithelium, anddecidual cells is also highly expressed on d6of pregnancy. Vangl2mRNAwas mainly localized in the luminal, glandular epithelium and decidual cells. MRNA positioning of these two genes in the pseudo-pregnant groupwere similar with the normal group.3. Western blot results and Real-time PCR results were consistent.4. Immunohistochemistry analysis showed that the Vangl1protein mainlylocated in the stromal cells and decidual cells, and the Vangl2protein washigher expressed in the luminal and the glandular epithelium on D5andlower expressed in stromal cells on D3. The results of Western blot andimmunohistochemical were basically identical. Protein positioning of thesetwo genes in the pseudo-pregnant group were similar with the normalgroup.5. Vangl1and Vangl2mRNA and protein expression were inhibited byuterine angle injection, and the number of embryo implantation in micewas significantly reduced（*P<0.05）.In summary, our results indicate that Vangl1and Vangl2havetime-dependent up-regulated during the embryo implantation period. Thisfinding suggests that they might be essential for embryo implantation, andthe expression pattern of Vangl1gene induced embryo source signal wasmore sensitive compared with Vangl2. Uterine angle injection showsVangl1and Vangl2may play an important role in the process of embryoimplantation. However, their expression positioning in the uterus weredifferent, this showed Vangl1and Vangl2might perform differentfunctions during embryo implantation. |
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