Specific activation of embryonic IFNAR1 and endometrial IFNAR2 induced by embryonic IFNτ directs normal uterine fate for bovine early implantation

Background: Interferon-tau (IFNτ), as an antiluteolytic factor secreted by trophoderm during the pregnancy of ruminants, actually functions by activating the IFNτ receptor 1 (IFNAR1) and IFNτ receptor 2 (IFNAR2). However, it has not been clearly understood how IFNτ-IFNAR cascade regulation processes between the embryo and uterine epithelial cells in ruminants. Methods: CRISPR/Cas9 technology was used to compare the differences of IFNτ secretion in In vitro fertilization(IVF), parthenogenetic activation(PA) and somatic cell nuclear transfer (SCNT) and their in vitro co-culture effect with uterine endometrial epithelial cell with those in vivo ones in this study.Results: In this study, we found the expression and location of IFNτ in the blastocysts from different sources. IFNτ, IFNAR1 and IFNAR2 were all located in the trophoblast cells of the blastocyst. However, the fluorescence intensity of IFNAR1 was consistent with that of IFNτ. Antagonizing the expressions of IFNAR1 and IFNAR2 in embryos and co-culture with EEC reduced the expressions of Integrin αv β3, WNT7A, and ISG15 in EECs. Knocking out IFNAR1 and IFNAR2 reduce the expressions of Integrin αv β3 and WNT7A in EECs, the deletion of IFNAR2 gene has a greater impact than that of IFNAR1 gene. IFNAR1-/IFNAR2+ and IFNAR1+/IFNAR2- EECs were co-cultured with IVF embryos, the expression of Integrin αv β3 was inhibited, and the inhibition of IFNAR1+/IFNAR2- was much stronger, and the expression of WNT7A was not inhibited. The expressions of Integrin αv β3 and WNT7A did not change significantly after IFNAR1-/IFNAR2+ and IFNAR1+/IFNAR2- co-culture with PA embryos. Conclusions: All of these results strongly suggest that specific activation of embryonic IFNAR1 and endometrial IFNAR2 induced by embryonic IFNτ directs normal uterine preparation for bovine early implantation.