Establishment of the Adherens junction (AJ) and Tight junction (TJ) are important steps in terms of morphological formation during preimplantation develoment. Particularly, TJ complex is crucial for cavitation in blastocyst. So far, many TJ protein/genes are revealed. However, the biological function and regulation of TJ were not elucidated during post implantation. We depleted several TJ and TJ associated genes using RNA interference, and examined preimplantation development with TJ. We tested functionality of paracellular sealing to determine integrity of TJ formation and examined TE differentiation indirectly using outgrowth assay in vitro. We observed defect of paracellular permeability in the TJ related genes knockdown(KD) blastocyst and abnormal outgrowth. Particularly, trophoblast cells were not stretched out in the KD groups. Finally, we did embryo transfer using the TJ genes KD and control blastocysts into surrogate mothers. We found lower of the implantation rates/ maintenance of pregnancy in the TJ KD groups (less than 40%) than in the controls (about 80%). In conclusion, TJ integrity is can be used as a selective marker for developmentally competent embryos and successful pregnancy.

Transcription factor called activating enhancer binding protein 2C (AP2-gamma) is found in a variety of species and expressed from oocyte stage onwards, particularly restricted to the trophectoderm. Recent studies demonstrated that ablation of Tfap2c led to failure of tight junction biogenesis, particularly the knock-down embryos of Tfap2c did not form cavity from morula to blastocyst in mouse and pig. We speculated that the Tfa2pc may also be involved in desmosome biogenesis because blastocoel formation is coincident with the establishment of desmosome. To determine this, we depleted Tfap2c injecting siRNA into one-cell zygote and analysed the expression levels of genes that are required for desmosome complex such as PkP2, Pkp3, Dsc2, and Dsg2. We found only Pkp3 was up-regulated in the knockdowned morula embryos. Interestingly, upstream region of Pkp3 had putative Tfap2c binding sites. In conclusion, our results suggest that Tfap2c is not a crucial factor but somehow it might be involved in desmosome biogenesis directly or indirectly via Pkp3.