Abstract
The first segregation at the blastocyst stage is the symmetry-breaking event to characterize two cell components; namely, inner cell mass (ICM) and trophectoderm (TE). TEA domain transcription factor 4 (TEAD4) is a well-known regulator to determine TE properties of blastomeres in rodent models. However, the roles of bovine TEAD4 in blastocyst development have been unclear. We here aimed to clarify the mechanisms underlining TE characterization by TEAD4 in bovine blastocysts. We first found that the TEAD4 mRNA expression level was greater in TE than in ICM, which was further supported by TEAD4 immunofluorescent staining. Subsequently, we examined the expression patterns of TE-expressed genes; CDX2, GATA2 and CCN2, in the TEAD4-knockdown (KD) blastocysts. These expression levels significantly decreased in the TEAD4 KD blastocysts compared with controls. Of these downregulated genes, the CCN2 expression level decreased the most. We further analyzed the expression levels of TE-expressed genes; CDX2, GATA2 and TEAD4 in the CCN2 KD blastocysts. Strikingly, the CCN2 KD blastocysts showed the downregulation of CDX2, GATA2 and TEAD4 Furthermore, the ratio of TE-to-ICM cell numbers in the CCN2 KD blastocysts significantly decreased compared to controls. To our knowledge, this is the first study showing the regulation of CCN2 expression thorough TEAD4 in mammalian embryos. Not only that, this study also provides evidence that reciprocal regulation of TEAD4 and CCN2 is required for TE development with appropriate gene expression in bovine blastocysts.