Division of Animal and Dairy Sciences, College of Agriculture and Life Sciences, Chungnam National University, Daejeon, Republic of Korea.
Disease Model Research Laboratory, Genome Editing Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea.
Mol Reprod Dev. 2019 Nov;86(11):1628-1638. doi: 10.1002/mrd.23250. Epub 2019 Aug 2.
Understanding preimplantation embryo development has important implications for assisted reproductive technologies (ARTs) after the introduction of in vitro fertilisation and embryo transfer because most embryonic losses occur during pre/peri-implantation. Recent studies have shown that tight junctions (TJs) are important components for embryos to develop to the blastocyst stage. However, their biological function after cavitation has not been extensively studied. We examined TJ assembly focusing on coxsackievirus and adenovirus receptor (Cxadr) and A disintegrin and metalloproteinase 10 (Adam10) using siRNA and/or an Adam10-specific inhibitor (GI254023X). TJ-associated genes, including occludin and tight junction protein 1 (Tjp1), were downregulated in the Cxadr knockdown (KD) embryos but were unaltered in Adam10 KD embryos. However, Adam10 KD or chemical inhibition affected subcellular localisation of Adam10, Cxadr, and Tjp1, leading to disrupted TJ assembly. Furthermore, Cxadr KD or GI254023X-treated blastocysts showed a relatively smaller outgrowth area and aberrant expression of transcription factor AP-2γ, a trophoblast-specific marker in the in vitro embryo outgrowth assay. In summary, we demonstrated that the Cxadr-Adam10 complex might moderate TJ integrity/stability and play pivotal roles during early embryonic development. Collectively, understanding the establishment of the TJ complex and its integrity will provide insight into translational research for predicting and selecting developmental competency for ART.
了解胚胎植入前的发育对于体外受精和胚胎移植后辅助生殖技术（ART）具有重要意义，因为大多数胚胎丢失发生在植入前/植入期。最近的研究表明，紧密连接（TJ）是胚胎发育到囊胚阶段的重要组成部分。然而，它们在空化后的生物学功能尚未得到广泛研究。我们使用 siRNA 和/或 Adam10 特异性抑制剂（GI254023X）检查 TJ 组装，重点关注柯萨奇病毒和腺病毒受体（Cxadr）和解整合素和金属蛋白酶 10（Adam10）。TJ 相关基因，包括闭合蛋白和紧密连接蛋白 1（Tjp1），在 Cxadr 敲低（KD）胚胎中下调，但在 Adam10 KD 胚胎中未改变。然而，Adam10 KD 或化学抑制会影响 Adam10、Cxadr 和 Tjp1 的亚细胞定位，导致 TJ 组装中断。此外，Cxadr KD 或 GI254023X 处理的囊胚在体外胚胎生长试验中表现出相对较小的生长面积和转录因子 AP-2γ的异常表达，AP-2γ是滋养层特异性标记物。总之，我们证明了 Cxadr-Adam10 复合物可能调节 TJ 的完整性/稳定性，并在早期胚胎发育中发挥关键作用。总之，了解 TJ 复合物的建立及其完整性将为预测和选择 ART 的发育能力提供转化研究的见解。
