The Cxadr–Adam10 complex plays pivotal roles in tight junction integrity and early trophoblast development in mice

Abstract

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.

Tight Junction Assembly Ensures Maintenance of Pregnancy during Embryogenesis in a Mouse Model

“…In agreement with a previous study (Jeong et al, 2019), Cxadr KD resulted in retardation of blastocyst (52 ± 3%, Fig. 1A), and morula embryos treated with selective inhibition (SI) of Adam 10 also arrested and developed blastocyst (45 ± 6%, Fig.…”

“…The purpose of this study is to investigate the effect of defects of TJ complex/integrity on postimplantation embryo development and to expand our previous findings (Kwon et al, 2016a;Kwon et al, 2016b;Jeong et al, 2019) where CXADR interacts with ADAM 10 and they are required for AJ/TJ assembly during preimplantation development in pig and mouse. In line with previous studies (Kwon et al, 2016a;Kwon et al, 2016b;Jeong et al, 2019), we found that depletion of Cxadr and inhibition of 2006). Thus, the gradual weight gain after ET may reflect increased embryo and placental weight, but weight-loss for two consecutive days is attributed to loss of embryos because maternal body weight rapidly increased from around 2nd trimester onwards (Kulandavelu et al, 2006).…”

“…To explore potential use of TJ complex integrity as an embryo mortality related bio-marker, we expand our previous finding where CXADR is involved in AJ/TJ formation and ADAM10 interacts with CXADR and TJP1 for is sequential TJ assembly during preimplantation development (Jeong et al, 2019). We first examined preimplantation developmental competency by deletion of components of TJ complex injecting Cxadr (Coxsackie virus and adenovirus receptor) siRNA or using disruption/ inhibition of TJ complex using a chemical inhibitor of Adam 10 (GI254023X).…”

Proteomic analysis of extracellular vesicles secreted by primary human epithelial endometrial cells reveals key proteins related to embryo implantation

“…Proteins in this group included ADAM10, CPM, GPX3, and UBA52. ADAM10 is a metalloproteinase that may moderate tight junctions in trophoblastic cells, which is an essential component for the development of the embryo to the blastocyst stage [ 54 ]. CPM is a cell surface peptidase expressed in trophoblastic cells, which is involved in its differentiation and function [ 55 ].…”

Transcriptomic signatures in trophectoderm and inner cell mass of human blastocysts with different expectations of pregnancy

“…The number of DEGs in the other groups was too small to perform GO analysis. Notably, among 11 DEGs whose expression levels were higher in the TE of blastocysts with higher expectations of pregnancy, CXADR, CLDN10, and ATP1B were known to be involved in normal development [24][25][26] (see Discussion). These results provide a valuable list of genes for predicting pregnancy expectations in blastocysts.…”