Upregulation of Toll-Like Receptor 4 Through Anti-Mir-Let-7A Enhances Blastocyst Attachment to Endometrial Cells in Mice Publisher Pubmed



Hosseini S¹ ; Hosseini S¹ ; Salehi M^{1, 3} Show Affiliations
Source: Journal of Cellular Physiology Published:2020

Abstract

Despite encouraging advances in fertility technology, the success rate of an ongoing pregnancy is relatively low and predominantly associated with implantation failure. Inflammatory responses are beneficial in the fetomaternal interface and supposedly accelerate the chances for successful implantation. The current study aims to determine the effect of Toll-like receptor 4 (TLR4) overexpression in mouse blastocysts via Let-7a downregulation using intracytoplasmic sperm injection-sperm-mediated gene transfer on embryo attachment rate. The pLenti-III-GFP-miR-Off-Let-7a vector was transmitted to oocytes derived via in vitro maturation (IVM) and in vivo oocytes by using NaOH-treated spermatozoa. Let-7a and TLR4 expression levels were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR), immunocytochemistry, and western blot analysis in both oocytes and embryos. Blastocyst adhesion on the endometrial cells was monitored by microscopic analysis. qRT-PCR results showed that Let-7a expression decreased in the IVM (GV-MII) oocytes compared to the in vivo oocyte (MII) group (p <.05). TLR4 showed a higher expression in GV-MII oocytes at both the gene and protein levels (p <.05). Following anti-miR-Let-7a transmission, the TLR4 expression level was significantly upregulated in embryos compared with the control groups (p <.05). Attachment and migration of trophoblasts cells towards endometrial cells dramatically increased compared to the control group (p <.05). Based on our results, we concluded that Let-7a might mediate embryo attachment through regulation of TLR4 expression levels. © 2020 Wiley Periodicals LLC