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Loss- and Gain-of-function Approach to Investigate Early Cell Fate Determinants in Preimplantation Mouse Embryos
Published on: June 6, 2016
MicroRNA-223-3p suppresses leukemia inhibitory factor expression and pinopodes formation during embryo implantation
Xiyuan Dong1, Cong Sui1, Kai Huang1
1Reproductive Medicine Center, Tongji Hospital, Tongji Medicine College, Huazhong University of Science and Technology Jiefang Avenue 1095#, Wuhan 430030, People's Republic of China.
Abstract:
MicroRNA (miRNA) regulates gene expression in a post-transcriptional manner, which hybridizes the target mRNAs with complementary sequence and subsequently leads to translation repression or mRNA degradation. Online sequence alignment showed that there is a putative binding site of miR-223-3p on the 3'UTR of LIF, which is considered to be an important marker of endometrial receptivity. Thus, we hypothesized that miR-223-3p may affect embryo implantation by suppressing LIF expression. In this study, we found that miR-223-3p and LIF protein was inversely expressed in the endometrium of mice during implantation window. Then we proved that miR-223-3p directly binds to LIF 3'UTR with luciferase reporter assay and supresses the expression of LIF. To investigate whether miR-233-3p affects embryo implantation, miR-223-3p agonist was injected into the uteri of pregnant mice. The results demonstrated the suppressing effect of miR-223-3p on embryo implantation. Furthermore, over-expression of miR-223-3p was found to compromise pinopodes formation in the endometrium of mice. Taken all together, our findings revealed that miR-223-3p suppresses pinopodes formation and LIF protein expression, which may lead to diminished embryo implantation.
Insights
MicroRNA-223-3p suppresses embryo implantation by reducing LIF protein and inhibiting uterine pinopodes formation. This microRNA (miRNA) plays a key role in regulating gene expression during early pregnancy.
Area of Science:
- Reproductive Biology
- Molecular Endocrinology
- Gene Regulation
Background:
- MicroRNAs (miRNAs) are key post-transcriptional regulators of gene expression.
- Leukemia inhibitory factor (LIF) is crucial for endometrial receptivity and embryo implantation.
- A potential interaction between miR-223-3p and LIF suggests a role in implantation.
Purpose of the Study:
- To investigate the role of miR-223-3p in regulating LIF expression and its impact on embryo implantation.
- To elucidate the mechanism by which miR-223-3p influences endometrial receptivity.
Main Methods:
- Bioinformatic analysis to identify miR-223-3p binding site on LIF 3'UTR.
- Luciferase reporter assay to confirm direct binding and suppression.
- In vivo studies involving uterine injection of miR-223-3p agonist in pregnant mice.
- Histological examination of endometrial pinopodes formation.
Main Results:
- Inverse expression of miR-223-3p and LIF protein in the mouse endometrium during the implantation window.
- miR-223-3p directly binds to the LIF 3'UTR and suppresses LIF expression.
- In vivo administration of miR-223-3p agonist significantly inhibited embryo implantation.
- Overexpression of miR-223-3p compromised the formation of endometrial pinopodes.
Conclusions:
- miR-223-3p negatively regulates LIF protein expression.
- miR-223-3p impairs embryo implantation by suppressing LIF and compromising pinopodes formation.
- These findings highlight miR-223-3p as a potential regulator of uterine receptivity.
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