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Toll-Like Receptor4 As a Modulator of Fertilization and Subsequent Pre-Implantation Development Following in Vitro Maturation in Mice Publisher Pubmed



Hosseini S1, 2 ; Dehghanimohammadabadi M2 ; Ghafarri Novin M1 ; Haji Molla Hoseini M3 ; Arefian E4 ; Mohammadi Yeganeh S5 ; Salehi M5, 6
Authors
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Authors Affiliations
  1. 1. Department of Reproductive Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  2. 2. Department of Transgenic Animal Science, Stem Cell Technology Research Center, Tehran, Iran
  3. 3. Department of Immunology, Faculty of Medicine, Shahid Beheshti University of Medical Science, Tehran, Iran
  4. 4. Department of Microbiology, School of Biology, College of Science University of Tehran, Tehran, Iran
  5. 5. Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  6. 6. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Source: American Journal of Reproductive Immunology Published:2017


Abstract

Problem: The use of in vitro maturation (IVM) as an alternative approach to the conventional assisted reproductive technique in clinical practice is limited due to low fertilization rate. The expression of Toll-like receptors (TLRs) as members of an innate immune system in cumulus cells are thought to affect fertilization. This study was aimed to evaluate the effect of IVM on TLR4 gene expression and fertilization rate in oocytes derived from IVM in comparison with in vivo matured one. Method of study: Cumulus cells are collected from oocytes derived IVM and in vivo. The expression level of Tlr4 in cumulus cells of both groups was assessed by quantitative real-time PCR. To examine the protein expression level of TLR4, immunocytochemistry and Western blotting techniques were carried out. TLR4 receptor functions were also confirmed by blockade assay and TLR4 receptor activation with lipopolysaccharide. Result: There was a substantial decrease in fertilization and blastulation rate in the IVM group in comparison with the in vivo one. The mRNA expression and protein levels of TLR4 declined in cumulus cells following IVM. Anti-TLR4 blocking antibody dramatically decreased the rate of fertilization and blastocyst formation compared to the in vivo group. In contrast, the fertilization rate was enhanced significantly in the presence of LPS as a TLR4 ligand compared to the control group. Conclusion: Low expression level of Tlr4 following IVM and higher fertilization rate through TLR4 receptor activation with LPS proposed that alteration in TLR4 expression and subsequent cytokine section could be a possible cause of low fertilization rate in IVM-derived oocytes. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd