Evaluating the Development and the Expression of Bax, Bcl-2, and ErbB4 Genes following Vitrification of Eight Cell and Blastocyst Embryos

Abstract

Background & Objective: Vitrification is an effective mean for preserving embryos. During this process, expression of implanting and apoptotic genes changes and can affect the quality of embryos. In this study, the expressions of Bax and Bcl-2 as apoptotic genes and ErbB4 as implanting gene were investigated in vitrified and warmed embryos. Materials & Methods: In this study, six-eight week old female (NMRI) mice were used. Their ovaries were hyper stimulated with 7.5 IU PMSG and 7.5 IU HCG. Females were mated with males from the same strain and inspected for the presence of vaginal plugs on the following morning. Females with the presence of vaginal plugs were considered to be pregnant and were killed 62 h post HCG injection. 8-cells embryos were flushed from oviduct and were divided to three experimental groups including fresh, vitrified eight-cell embryos and vitrified blastocyst embryos. Finally, RNA was extracted and expression of Bax, Bcl-2, and ErbB4 genes was evaluated by Real time PCR. Data were analyzed by Chi-square and ANOVA tests. Results: The results of this study showed that there was not any significant difference in blastocyst formation and degeneration between three experimental groups. In addition, there was not any significant difference in relative gene expression of Bax, Bcl-2, andErbB4 between all the three groups. Conclusion: According to the result of this study, vitrification-thawed process does not have any unpleasant effect on the development of eight cell and blastocyst embryos. It does not have negative effect on Bax, Bcl2, and ErbB4 genes expression.

Keywords

• Vitrification
• Blastocyst
• Implantation