期刊
THERIOGENOLOGY
卷 125, 期 -, 页码 317-323出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.theriogenology.2018.11.013
关键词
Buffalo; beta-mercaptoethanol; Embryo; Implantation; Apoptosis
资金
- National Key Research and Development Program [2017YFE0113800]
- Key Research and Development Program in Guangxi, China [GuiKe AB16380040]
The objectives of the present study were to investigate the effect of vitrification on the expression of the key genes associated with blastocyst developmental potential (beta-catenin-catenin, E-cadherin, Oct-4, Cdx2, Gata3), and whether the presence of beta-mercaptoethanol (beta-ME, 100 mu M) in in vitro culture (IVC) media will affect the expression of these genes. Buffalo pre-implantation embryos were divided into three groups: (1) fresh non-vitrified embryos were used as control, (2) vitrified embryos cultured with beta-ME (+), and (3) vitrified embryos cultured without (-) beta-ME. The results showed that all genes were affected by vitrification, however, the presence of beta-ME in IVC media significantly (P < 0.05) modified the expression level of beta-catenin, E-cadherin and Oct-4 in vitrified blastocyst compared to those cultured without beta-ME. Protein expression analysis by immunofluorescence and western blot also revealed that the expression level of beta-catenin and E-cadherin was significantly higher in vitrified embryos cultured with beta-ME than those cultured without beta-ME, which, in turn, was lower than fresh control group. However, there was no significant difference between vitrified groups in the expression level of Cdx2 and Gata3. Furthermore, the reduced rate of apoptosis in embryos cultured with beta-ME confirms its role in protecting vitrified blastocyst against stress. In summary, vitrification alters the expression of the adhesion related genes in vitrified blastocyst, which may explain, at least in part, the reason for the low pregnancy rate following transfer of such embryos into recipient animal, and the supplementation of IVC media with beta-ME significantly improved the quality of vitrified blastocyst evidenced by the modulation of the expression of blastocyst important genes, beta-catenin, E-cadherin and Oct-4, and the ability to protect vitrified blastocyst against apoptosis. (C) 2018 Elsevier Inc. All rights reserved.
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