Effects of Activin A on theIn VitroDevelopment and mRNA Expression of Bovine Embryos Cultured in Chemically-Defined Two-Step Culture Medium

Abstract

The purpose of the present study was to evaluate the effects of activin A on the developmental competence of in vitro fertilized (IVF) bovine embryos derived from a two-step defined culture system (C1/C2 medium) during the early or later stages of embryo development. To evaluate the effects of activin A on transcriptional levels, we analysed genes related to blastocyst hatching and implantation and to activin signalling pathway in IVF embryos. Cumulus-oocyte complexes were matured for 22 h and fertilized in vitro. Presumptive zygotes were cultured in the presence or absence of activin A during early (0-120 h, C1) or later (120-192 h, C2) stages. Although the developmental competence of embryos cultured with activin A in C1 medium was not significantly different from their corresponding controls, development to blastocysts (22.4% vs 34.7%; p < 0.05) and the blastocyst hatching rate (9.3% vs 22.4%; p < 0.05) in C2 medium supplemented with 100 ng/ml activin A were significantly higher than in the control group. To evaluate the effect of activin A on transcription, the relative expression levels of genes related to blastocyst hatching and implantation (Na/K-ATPase, E-cad and Glut-1) as well as activin signalling pathway (ActRII, ActRIIB and Smad2) were analysed. Compared to control medium, gene expression of Na/K-ATPase, E-cad, Glut-1, ActRII and ActRIIB was increased in medium supplemented with activin A. In conclusion, this study suggests that activin A, during the later stage of in vitro bovine embryo development, can enhance in vitro development of embryos by increasing hatching rates and affecting expression levels of genes related to hatching and implantation in defined culture medium.