Park J E, Oh H J, Hong S G, Jang G, Kim M K, Lee B C
Department of Theriogenology and Biotechnology, SeoulNational University, Seoul, Korea.
Reprod Domest Anim. 2010 Aug;45(4):585-93. doi: 10.1111/j.1439-0531.2008.01306.x. Epub 2008 Dec 15.
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.
本研究的目的是评估激活素A对体外受精(IVF)牛胚胎发育能力的影响,这些胚胎来自两步限定培养系统(C1/C2培养基),处于胚胎发育的早期或后期。为了评估激活素A对转录水平的影响,我们分析了与体外受精胚胎中囊胚孵化和着床以及激活素信号通路相关的基因。卵丘-卵母细胞复合体成熟22小时后进行体外受精。假定的合子在早期(0-120小时,C1)或后期(120-192小时,C2)阶段在有或没有激活素A的情况下培养。虽然在C1培养基中用激活素A培养的胚胎的发育能力与其相应对照组没有显著差异,但在添加100 ng/ml激活素A的C2培养基中,发育到囊胚的比例(22.4%对34.7%;p<0.05)和囊胚孵化率(9.3%对22.4%;p<0.05)显著高于对照组。为了评估激活素A对转录的影响,分析了与囊胚孵化和着床相关的基因(Na/K-ATP酶、E-钙黏蛋白和葡萄糖转运蛋白1)以及激活素信号通路(激活素受体II、激活素受体IIB和Smad2)的相对表达水平。与对照培养基相比,添加激活素A的培养基中Na/K-ATP酶、E-钙黏蛋白、葡萄糖转运蛋白1、激活素受体II和激活素受体IIB的基因表达增加。总之,本研究表明,在体外牛胚胎发育的后期,激活素A可以通过提高孵化率和影响限定培养基中与孵化和着床相关基因的表达水平来增强胚胎的体外发育。