Genética y Reproducción Animal, Centro de Biotecnología Animal, SERIDA, Camino de Rioseco 1225, La Olla – Deva, 33394 Gijón, Asturias, Spain.
Theriogenology. 2011 Feb;75(3):584-8. doi: 10.1016/j.theriogenology.2010.09.010. Epub 2010 Oct 30.
The aim of this study was to investigate the effects of activin A on development, differential cell counts and apoptosis/necrosis rates of bovine embryos produced in vitro. Presumptive zygotes were cultured up to Day 8 in synthetic oviduct fluid containing aminoacids, citrate, myo-inositol and BSA. In Experiment 1, activin (10 ng mL(-1)) was added: 1/from Day 1 to Day 3; 2/from Day 1 to Day 8; 3/from Day 3 to Day 8; or 4/absent (control). In Experiment 2, 10 ng mL(-1) activin were added either before (Day 3 to Day 5) or after (Day 5 to Day 8) the early morula stage. In Experiment 1, activin during the first 72 h of culture reduced Day 3 cleavage, 5-8 cell rates and blastocyst development, while hatching rates increased. No changes were observed within differential cell counts. In experiment 2, activin improved blastocyst development after, and had no effect before, the Day 5 morula stage. However, trophectoderm (TE) cell numbers decreased with activin both before and after the Day 5 morula stage, suggesting that activin inhibits TE differentiation. The presence of activin during the whole culture had no effect on TUNEL positive cells, but when added at shorter periods activin increased apoptotic rates. Effects of activin during in vitro bovine embryo development, depends on timing of its addition to the culture medium.
本研究旨在探讨激活素 A 对体外培养牛胚胎发育、分化细胞计数和凋亡/坏死率的影响。将假定的受精卵在含有氨基酸、柠檬酸盐、肌醇和 BSA 的合成输卵管液中培养至第 8 天。在实验 1 中,添加激活素(10ng/ml):1/从第 1 天到第 3 天;2/从第 1 天到第 8 天;3/从第 3 天到第 8 天;或 4/不存在(对照)。在实验 2 中,在早期桑椹胚阶段之前(第 3 天至第 5 天)或之后(第 5 天至第 8 天)添加 10ng/ml 的激活素。在实验 1 中,培养的前 72 小时添加激活素会降低第 3 天的卵裂率、5-8 细胞率和囊胚发育率,而孵化率增加。分化细胞计数无变化。在实验 2 中,激活素在第 5 天桑椹胚阶段之后改善了囊胚发育,但在之前没有影响。然而,激活素在第 5 天桑椹胚阶段前后都会减少滋养外胚层(TE)细胞数量,表明激活素抑制 TE 分化。激活素在整个培养过程中的存在对 TUNEL 阳性细胞没有影响,但在较短时间添加时,激活素会增加凋亡率。激活素对体外牛胚胎发育的影响取决于其添加到培养基中的时间。