Palasz A T, Rodriguez-Martinez H, Beltran-Breña P, Perez-Garnelo S, Martinez M F, Gutierrez-Adan A, De la Fuente J
Departamento de Reproducción Animal y Conservación de Recursos Zoogenéticos, INIA, Madrid, Spain.
Mol Reprod Dev. 2006 Dec;73(12):1503-11. doi: 10.1002/mrd.20516.
Effects of hyaluronan (HA), BSA, and FCS on in vitro development, ultrastructure, and mRNA transcription of four developmentally important genes: apoptosis (Bax), oxidative stress (SOX), growth factor (IGF-II), and cell-to-cell adhesion (Ecad) were examined. Two biological origin HA, Hylartil and Hyonate and one produced by fermentation (f-HA) MAP-5 were tested. Embryos were cultured in SOF medium with 0.4% BSA or with 0.4% BSA and 10% FCS. HA was added 96 hr post insemination (pi) to half of the embryos from each culture group. Embryo development was not affected by either HA preparation, however, hatching rates were higher in Hyalartil and MAP-5 than in control and Hyonate (P < 0.05). There was no effect of HA on number of blastocysts developed in SOF + BSA. However, more blastocysts developed in SOF + BSA + f-HA than in SOF + BSA + FCS or with BSA + FCS + f-HA. HA added to SOF + BSA, increased level of expression of epidermal growth factor (EGF)-II and decreased the levels of expression of BAX, SOX, and Ecad (P < 0.05). Presence of FCS increased the levels of SOX and decreased the level of IGF-II (P < 0.05) and the addition of f-HA to SOF containing FCS showed no effect on the level of transcription of any analyzed genes. The fine structure of embryos cultured with f-HA irrespective of protein sources used was clearly improved. In summary, f-HA added 96 hr pi to SOF supplemented with BSA but not FCS improved development, molecular composition and fine structure of bovine embryos.
研究了透明质酸(HA)、牛血清白蛋白(BSA)和胎牛血清(FCS)对四个发育重要基因:凋亡基因(Bax)、氧化应激基因(SOX)、生长因子基因(IGF-II)和细胞间黏附基因(Ecad)的体外发育、超微结构及mRNA转录的影响。测试了两种生物来源的HA,即Hylartil和Hyonate,以及一种发酵产生的(f-HA)MAP-5。胚胎在含有0.4% BSA或含有0.4% BSA和10% FCS的SOF培养基中培养。授精后96小时(pi),将HA添加到每个培养组一半的胚胎中。两种HA制剂均未影响胚胎发育,然而,Hylartil和MAP-5组的孵化率高于对照组和Hyonate组(P < 0.05)。HA对在SOF + BSA中发育的囊胚数量没有影响。然而,在SOF + BSA + f-HA中发育的囊胚比在SOF + BSA + FCS或BSA + FCS + f-HA中更多。添加到SOF + BSA中的HA增加了表皮生长因子(EGF)-II的表达水平,并降低了BAX、SOX和Ecad的表达水平(P < 0.05)。FCS的存在增加了SOX的水平并降低了IGF-II的水平(P < 0.05),并且向含有FCS的SOF中添加f-HA对任何分析基因的转录水平均无影响。无论使用何种蛋白质来源,用f-HA培养的胚胎的精细结构都明显改善。总之,授精后96小时向补充有BSA而非FCS的SOF中添加f-HA可改善牛胚胎的发育、分子组成和精细结构。