Nemcova Lucie, Machatkova Marie, Hanzalova Katerina, Horakova Jindra, Kanka Jiri
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Rumburská 89, 277 21 Libechov, Czech Republic.
Theriogenology. 2006 Apr 15;65(7):1254-64. doi: 10.1016/j.theriogenology.2005.08.006. Epub 2005 Sep 19.
The aim of this study was to compare the expression of selected genes in bovine embryos developed from oocytes with different developmental competence. Four oocyte populations were collected, separately either from small (2-5 mm) or medium (6-10 mm) follicles, in the growth/stagnation (G/S) or dominance/regression (D/R) stage of the first follicular wave. They were matured, fertilized and cultured to D7 or D8 blastocysts by a standard protocol. Poly (A)+ mRNA was extracted from pooled blastocysts and the expression of bax-alpha (Bax), connexin 43 (Cx 43) and connexin 31 (Cx 31) was estimated using real-time RT-PCR. The cleavage rates were significantly higher in oocytes collected from both medium and small follicles, (p < or = 0.05 and p < or = 0.01, respectively) in the G/S than in the D/R stage. There were no significant differences in the D7 blastocyst rates between oocytes from both medium and small follicles in the G/S or D/R stage. But the D8 blastocyst rate was significantly higher in oocytes from small follicles in the G/S stage compared with those in the D/R stage. The relative abundance of Bax and Cx 31 made no significant difference in both D7 and D8 blastocysts developed from oocytes collected from medium or small follicles in the G/S or D/R stages. But the relative abundance of the Cx 43 transcript was significantly higher in D8 blastocysts developed from oocytes collected from both medium and small follicles in the G/S stage compared with those in the D/R stage. We conclude that the relative abundance of Cx 43 can be used as a marker of developmental potential for embryos derived from oocytes with different developmental competence because the level of Cx 43 transcript was greater in embryos derived from oocytes with greater developmental competence compared with those derived from oocytes with lesser developmental competence.
本研究旨在比较不同发育能力的卵母细胞发育而来的牛胚胎中特定基因的表达情况。收集了四个卵母细胞群体,分别来自小卵泡(2 - 5毫米)或中等卵泡(6 - 10毫米),处于第一个卵泡波的生长/停滞(G/S)或优势/退化(D/R)阶段。按照标准方案将它们进行成熟、受精并培养至第7天或第8天的囊胚。从合并的囊胚中提取多聚腺苷酸加尾(Poly (A)+)mRNA,并使用实时逆转录聚合酶链反应(real-time RT-PCR)评估促凋亡蛋白bax-alpha(Bax)、连接蛋白43(Cx 43)和连接蛋白31(Cx 31)的表达。在G/S阶段,从中等卵泡和小卵泡收集的卵母细胞的分裂率显著更高(分别为p≤0.05和p≤0.01),高于D/R阶段。在G/S或D/R阶段,中等卵泡和小卵泡的卵母细胞在第7天的囊胚率没有显著差异。但与D/R阶段相比,G/S阶段小卵泡的卵母细胞在第8天的囊胚率显著更高。在G/S或D/R阶段,从中等卵泡或小卵泡收集的卵母细胞发育而来的第7天和第8天囊胚中,Bax和Cx 31的相对丰度没有显著差异。但与D/R阶段相比,G/S阶段从中等卵泡和小卵泡收集的卵母细胞发育而来的第8天囊胚中,连接蛋白43转录本的相对丰度显著更高。我们得出结论,Cx 43的相对丰度可作为不同发育能力的卵母细胞来源胚胎发育潜力的标志物,因为与发育能力较低的卵母细胞来源的胚胎相比,发育能力较高的卵母细胞来源的胚胎中Cx 43转录本水平更高。