Funahashi H, Cantley T C, Day B N
Department of Animal Sciences, University of Missouri-Columbia, 65211, USA.
Biol Reprod. 1997 Jul;57(1):49-53. doi: 10.1095/biolreprod57.1.49.
The effect of stage of maturation of the germinal vesicle of porcine oocytes at the time of in vitro maturation on subsequent developmental competence was examined. A large variation exists in the germinal vesicle morphology of oocytes at the time of collection of cumulus-oocyte complexes (COCs) and after culture in the absence of dibutyryl cAMP (dbcAMP) for 20 h. However, the morphology of the germinal vesicle was synchronized to a specific stage after culture in the presence of 1 mM dbcAMP for 20 h. There was no difference in germinal vesicle breakdown rate (total mean, 75.0 +/- 5.4%) or in maturation rate (total mean, 82.1 +/- 2.1 %) at 28 and 44 h of culture, respectively. However, differences in meiotic progress of oocytes were observed (p < 0.05) at 36 h of culture when COCs were exposed to dbcAMP for the first 20 h of maturation, as compared to controls. The incidence of embryos that developed to the blastocyst stage after in vitro fertilization was higher (p < 0.05) when COCs were exposed to dbcAMP (21.5 +/- 2.5%) as compared to controls (9.2 +/- 1.6%). After transfer of experimental embryos to four recipient gilts, the three pregnant recipients delivered 19 live piglets. These results indicate that exposure of COCs to dbcAMP for the first 20 h of culture for maturation increases the homogeneity of oocyte nuclear maturation and improves the efficiency of in vitro production of swine embryos.
研究了猪卵母细胞体外成熟时生发泡成熟阶段对后续发育能力的影响。在收集卵丘 - 卵母细胞复合体(COCs)时以及在无二丁酰环磷腺苷(dbcAMP)的情况下培养20小时后,卵母细胞的生发泡形态存在很大差异。然而,在含有1 mM dbcAMP的情况下培养20小时后,生发泡的形态同步到了特定阶段。在培养28小时和44小时时,生发泡破裂率(总平均值,75.0 +/- 5.4%)和成熟率(总平均值,82.1 +/- 2.1 %)分别没有差异。然而,与对照组相比,当COCs在成熟的前20小时暴露于dbcAMP时,在培养36小时时观察到卵母细胞减数分裂进程存在差异(p < 0.05)。与对照组(9.2 +/- 1.6%)相比,当COCs暴露于dbcAMP(21.5 +/- 2.5%)时,体外受精后发育到囊胚阶段的胚胎发生率更高(p < 0.05)。将实验胚胎移植到4头受体母猪后,3头怀孕的受体母猪产下了19头活仔猪。这些结果表明,在培养成熟的前20小时将COCs暴露于dbcAMP可增加卵母细胞核成熟的同质性,并提高猪胚胎体外生产的效率。