Stock A E, Woodruff T K, Smith L C
Centre de recherche en reproduction animale, Faculté de Médecine Vétérinaire, St. Hyacinthe, Québec, Canada.
Biol Reprod. 1997 Jun;56(6):1559-64. doi: 10.1095/biolreprod56.6.1559.
The aim of this study was to investigate whether inhibin A and/or activin A play a role in the acquisition of oocyte competence during the final stages of oogenesis. The particular goal was to establish whether inhibin A and activin A exert development-enhancing effects during in vitro maturation in serum-free media and whether such effects are related to changes in the kinetics of meiotic resumption and/or fertilization rates. Cumulus-oocyte-complexes (COCs) were matured in two control media (Medium 199 [M199] with hormones and serum, hormone-serum control; M199 + 0.6% BSA, BSA-control) and nine treatment media (M199 with 0.6% BSA containing 100, 10, and 1 ng/ml of recombinant human inhibin A, recombinant human activin A, and the combination of the two). Oocytes were fertilized and cultured using standard procedures. Cleavage was assessed at 54 h and blastocyst development at 8 days after in vitro fertilization. Kinetics of oocyte maturation and the fertilization rates were evaluated after fixing and staining (Hoechst 33342) of oocytes at 8, 16, and 22 h after onset of in vitro maturation or of presumptive zygotes at 12 h after in vitro fertilization, respectively. Although there was no effect on cleavage rates, inhibin A and activin A significantly enhanced postcleavage development at concentrations of 10 ng/ml (57.7 +/- 7.5% and 56.6 +/- 11.7%, respectively) and 100 ng/ml (50.6 +/- 18.6% and 56.4 +/- 4.0%, respectively) compared to that in the BSA-control group (24.6 +/- 3.2%). Whereas inhibin A- and activin A-treated oocytes showed development-enhancing effects similar to those in the hormone-serum controls, these groups differed with regard to the kinetics of meiotic resumption. Likewise, the enhanced development of the hormone-serum control and the inhibin A/activin A-treated oocytes was not related to increased fertilization rates relative to the BSA-control. These results suggest that inhibin A and activin A may play important roles during the final stages of oogenesis and that recombinant inhibins and activins are useful compounds for the development of a serum-free culture system for in vitro maturation of oocytes from cattle and possibly other mammalian species.
本研究的目的是调查抑制素A和/或激活素A在卵子发生最后阶段卵母细胞能力获得过程中是否发挥作用。具体目标是确定抑制素A和激活素A在无血清培养基中体外成熟过程中是否发挥促进发育的作用,以及这种作用是否与减数分裂恢复动力学和/或受精率的变化有关。卵丘-卵母细胞复合体(COCs)在两种对照培养基(含激素和血清的199培养基[M199],激素-血清对照;M199 + 0.6%牛血清白蛋白,牛血清白蛋白对照)和九种处理培养基(含0.6%牛血清白蛋白的M199,分别含有100、10和1 ng/ml重组人抑制素A、重组人激活素A以及两者的组合)中成熟。使用标准程序对卵母细胞进行受精和培养。在体外受精后54小时评估卵裂情况,在8天时评估囊胚发育情况。分别在体外成熟开始后8、16和22小时或体外受精后12小时对卵母细胞或推测的受精卵进行固定和染色(Hoechst 33342)后,评估卵母细胞成熟动力学和受精率。虽然对卵裂率没有影响,但与牛血清白蛋白对照组(24.6 +/- 3.2%)相比,抑制素A和激活素A在浓度为10 ng/ml(分别为57.7 +/- 7.5%和56.6 +/- 11.7%)和100 ng/ml(分别为50.6 +/- 18.6%和56.4 +/- 4.0%)时显著增强了卵裂后的发育。虽然经抑制素A和激活素A处理的卵母细胞显示出与激素-血清对照组相似的促进发育作用,但这些组在减数分裂恢复动力学方面存在差异。同样,激素-血清对照组和经抑制素A/激活素A处理的卵母细胞发育增强与相对于牛血清白蛋白对照组受精率增加无关。这些结果表明,抑制素A和激活素A可能在卵子发生的最后阶段发挥重要作用,并且重组抑制素和激活素是开发用于牛及可能其他哺乳动物物种卵母细胞体外成熟的无血清培养系统的有用化合物。