Bellé R, Mulner-Lorillon O, Marot J, Ozon R
Cell Differ. 1986 Dec;19(4):253-61. doi: 10.1016/0045-6039(86)90102-8.
Progesterone induces in vitro the meiotic cell division of Xenopus full-grown oocytes. Microinjection into oocyte of a solution containing Mg2+ (20 mM) facilitates by one order of magnitude the dose of progesterone which induces 50% of germinal vesicle breakdown. Microinjected in the absence of hormone, Mg2+ and also Mn2+ can induce maturation with efficiencies of, respectively, 24% (SEM = 8; n = 13) and 70% (SEM = 6; n = 23). The dose-response curves of cation-induction of maturation show an optimum of 20 mM for Mg2+ and 15 mM for Mn2+ (pipet concentration); higher doses were less active. Cation-induction of maturation is inhibited when oocytes are preincubated with cholera toxin (500 ng/ml); nevertheless, it cannot be interpreted at the level of cAMP, since both Mg2+ and Mn2+ microinjections provoke an increase in the oocyte cAMP content. Mg2+ induction of maturation is more efficient when oocytes are incubated in trimethylamine at pH 8.2, which is known to increase intracellular pH suggesting an action at the level of alkali pH-sensitive enzymes. Altogether, our results indicate a positive role for Mg2+ ions in the induction of oocyte maturation and raise an attractive hypothesis about the respective roles of cAMP and Mg2+ changes involved in the mechanism of progesterone action. Our results also show that co-injection of 2-glycerophosphate and Mg2+ ions, which are both commonly used in the preparation of the MPF mitotic factors from dividing cells, induces oocyte maturation more efficiently than Mg2+ alone and drastically shortens the kinetics of germinal vesicle breakdown to 1 h 30 min to 2 h 30 min.
孕酮可在体外诱导非洲爪蟾完全成熟的卵母细胞进行减数分裂。向卵母细胞显微注射含Mg2+(20 mM)的溶液,可使诱导50%生发泡破裂的孕酮剂量提高一个数量级。在无激素的情况下显微注射,Mg2+以及Mn2+均可诱导成熟,其效率分别为24%(标准误=8;n=13)和70%(标准误=6;n=23)。阳离子诱导成熟的剂量-反应曲线显示,Mg2+的最佳浓度为20 mM,Mn2+为15 mM(移液管浓度);更高剂量的活性较低。当卵母细胞与霍乱毒素(500 ng/ml)预孵育时,阳离子诱导的成熟受到抑制;然而,这无法在cAMP水平进行解释,因为Mg2+和Mn2+显微注射均会引起卵母细胞cAMP含量增加。当卵母细胞在pH 8.2的三甲胺中孵育时,Mg2+诱导成熟的效率更高,已知三甲胺可提高细胞内pH,提示其作用于碱pH敏感酶水平。总之,我们的结果表明Mg2+离子在诱导卵母细胞成熟中具有积极作用,并提出了一个关于cAMP和Mg2+变化在孕酮作用机制中各自作用的有趣假设。我们的结果还表明,共同注射常用于从分裂细胞制备MPF有丝分裂因子的2-甘油磷酸和Mg2+离子,比单独注射Mg2+更有效地诱导卵母细胞成熟,并将生发泡破裂的动力学急剧缩短至1小时30分钟至2小时30分钟。