Kovo Michal, Kandli-Cohen Miri, Ben-Haim Miri, Galiani Dalia, Carr Dan W, Dekel Nava
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot, 76100 Israel.
Reproduction. 2006 Jul;132(1):33-43. doi: 10.1530/rep.1.00824.
Reinitiation of meiosis in meiotically competent, fully grown mammalian oocytes is governed by a fall in intraoocyte cAMP concentrations and the subsequent inactivation of protein kinase A (PKA). A similar reduction in intraoocyte cAMP concentrations in growing, meiotically incompetent rat oocytes not leading to resumption of meiosis, questions the involvement of PKA in the regulation of meiosis at this early stage of oocyte development. We examined the possibility of whether PKA activity maintains growing oocytes in meiotic arrest and further explored the mode of activation of PKA under conditions of relatively low cAMP concentrations. Our experiment demonstrated that inactivation of PKA stimulates growing rat oocytes to resume meiosis, and elevates the activity of their maturation-promoting factor (MPF). We also found that the expressions of type I and type II regulatory subunits (RI and RII) of PKA are higher in growing and fully grown oocytes, respectively. In addition, we revealed that the common 1:1 ratio between the regulatory (R) and catalytic (C) subunits of PKA is apparently not abrogated and, in accordance PKA activity in growing oocyte-cell extract is fully dependent on cAMP. Finally, we identified in growing oocytes, the A kinase anchoring protein (AKAP) 140, which was previously depicted in fully grown oocytes. We conclude that an active PKA prevents growing oocytes from resuming meiosis. Our findings further suggest that relatively high abundance of the PKAI isoform and/or its subcellular compartmentalization, through interaction with AKAP140, could possibly account for the high basal PKA activity at relatively low intraoocyte cAMP concentrations.
在具有减数分裂能力、完全成熟的哺乳动物卵母细胞中,减数分裂的重新启动受卵母细胞内cAMP浓度下降以及随后蛋白激酶A(PKA)失活的调控。在生长中的、减数分裂无能力的大鼠卵母细胞中,卵母细胞内cAMP浓度有类似降低,但未导致减数分裂恢复,这对PKA在卵母细胞发育这一早期阶段减数分裂调控中的作用提出了质疑。我们研究了PKA活性是否维持生长中卵母细胞减数分裂停滞的可能性,并进一步探讨了在相对低cAMP浓度条件下PKA的激活模式。我们的实验表明,PKA失活会刺激生长中的大鼠卵母细胞恢复减数分裂,并提高其成熟促进因子(MPF)的活性。我们还发现,PKA的I型和II型调节亚基(RI和RII)的表达分别在生长中和完全成熟的卵母细胞中更高。此外,我们发现PKA调节亚基(R)和催化亚基(C)之间常见的1:1比例显然未被废除;相应地生长中卵母细胞提取物中的PKA活性完全依赖于cAMP。最后我们在生长中的卵母细胞中鉴定出了A激酶锚定蛋白(AKAP)140,该蛋白先前在完全成熟的卵母细胞中有所描述。我们得出结论,活跃的PKA可防止生长中的卵母细胞恢复减数分裂。我们的研究结果进一步表明,PKA I亚型相对较高的丰度和/或其亚细胞区室化,通过与AKAP140相互作用,可能是卵母细胞内cAMP浓度相对较低时PKA基础活性较高的原因。