Department of Biological Sciences, Graduate School of Science, Osaka University, Toyonaka, Osaka, 560-0043, Japan.
Department of Biological Sciences, Graduate School of Science, Osaka University, Toyonaka, Osaka, 560-0043, Japan.
Dev Biol. 2020 Apr 15;460(2):155-163. doi: 10.1016/j.ydbio.2019.12.005. Epub 2019 Dec 16.
Unfertilized eggs of most animals are arrested at a certain point in the meiotic cell cycles. Reinitiation of meiosis and the start of embryogenesis are triggered by fertilization. This arrest is essential for preventing parthenogenetic activation and for promoting proper initiation of development by fertilization. In the larvacean Oikopleura dioica, which is a simple model organism for studies of chordate development, the unfertilized egg is arrested at metaphase of meiosis I. We show here that protein phosphatase 2A (PP2A) is essential for maintenance of meiotic arrest after spawning of oocytes. Knockdown (KD) of the maternal PP2A catalytic subunit, which was found in functional screening of maternal factors, caused unfertilized eggs to spontaneously release polar bodies after spawning, and then start pseudo-cleavages without fertilization, namely, parthenogenesis. Parthenogenetic embryos failed to undergo proper mitosis and cytokinesis because of lack of a centrosome, which is to be brought into the egg by a sperm. Activation of the KD oocytes was triggered by possible rise of ambient and intracellular pH upon their release from the gonad into seawater at spawning. Live recording of intracellular calcium level of the KD oocytes indicated that the pH rise caused an aberrant Ca burst, which mimicked the Ca burst that occurs at fertilization. Then, the aberrant Ca burst triggered meiosis resumption through Calcium/calmodulin-dependent protein kinase (CaMK II). Therefore, PP2A is essential for maintenance of meiotic arrest and prevention of parthenogenesis by suppressing the aberrant Ca burst at spawning.
大多数动物的未受精卵在减数分裂细胞周期的某个特定点停滞。受精触发减数分裂的重新启动和胚胎发生的开始。这种停滞对于防止孤雌激活和促进受精后适当的发育启动是必不可少的。在长尾幼虫 Oikopleura dioica 中,它是研究脊索动物发育的简单模式生物,未受精卵在减数分裂 I 的中期停滞。我们在这里表明,蛋白磷酸酶 2A(PP2A)对于卵母细胞产卵后维持减数分裂停滞是必不可少的。在母体因子的功能筛选中发现的母体 PP2A 催化亚基的敲低(KD)导致未受精卵在产卵后自发释放极体,然后在没有受精的情况下开始假性分裂,即孤雌生殖。由于缺乏中心体,孤雌生殖胚胎无法进行适当的有丝分裂和胞质分裂,中心体是由精子带入卵子的。KD 卵母细胞的激活是由它们从性腺释放到海水中时周围和细胞内 pH 值的可能升高引发的。KD 卵母细胞的细胞内钙离子水平的实时记录表明,pH 值的升高导致异常的 Ca 爆发,这模拟了受精时发生的 Ca 爆发。然后,异常的 Ca 爆发通过钙/钙调蛋白依赖性蛋白激酶(CaMK II)触发减数分裂恢复。因此,PP2A 通过抑制产卵时的异常 Ca 爆发对于维持减数分裂停滞和防止孤雌生殖是必不可少的。