Autonomous cortical activity in mouse eggs controlled by a cytoplasmic clock

Abstract

Mouse eggs of Swiss albino origin, both parthenogenetic and fertilized, were bisected into nucleate (NHs) and anucleate halves (AHs) and observed in vitro (semicontinuous observations) for up to 40 h for possible manifestations of cortical activity. Three experimental groups were studied: (1) Non-fertilized eggs activated 17 h after administration of hCG with a heat-shock and bisected 4 h later. (2) Non-fertilized eggs first bisected, and the resulting sister halves activated 17 h after administration of hCG with ethyl alcohol. (3) In vivo fertilized eggs bisected 27 h after administration of hCG into an AH and a binucleate half. Parthenogenetic eggs (intact, zona-free, and incompletely bisected), and fertilized eggs collected 17, 20, and 27 h after administration of hCG were also studied. In the middle of the first cell cycle the cell surface in all types of cells studied changed from smooth to slightly undulate. In nucleate cells the surface deformations lasted for several hours and disappeared shortly before the first mitosis. In contrast, in AHs the indentations of the cell surface deepened, and developed into manifold furrows, thus leading to fragmentation. However, in 20% of AHs fragmentation was partially or completely reversed. The incidence and the intensity of fragmentation were lower, and its reversibility was more common in AHs carrying the 2nd polar body. We suggest that the interphase nucleus, i.e. the pronucleus in whole eggs and NHs, and the 2nd polar body nucleus (if 2nd polary body is attached to an AH) exerts a moderating effect on cortical activity. However, the initiation of cortical activity is nucleus-independent, as shown by the behaviour of AHs separated before activation. We believe that the observed phenomena reflect autonomous cortical activity which is regulated by a cytoplasmic clock.