Plasma membrane and cytocortex alterations in frozen/thawed mouse embryos deprived of the zona pellucida

We analyzed the morphological and ultrastructural abnormalities present in two-cell mouse embryos frozen/thawed without the zona pellucida (ZP). Transmission electron microscopy revealed that these embryos had plasma membrane abnormalities that were not observed in the embryos frozen/thawed with an intact ZP. The most frequent anomalies were a decreased number of microvilli with a nonhomogeneous distribution and showing an abnormal morphology and the presence of an increased number of vesicles in the periphery of the cell. The distribution of actin filaments, studied by fluorescence microscopy revealed alterations in both embryos, frozen/thawed with ZP and embryos frozen/thawed without ZP: an increased staining in some regions of the peripheral actin band, discontinuities or gaps in the band, or the presence of a second actin band connected to the peripheral one.