Cytoplasmic membrane activities during first cleavage of zona-free porcine embryos: description and consequences

Abstract

Already at first embryo cleavage subsequent blastocyst formation can be predicted based on morphology but the finer morphological details can be difficult to determine due to the presence of the zona pellucida (ZP). Therefore, we monitored zona-free porcine parthenogenetically activated (PA) embryos in a time-lapse system to: (1) describe and characterise the morphological activity of the cytoplasmic membrane and the distribution to the two nuclei during first cleavage and (2) determine the relationship between specific morphological activities and subsequent embryonic development. After ZP removal the membrane surface activities were clearly visible, so all cleaved embryos could be divided into two groups depending on the surface activity during first cleavage: regular morphology (MN) or irregular morphology with ‘bumps’ (MB). The two nuclei were more unequal in MB embryos in both nucleus size and DNA quantity. After first cleavage, MB embryos could be further divided into three types of irregularities (MB1, MB2, MB3) based on their subsequent behaviour. Clear differences in developmental patterns were found between MN and MB embryos, such as delayed first cleavage, compromised blastocyst formation and total cell number. The predictive value of these new types of morphological events was comparable to the more traditionally used time of first cleavage. In conclusion, zona-free embryos allow visualisation of finer morphological details that can provide an early prediction of embryo developmental potential, but further studies are needed on other type of embryos.