Cleavage characteristics of mouse embryos inseminated and cultured after zona pellucida drilling

Abstract

The effects of zona drilling on mouse embryo development in vitro were evaluated. Following insemination, sperm were immediately concentrated at the area of drilling, and in zona-drilled eggs, pronuclei appeared 30-50 min earlier than in zona-intact controls. Zona-drilled oocytes fertilized at significantly higher rates than undrilled controls and, consequently, a greater percentage of eggs inseminated after zona drilling reached the blastocyst stage. The attrition rates of zona-drilled embryos at each cleavage stage did not differ significantly from controls. Manipulated embryos exhibited unique cleavage patterns. Some embryos lost their zonae entirely, whereas others became partially extruded at early cleavage stages. These anomalies led to separation of blastomeres from the zygote proper, aggregation of embryos to form giant composite morulae and blastocysts, and occasionally to formation of miniature twin blastocysts. These characteristics of cleavage indicate that although zona drilling of a cohort of oocytes is likely to lead to an increased number of live births relative to controls, some developmental abnormalities can be encountered, and these may be associated with embryo loss, spontaneous chimerism, or possibly with conception of monozygotic twins.