Postovulatory ageing induces structural changes in the mouse zona pellucida

The structure of the zona pellucida (ZP) was analyzed in mouse oocytes collected soon after ovulation and in others retrieved 20 h after. The conventional methods for electron microscopy and the alcoholic PTA staining procedure which preferentially contrasts lysine-rich proteins were employed. The ZP of aged oocytes showed several structural changes which were particularly observed after using the PTA procedure. In 82.14% of the aged oocytes the ZP appeared clearly composed of two different regions: an inner dense and an outer of low density. The ZP showed a fibrillar banded structure with a parallel arrangement of fibrillar threads in both the outer and inner regions. The in vitro fertilization analysis showed that only 16.85% of the aged gametes attained the two cell embryo stage in comparision to 66.93% shown by the freshly ovulated eggs. The non-fertilized oocytes showed that no sperm penetration through the ZP occurred.