Structural and Functional Events on the Porcine Zona Pellucida During Maturation, Fertilization and Embryonic Development: a Scanning Electron Microscopy Analysis

Abstract

Porcine oocytes and pre-implantation embryos from the same, as well as from different animals, have an extremely heterogeneous morphology of the zona pellucida (ZP) surface, as shown by scanning electron microscopy. For years, it has been believed that this heterogeneous morphology plays an important role in the sperm-oocyte interaction. The aim of this study was to analyse the zona morphology and sperm-binding patterns on the porcine ZP. Oocytes were divided into four categories: immature, matured in vivo, or matured in vitro over a time period of 24 or 48 h. The zona morphology of early embryos grown in vivo or in vitro was also investigated. Four different types of zona morphology were detectable. They ranged from a porous, net-like structure to a nearly smooth and compact surface. No correlation could be established between the different kinds of maturation in terms of these zona types. All oocytes exhibited extremely heterogeneous zona morphologies, with no clear trend. During subsequent in vivo embryo development, the zona surface changes from a porous structure to one with a compact surface, while the morphology of in vitro embryos remained compact at all stages of development. The analysis of the number and distribution patterns of spermatozoa trapped in the ZP revealed extremely variable patterns, regardless of the zona morphology. Differences were only present if sorted or unsorted spermatozoa were used for insemination. Regardless of the number of inseminated spermatozoa after sorting, only a few (1-2) could be detected on the ZP. Whether oocytes were matured in vivo or in vitro was not a relevant factor. Unsorted spermatozoa bound in higher numbers than sorted ones. The number was directly dependent on the number of spermatozoa used for insemination.