Scanning electron microscopy of in vitro fertilization in cattle

Cumulus-oocyte complexes were collected from cows at an abattoir by aspiration from small (1-6 mm) follicles. The complexes were matured in vitro for 28 h. Subsequently, the cumulus cells and the zona pellucida were removed by enzyme treatment in a proportion of the complexes (zona-free ova). Both cumulus-enclosed and zona-free ova were inseminated in vitro and processed for scanning electron microscopy after different periods of culture. In the cumulus-enclosed ova the number of spermatozoa attached to and penetrating into the cumulus investment increased with increasing culture time. Practically all spermatozoa displayed intact acrosomes. In the zona-free ova clusters of spermatozoa attached to the ovum surface, and at 5 h a proportion of the spermatozoa had undergone the acrosome reaction, and their internalization into the ooplasma was initiated. The acrosome reaction was characterized by an increasing fenestration of the membrane coverings of the acrosomal region of the sperm head. During the sperm head internalization, where the ovum microvilli appeared to contact especially the equatorial segment and the postacrosomal region, the sperm head gradually disappeared from the ovum surface, and the microvilli at the site of internalization became more bulbous. Simultaneous abstriction of the second polar body was seen in some ova.