Bovine Oocyte Quality in Relation to Ultrastructural Characteristics of Zona Pellucida, Polyspermic Penetration and Developmental Competence

Bovine Oocyte Quality in Relation to Ultrastructural Characteristics of Zona Pellucida, Polyspermic Penetration and Developmental Competence.

P Santos niversity of the Azores, Department of Agrarian Sciences, CITA-A, Animal Reproduction, Angra do Heroismo, Portugal,
A Chaveiro niversity of the Azores, Department of Agrarian Sciences, CITA-A, Animal Reproduction, Angra do Heroismo, Portugal,
N Simões niversity of the Azores, Department of Biology, Ponta Delgada, Açores, Portugal and
F Moreira da Silva niversity of the Azores, Department of Agrarian Sciences, CITA-A, Animal Reproduction, Angra do Heroismo, Portugal
University of the Azores, Department of Agrarian Sciences, CITA-A, Animal Reproduction, Angra do Heroismo, Portugal and University of the Azores, Department of Biology, Ponta Delgada, Açores, Portugal.

In the present study, the effect of bovine oocyte quality related to ultrastructural characteristics of zona pellucida (ZP), polyspermic penetration and embryo developmental competence was evaluated. Cumulus–oocyte complexes were punctioned from 453 ovaries, classified as 1, 2, 3 and 4 according to their morphological aspect, matured for 24 h and then divided into two groups. In group A, oocytes were fixed in 2.5% glutaraldehyde and 0.1 m sodium cacodylate and examined under a scanning electron microscope. Photomicrographs were taken and ZP’s pores were evaluated in squares of 6.4-μm width. In group B, oocytes were fertilized in vitro. After 48 h, non-cleaved oocytes were fixed for polyspermy evaluation. On days 7, 9 and 10, embryos were classified as developed (blastocysts and hatched blastocysts). Results showed that quality 1 oocytes revealed a ZP pore diameter of 0.50 ± 0.07 μm, which was smaller than the observed on oocytes of quality 2 (0.83 ± 0.10 μm), quality 3 (1.02 ± 0.22 μm) and quality 4 (1.38 ± 0.59 μm) (p ≤ 0.05). For In Vitro Fertilization (IVF), results showed that embryos originating from oocytes classed as 3 and 4 had lower cleavage rate (68.4% and 43.8%) than those belonging to class 1 and 2 (79.5% and 69.3%) (p ≤ 0.05). None oocyte classified as 3 and 4 developed to hatch blastocysts, while for oocytes belonging to quality 1 and 2, these values were, respectively, 15.2% and 12.5%. Concerning polyspermy, oocytes class 1 and 2 had lower polyspermic penetration than those belonging to class 3 and 4 (respectively 4.1%, 4.5%, 11.1% and 9.8%, for class 1, 2, 3 and 4). In conclusion, the present study demonstrated that oocytes with low qualities result in lower developmental competence and with high percentage of polyspermy after IVF, which can be the result of the ZP structure such as the number and the pore’s diameter.

(In Reproduction in domestic animals)