EFFECT OF ELECTROPORATION ON THE EMBRYONIC DEVELOPMENT CAPACITY OF PORCINE OOCYTES AND ZYGOTES
Supporting Agencies
- This work has been funded by the PID2022-138666OB-100 project of the Ministry of Science, Innovation and Universities.
Abstract
The aim of this study was to evaluate the impact of electroporation on in vitro matured (MII) oocytes and in vitro produced zygotes (ZG) at 2- and 5-hours post in vitro fertilization (IVF). Oocytes collected from prepubertal gilts were matured in vitro. A total of 3485 MII oocytes were used in this study. Electroporation was performed in Opti-MEM 1 medium without gene editors using either 25 or 30 V. The first group of MII was electroporated at the end of the maturation period, half of them were subjected to IVF, while the other half was incubated in IVF medium without sperm to assess parthenogenesis. The remaining oocytes were electroporated either 2 h (ZG-2) or 5 h (ZG-5) after the onset of sperm co-incubation. Controls were non-electroporated oocytes subjected to IVF. Presumptive zygotes from all groups were cultured for 7 days. On day 2 of in vitro culture the embryonic cleavage rate and the rate of degenerated oocytes were assessed. At day 7 of IVC the blastocyst formation rate and the efficiency were calculated. The total cell number and apoptosis index of day 7 blastocysts were assessed. The electroporation produced an increased (p<0.05) percentage of degenerated oocytes. Electroporation induced the parthenogenetic activation of MII resulting in similar embryonic development to that of their fertilized counterparts. The electroporation of ZG-2-30 showed a decrease (p<0.05) in the blastocyst formation rate, while the other groups of zygotes kept comparable results to controls (40.90 ± 2.80%). In conclusion, electroporation of zygotes at 5 h (either with 25 or 30 V) and at 2 h (using 25 V) after IVF supports similar embryo development to that of controls. In contrast, electroporation of MII oocytes impairs developmental potential and induces parthenogenetic activation, resulting in developmental outcomes like those of fertilized oocytes.
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