Zona-free and with-zona parthenogenetic embryo production in goat (Capra hircus) - effect of activation methods, culture systems and culture media

Studies on parthenogenetic activation of oocytes are important to improve the efficiency of nuclear transfer as artificial activation of oocytes is an essential component of nuclear transfer protocol. The present study was carried out to investigate the effects of different activation methods, culture systems and culture media on in vitro development of zona-free and with-zona parthenogenetic embryos in goat. In case of zona-free parthenogenesis, there was a significant (p < 0.05) increase in cleavage rate and blastocyst yield when oocytes were activated by electrical pulse (76.29 ± 0.52% and 19.07 ± 0.39% respectively) than when Ca-ionophore was used for activation (63.45 ± 0.73% and 14.09 ± 0.65% respectively). The quality of blastocysts was evaluated by determination of cell number and by expression profile of pluripotent related gene Oct-4. No significant (p < 0.05) difference was found in quality of blastocysts produced by different activation methods. In culturing of zona-free parthenogenetic embryos, flat surface (FS) was proved to be the best system. A significant (p < 0.05) decrease in cleavage rate and blastocyst yield was found in Microdrop culture of zona-free embryos (43.67 ± 2.08% and 0.72 ± 0.72% respectively) in comparison to WOW of zona-free embryos (73.88 ± 1.70% and 15.51 ± 1.34% respectively) and FS of zona-free (75.14 ± 0.81% and 23.93 ± 2.71% respectively) as well as with-zona (72.16 ± 1.55% and 18.16 ± 0.68%) embryos. Zona-free flat culture system yielded significantly (p < 0.05) higher blastocyst rate than zona-free WOW system as well as with-zona flat culture system. The zona-free and with-zona parthenogenetic embryos were cultured in three different media - Research Vitro Cleave media from Cook® Australia (RVCL), Embryo Development Medium (EDM) and Modified Synthetic Oviductal Fluid (mSOF). In case of zona-free parthenogenesis, significant (p < 0.05) increase was found in blastocyst development rate in RVCL medium (18.61 ± 1.52%) than EDM (11.29 ± 0.77%) or mSOF (11.53 ± 1.86%). In case of with-zona parthenogenesis, RVCL medium and EDM were found superior to mSOF. The results of the study will be helpful to improve the efficiency of nuclear transfer in goat.