
%0 Journal Article
%A Bali Papp, A. 
%A Varga, E. 
%T 312 CHEMICAL ACTIVATION OF <i>IN VITRO</i>-MATURED PORCINE OOCYTES
%J Reproduction, Fertility and Development
%D 2005/12/14
%V 18
%N 2
%P 263-264
%K 
%R https://doi.org/10.1071/RDV18N2AB312
%U https://www.publish.csiro.au/paper/RDV18N2AB312 
%X Parthenogenetic oocyte activation is important for nuclear transfer and for the understanding of cell cycle regulation of oocytes. Several chemical agents, including ethanol, cycloheximide, strontium, cytochalasin B, 6 dimethylaminopurine, CaCl2 and ionophore A23187 can induce mammalian oocyte activation in vitro. The objectives of the present study were: (1) to assess the ability of strontium chloride (S), cytochalasin B (CB), cycloheximide (CX), and 6-dimethylaminopurine (D) to induce activation and parthenogenetic development in porcine oocytes; and (2) to verify whether the combinations of treatments (SB group = strontium combined with cytochalasin; SX group = strontium combined with cycloheximide, and SD group = strontium combined with 6-dimethylaminopurine) improves activation and parthenogenetic development rates. Oocytes from slaughterhouse ovaries were matured in vitro for 42 h at 39°C, in 5% CO2 in air. The basic medium used for oocyte maturation was TCM-199 supplemented with 10% pig follicular fluid, 1.25 mM L-glutamine, 0.9 mM Na-pyruvate, 100 ¼M cysteamine, 0.1 mg/mL streptomycin sulfate, 10 IU/mL pregnant mare serum gonadotropin (PMSG), and 10 IU/mL hCG (Werfft-Chemie GmbH, Vienna, Austria). Denuded MII oocytes were cultured in activation solution for 5 h. Thereafter the oocytes were cultured in NCSU37 for 6 days. At 48 h and 6 days after activation, oocytes, zygotes were fixed in acetic acid:alcohol (1/3 w/v), then stained with 0.1% (w/v) orcein in 45% (v/v) acetic acid, and evaluated under a phase contrast microscope. Each experiment was repeated four times. All data were analyzed by ANOVA, followed by Duncan's multiple range test (P < 0.05). A total of 2243 oocytes were activated in the different groups. In all groups, more than 45% of the oocytes were activated. No significant difference was observed in activation rate among SD (346/170, 49.13%), SX (302/164, 54.3%), and SB (318/182, 57.23%) groups. The activation rate for CB was significantly higher (P < 0.05) than for D or S (323/192, 59.44 ± 6.84%; 366/176, 48.09 ± 3.43%; and 319/183, 53.29 ± 5.39%, respectively). The blastocyst rate for SX was significantly higher (P < 0.05) than that for D, SD, or SB (8.64 ± 8.07%; and 0 ± 0%; 0 ± 0%; and 1.27 ± 2.41%, respectively). In conclusion, this study suggests that chemical activation procedure is the most effective in strontium chloride combined with cycloheximide. The lowest oocyte fragmentation rates were in SX (28.40 ± 1.26%) and CX (21.05 ± 1.12%). This work was supported by the the Hungarian Scientific T 43131 Research Foundation and the Hungarian Science on Technology Foundation E 14/04. 
