Background and Aims: Previous studies have shown that caffeine might cause artificial dephosphorylation at threonine-14 and tyrosine-15 of the p34cdc2 catalytic subunit of maturation-promoting factor (MPF), elevate MPF activity in metaphase II oocytes cultured for a prolonged period, and that caffeine decreases fragmentation in oocytes cultured for up to 96 h. Methods: Studies were carried out on: (i) the effect of caffeine on the morphological status of oocytes cultured for 96 h; (ii) the parthenogenetic activation and the fertilization of oocytes incubated in a medium that contained caffeine, and (iii) the fertilization and preimplantation development ability of zona-intact and zona-free oocytes by in vitro fertilization (IVF) and by intracytoplasmic sperm injection. Results: In parthenogenetic activation, the incidence of diploid parthenotes in 24-h caffeine-treated oocytes was significantly higher than 24-h non-treated oocytes. For fertilizability of these oocytes, a significant increase in the fertilization rate resulted from IVF after 12-h caffeine incubation. Although no fertilized eggs were observed after intracytoplasmic sperm injection in 24-h non-treated oocytes, fertilized eggs were observed in caffeine-treated oocytes. MPF activation occurs in relation to nuclear/spindle position, and mitotic spindles and actin filaments determine the site of cleavage during cytokinesis. Spindle disruption does not cause cytofragmentation, but does induce cell cycle arrest. Conclusion: Based on our results, although caffeine might increase MPF activity, prolonged time in any incubation causes some disruption of cytoskeletal filaments, which might be responsible for the poor development of caffeine-treated oocytes. (Reprod Med Biol 2004; 3: 245-251).
Background and Aims: Previous studies have shown that caffeine might cause artificial dephosphorylation at threonine-14 and tyrosine-15 of the p34cdc2 catalytic subunit of maturation-promoting factor (MPF), elevate MPF activity in metaphase II oocytes cultured for a prolonged period, and that caffeine decreases fragmentation in oocytes cultured for up to 96 h. Methods: Studies were carried out on: (i) the effect of caffeine on the morphological status of oocytes cultured for 96 h; (ii) the parthenogenetic activation and the fertilization of oocytes incubated in a medium that contained caffeine, and (iii) the fertilization and preimplantation development ability of zona-intact and zona-free oocytes by in vitro fertilization (IVF) and by intracytoplasmic sperm injection. Results: In parthenogenetic activation, the incidence of diploid parthenotes in 24-h caffeine-treated oocytes was significantly higher than 24-h non-treated oocytes. For fertilizability of these oocytes, a significant increase in the fertilization rate resulted from IVF after 12-h caffeine incubation. Although no fertilized eggs were observed after intracytoplasmic sperm injection in 24-h non-treated oocytes, fertilized eggs were observed in caffeine-treated oocytes. MPF activation occurs in relation to nuclear/spindle position, and mitotic spindles and actin filaments determine the site of cleavage during cytokinesis. Spindle disruption does not cause cytofragmentation, but does induce cell cycle arrest. Conclusion: Based on our results, although caffeine might increase MPF activity, prolonged time in any incubation causes some disruption of cytoskeletal filaments, which might be responsible for the poor development of caffeine-treated oocytes. (Reprod Med Biol 2004; 3: 245-251).