Parthenogenetic activation of rat oocytes and their development in vitro

Abstract

The present study was performed to determine suitable methods for parthenogenetic activation and subsequent development of rat oocytes in vitro. In the first series of experiments, the ability of electrical pulses, strontium, ethanol and ionomycin to activate Sprague-Dawley (SD) rat oocytes was examined. The synergistic effect of strontium and cycloheximide or puromycin was also examined in the second series of experiments. In the third series of experiments, the development of F1 hybrid (SD × Dark Agouti) parthenotes activated with different concentrations of strontium (10–0.08 mM) was compared with that of SD parthenotes. The effect of the timing of activation (10 min and 2, 4 and 6 h after cervical dislocation) was also assessed in a fourth series of experiments. The oocytes activated by strontium showed higher pronuclear formation and cleavage rates than those in the other groups (P < 0.05). Higher blastocyst development was obtained from parthenotes activated by strontium and strontium–cycloheximide compared with the strontium–puromycin group (P < 0.01). However, the total cell number of blastocysts from the strontium–cycloheximide activation group was higher than that of other groups (P < 0.05). With strontium (2.5–10 mM) treatment, 40.9% of blastocysts were obtained from F1 hybrid oocytes, whereas 22.9% were obtained from SD (P < 0.01). The oocytes activated 10 min or 2 h following cervical dislocation showed higher blastocyst development than those of the 4 and 6 h groups (P < 0.01). These results suggest that strontium–cycloheximide produces the highest parthenogenetic activation rate in the rat and that oocytes must be activated by 2 h after cervical dislocation.