ACTIVATION OF MURINE OOCYTES WITH CA2+ IONOPHORE AND CYCLOHEXIMIDE

A Ca2+ ionophore (A23187, 3 mu M) and inhibitor of protein synthesis (cycloheximide, 10 mu g/ml) were used sequentially as a unique method for activating mouse oocytes in vitro. Brief exposure of oocytes to A23187 followed by 6 hr in cycloheximide resulted in a higher activation rate (93.8%) compared to A23187 or cycloheximide alone (37.7% and 36.5%, respectively) or the two reagents in reverse order (29.8%). The parthenogenones consistently contained a single pronucleus and second polar body, and showed a high degree of developmental potential, as assessed by transfer to recipient females or addition of a male pronucleus followed by transfer to recipients. This method is a useful way of obtaining large numbers of activated haploid mammalian oocytes for further developmental studies. (C) 1995 Wiley-Liss, Inc.