Activation of murine oocytes with Ca2+ ionophore and cycloheximide.
A Ca2+ ionophore (A23187, 3 microM) and inhibitor of protein synthesis (cycloheximide, 10 micrograms/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.
Department of Meat and Animal Science, University of Wisconsin, Madison 53706.
L J
LJ
Hagemann
F L
FL
Hillery-Weinhold
M L
ML
Leibfried Rutledge
N L
NL
First
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology*; D003513:Cycloheximide / Q000494:pharmacology*; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D010312:Parthenogenesis
D000001:Calcimycin; D003513:Cycloheximide