Analysis of caspase-3, caspase-8 and caspase-9 enzymatic activities in mouse oocytes and zygotes

Abstract

The current consensus in the literature is that ovulated oocytes that are not fertilized die by apoptosis, but the details of the proteins involved in the apoptotic pathways have not been elucidated. In this paper we confirm that caspase-3, the executioner of apoptosis, is expressed in mouse oocytes, and show that two initiators of apoptosis, caspase-8 and caspase-9, are expressed in mouse oocytes. Comparisons were made of caspase-3, -8, and -9 activities in superovulated oocytes that were freshly collected or allowed to age in vivo or in vitro. We found that caspase-3 activity significantly increased in aged oocytes compared with young oocytes (p<0.001), and that both caspase-8 activity and caspase-9 activity decreased in aged oocytes compared with young oocytes (p<0.001 for caspase-8 and p<0.05 for caspase-9 activity). A comparison of superovulated with naturally ovulated oocytes showed the same amount of caspase-8 activity in each, but a significant (p<0.001) decrease in caspase-9 activity in naturally ovulated compared with superovulated oocytes. There was no difference in caspase-3, -8, or -9 activity in oocytes compared with zygotes. Finally, we showed that culture of oocytes in staurosporine increased the activity of caspase-8 and caspase-9. In conclusion, the finding of both caspase-8 and caspase-9 activity in oocytes shows that unfertilized oocytes have the machinery to undergo apoptosis by using either the extrinsic (caspase-8 dependent) or intrinsic (caspase-9 dependent) pathways.