Figure 1

Cyclic AMP-degrading PDE activity in the oocyte. A. Cyclic AMP-PDE activity was measured in the COC either alone or in the presence of a non-specific PDE inhibitor (IBMX, 1 mM) or of PDE3 (cilostamide, 10 μM) or PDE4 (rolipram, 10 μM) specific inhibitors. B. Total and calculated IBMX-sensitive, cilostamide-sensitive (PDE3) and rolipram-sensitive (PDE4) PDE activities in the COC are presented. To determine the inhibitor-sensitive contributions to the overall PDE activity, the activity measured in the presence of the respective inhibitors was subtracted from the total PDE activity. C. Comparison of the total (hatched bars) and cilostamide-sensitive (PDE3) (closed bars) PDE activities in the COC or DO after recovery from the ovary. A total of 1290 oocytes were used. Each column represents the mean ± SEM of at least three replicates. D. Reverse transcription-PCR analysis of β-actin and PDE3A mRNA in porcine oocyte cDNA. Ethidium bromide-stained 1% agarose gels were used to resolve the amplified products. DO: denuded oocyte cDNA; +ve: positive control; -ve: negative control without cDNA. A PDE3A amplicon in the oocytes was sequenced and found to have 98.3% identity with the published sequence. Representative pictures of the three different experiments are shown.