The means by which cumulus cells react to gonadotropin stimulation and regulate the subsequent production and degradation of cAMP are largely unknown. In this article we report that cyclic nucleotide phosphodiesterase type 3A (Pde3a) is transcriptionally regulated in porcine cumulus cells by a cAMP-dependent pathway during in vitro maturation (IVM). Cyclic AMP-PDE activity was increased in the cumulus-oocyte complex (COC) after 10 hours of IVM and 78% of this increase was sensitive to a Pde3-specific inhibitor, cilostamide. While no variation was observed in the oocyte, cilostamide-sensitive cAMP-PDE activity increased in the cumulus cells after IVM. This was supported by Western blotting, which showed that the intensity of a 135 kDa anti-Pde3a immunoreactive band was increased in COC after IVM. The Pde3a mRNA level was upregulated 28-fold in the COC after 4 hours of IVM and remained high up to 12 hours. The mRNA upregulation and increased activity were inhibited by an RNA synthesis inhibitor, -amanitin. The cilostamide-sensitive increase in PDE activity was inhibited by a protein synthesis inhibitor, cycloheximide. Pregnant mare serum gonadotropin (PMSG) caused dose-dependent activation of Pde3. The PMSG-dependent increase in Pde3 activity and Pde3a mRNA were mimicked by the adenylyl cyclase activator forskolin or prostaglandin E2. PMSG-dependent Pde3 activation was inhibited by the PKA-specific inhibitor, H89. Collectively, our results show for the first time that degradation of the intracellular cyclic nucleotide by PDE3A is transcriptionally upregulated via a cAMP-dependent pathway in cumulus cells, suggesting that it has a functional role during the ovulatory gonadotropin surge.