%0 Journal Article %A Tomas C. Bellamy %A John Garthwaite %T “cAMP-Specific” Phosphodiesterase Contributes to cGMP Degradation in Cerebellar Cells Exposed to Nitric Oxide %D 2001 %R 10.1124/mol.59.1.54 %J Molecular Pharmacology %P 54-61 %V 59 %N 1 %X Nitric oxide (NO) functions as a diffusible messenger in the central nervous system and elsewhere, exerting many of it physiological effects by activating soluble guanylyl cyclase, so increasing cellular cGMP levels. Hydrolysis of cyclic nucleotides is achieved by phosphodiesterases (PDEs) but the enzyme isoforms responsible for degrading cGMP in most cells have not been identified. We have devised a method for quantitatively monitoring the rate of breakdown of cGMP within intact cells and have applied it to rat cerebellar cell suspensions previously stimulated with NO. In contrast to previous findings in cultured cerebellar cells, there was no evidence from the use of selective inhibitors that PDE 1 participated importantly in cGMP hydrolysis. Moreover, procedures expected to increase PDE 1 activity by raising cytosolic Ca2+ concentrations (neurotransmitter agonists, Ca2+ ionophore) failed to influence cGMP breakdown. Instead, through the use of inhibitors selective for different PDE families, two isoforms were implicated: a “cGMP-specific” PDE (PDE 5), inhibited by sildenafil and zaprinast, and a “cAMP-specific” PDE (PDE 4), inhibited by low concentrations of rolipram and Ro-20–1724 and by milrinone. An explanation is offered for a participation of PDE 4 based on the high estimated intracellular cGMP concentration (∼800 μM) and the low affinity of the enzyme for cGMP. In accordance with predictions, recombinant PDE 4 was shown to hydrolyze high cGMP concentrations in a rolipram-sensitive manner. The widespread use of rolipram to test for a specific involvement of cAMP in cellular phenomena must therefore be questioned. %U https://molpharm.aspetjournals.org/content/molpharm/59/1/54.full.pdf