As expected from the cGMP FRET measurements, MRP4 inhibition increased the relaxation elicited by submaximal GS-NO concentrations clearly demonstrating that export of cGMP through MRP4 has an impact on cGMP-mediated vascular relaxation. In contrast, ANP- or CNP-stimulated cGMP signals near the EC50(ANP or CNP) were increased by PDE3 or PDE5 inhibition to a similar extent. How do isoproterenol-induced cAMP signals increase cGMP then? Impact of combined inhibition of cyclic GMP (cGMP) export and PDEs on S-nitroso-L-glutathione (GS-NO)–induced cGMP signals. According to the higher potency of ANP compared with CNP to relax blood vessels, 10-fold lower concentrations of ANP than of CNP were required to trigger cGMP signals. To prove the physiological relevance of these cGMP signals, relaxation of aortic rings was analyzed in organ bath experiments.
Thus, we analyzed the cGMP export by measuring intracellular and extracellular cGMP levels in radioimmunoassays. Impact of PDE3 and PDE5 on atrial natriuretic peptide (ANP)-induced cyclic GMP (cGMP) signals of primary smooth muscle cells. The findings indicate that cGMP export out of smooth muscle cells is a key player in the regulation of smooth muscle cGMP signals and blood vessel relaxation. Local Info In 2 previous studies analyzing smooth muscle cGMP signals with FRET-based cGMP sensors, DEA-NO was used to elicit cGMP signals. E, Smooth muscle cells were stimulated by 10 µmol/L MK-571, 1 µmol/L isoproterenol, or a combination thereof, and cGMP was measured by live cell microscopy using the FRET indicator cGi-6000.
Here, sildenafil almost abrogated the decline of cGMP during the following 9 minutes indicating that no other PDE was able to compensate for the missing PDE5 activity (Figure 2E and 2F, red traces). In the vascular system, cGMP lowers blood pressure by regulating vascular tone and endothelial permeability; furthermore, cGMP inhibits platelet aggregation.
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