Phosphodiesterase 5 Attenuates the Vasodilatory Response in Renovascular Hypertension
Figure 1
Reduced cGMP response in kidneys of NO-GC1 KO mice.
A, NO-stimulated cGMP-forming activity (100 µM DEA-NO) determined in kidney homogenates of WT and NO-GC1 KO mice (n=19 and 6 mice, respectively). PDE activity in WT and KO kidneys measured with 1 µM cGMP as substrate (n=11 and 7 mice, respectively). ** P< 0.01 versus WT, unpaired Student's t test. B, absent of the α1 subunit in NO-GC1 KO kidney homogenates and quantification of the α2 and β1 subunit content with respect to the subunit amount in WT using subunit specific antibodies (n=8 mice per genotype). Representative strips with α1, α2, β1 and the respective β tubulin bands of the same lanes are shown above. *** P< 0.001 versus WT, unpaired Student's t test. C, cyclic GMP content determined in renal cortical slices of WT and KO mice without any addition (n=20 and 10 mice, respectively), or in the presence of ODQ (20 µM, 15 min; n=5 and 4 mice, respectively) or carbachol (30 µM, 3 min; n=10 and 6 mice, respectively) or DEA-NO (100 µM 3 min; n=10 and 7 mice, respectively). # P< 0.01 versus untreated slices in the same group; *P< 0.01 versus WT, unpaired Student's t test. D, carbachol-induced relaxation of NA-contracted isolated perfused kidneys of NO-GC1 KO (n=12) and WT mice (n=21). ** P< 0.01 versus WT, unpaired Student's t test. E, concentration-response curves for GSNO-induced relaxation of NA-contracted isolated perfused kidneys in the presence of L-NAME (n=5 WT and 7 KO mice). The white dots indicate the Cch-induced relaxation plotted on the curves to determine the corresponding GSNO concentration. F, Western blot detection of eNOS in 50 µg kidney homogenates of NO-GC1 KO (n=8) and WT mice (n=5) and quantification with respect to the eNOS amount in WT kidneys. A representative strip with eNOS and the respective β tubulin bands of the same lanes is shown above. * P< 0.05 versus WT, unpaired Student's t test.