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. 2016 Sep 7;291(44):23257–23267. doi: 10.1074/jbc.M116.749747

FIGURE 4.

FIGURE 4.

PDE1 inhibition decreases D1R agonist-induced cAMP levels and GluA1 surface insertion by activating PDE2. Data are represented as mean ± S.E. normalized to baseline. A, PDE1 inhibition increases cGMP levels. 0.1 μm A68930 was added after 5 min of baseline, followed by DMSO (replotted from Fig. 3 for comparison) or 10 μm MMPX (black, 11 cells). *, p = 0.0373 (repeated measures two-way ANOVA, F1,17 = 5.11). B, PDE1 inhibition by MMPX decreases D1R agonist-induced cAMP levels. 0.1 μm A68930 was added for 10 min, followed by DMSO (replotted from Fig. 1 for comparison) or 10 μm MMPX (black, 14 cells) or 1.0 μm BAY60-7550 + 10 μm MMPX (black open squares, 11 cells). **, p = 0.0015 (repeated measures two-way ANOVA, DMSO versus MMPX, F1,29 = 12.39). ***, p < 0.0001 (repeated measures two-way ANOVA, MMPX versus MMPX + BAY60-7550, F1,22 = 25.61). C, PDE1 inhibition reduces GluA1 surface levels. The D1R agonist A68930 (0.1 μm) was added for 10 min, followed by DMSO (gray, 12 cells) or the PDE1 inhibitor MMPX (10 μm, black filled squares, 10 cells), or 1.0 μm BAY60-7550 + 10 μm MMPX (black open squares, 11 cells). **, p = 0.0016 (repeated measures two-way ANOVA, DMSO versus MMPX, F1,20 = 13.40). ***, p = 0.0005 (repeated measures two-way ANOVA, F1,19 = 17.29). D, expression of PDE2 mutants abolished PDE1 regulation of GluA1 surface insertion. Expression of PDE2 lacking catalytic activity (PDE2DN, gray squares, 15 cells) or allosteric regulation by cGMP (PDE2D485A, open circles; 14 cells) increases GluA1 surface insertion in the presence of 10 μm MMPX. ***, p < 0.0001 (repeated measures two-way ANOVA, MMPX versus PDE2DN, F1,23 = 27.70). ***, p = 0.0002 (repeated measures two-way ANOVA, MMPX versus PDE2D485A, F1,22 = 20.15).