camp dependent protein kinase inhibitor  (Tocris)

Journal: The Journal of physiology

Article Title: HCN1 channels mediate mu opioid receptor long-term depression at insular cortex inputs to the dorsal striatum

doi: 10.1113/JP283513

Figure Lengend Snippet: A, representative eEPSC traces showing the effects of DAMGO (0.3 μM, 5 min) application after the preincubation of PKA-selective inhibitor, KT5720 (1 μM, ≥1 hr). B–C, the preincubation of KT5720 blocked glutamatergic MOR-LTD, the eEPSC amplitudes did not change after DAMGO application (0–10 min baseline vs. final 10 min of recording; paired t test, P = 0.865, t7 = 0.176, n = 8 neurons from three mice). D, representative eEPSC traces showing the effects of DAMGO (0.3 μM, 5 min) application after the intracellular dialysis of PKI (1 μM, ≥30 min). E–F, the inhibition of postsynaptic PKA did not alter MOR-LTD. eEPSC amplitudes were reduced after DAMGO application (0–10 min baseline vs. final 10 min of recording; paired t test, P = 0.000120, t10 = 6.07, n = 11 neurons from five mice). G, presynaptic PKA inhibition disrupted MOR-LTD (KT5720: 101 ± 7%, P = 0.000193 vs. PKI: 84 ± 2%, P = 0.121; F(2,26) = 10.65, one-way ANOVA Dunnet’s multiple comparison test). H, representative oEPSC traces showing the effects of DAMGO (0.3 μM, 5 min) application after the preincubation of PKA-selective inhibitor, KT5720 (1 μM, ≥1 h). I–J, PKA inhibition blocked corticostriatal MOR-LTD, with no changes in oEPSC amplitudes after DAMGO application (0–10 min baseline vs. final 10 min of recording; paired t test, P = 0.183, t8 = 1.46, n = 9 neurons from four mice). K, representative oEPSC traces showing the effects of DAMGO (0.3 μM, 5 min) application after the intracellular dialysis of PKI (1 μM, ≥30 min). L–M, the inhibition of postsynaptic PKA did not alter MOR-LTD. oEPSC amplitudes were reduced after DAMGO application (0–10 min baseline vs. final 10 min of recording; paired t test, P = 0.000118, t9 = 6.45, n = 10 neurons from four mice). N, presynaptic PKA inhibition disrupted cortical MOR-LTD (KT5720: 94 ± 5%, P = 0.0305 vs. PKI: 64 ± 6%, P = 0.235; F(2,25) = 8.805, One-way ANOVA Dunnet’s multiple comparison test). O, representative AIC-DLS oEPSC traces showing the effects of DAMGO (0.3 μM, 5 min) application after the preincubation of PKA-selective inhibitor, KT5720 (1 μM, ≥1 h). P–Q, KT5720 blocked AIC-expressed MOR-LTD, with no changes in oEPSC amplitudes after DAMGO application (0–10 min baseline vs. final 10 min of recording; paired t test, P = 0.141, t9 = 1.613, n = 10 neurons from three mice). R, representative AIC-DLS oEPSC traces showing the effects of DAMGO (0.3 μM, 5 min) application after the intracellular dialysis of PKI (1 μM, ≥30 min). S–T, postsynaptic PKA inhibition did not alter specific AIC MOR-mediated LTD. oEPSC amplitudes were reduced after DAMGO application (0–10 min baseline vs. final 10 min of recording; paired t test, P = 0.00488, t5 = 4.8, n = 6 neurons from three mice). U, presynaptic PKA inhibition disrupted AIC MOR-LTD (KT5720: 94 ± 4%, P = 0.0171 vs. PKI: 69 ± 5%, P = 0.518; F(2,20) = 8.411, one-way ANOVA Dunnet’s multiple comparison test). Time course data represent means ± SEM. Box plots show average of the final 10 min of recording and represent median and interquartile ranges. ns = not significant, **P<0.01, ***P<0.001.

Article Snippet: Reagents We used the MOR agonist [D-Ala 2 , NMe-Phe 4 , Gly-ol 5 ]-enkephalin (DAMGO; H-2535, Bachem), GABA A receptor antagonist picrotoxin (PTX, P1675, Sigma-Aldrich), PKI (6221, Tocris), KT5720 (1288, Tocris), Forskolin (F3917, Sigma-Aldrich), Cyclo-heximide (0970, Tocris), ZD7288 (1000, Tocris), WIN55,212-2 (1038, Tocris) and Torin 2 (4248, Tocris).

Techniques: Inhibition, Comparison

Journal: The Journal of physiology

Article Title: HCN1 channels mediate mu opioid receptor long-term depression at insular cortex inputs to the dorsal striatum

doi: 10.1113/JP283513

Figure Lengend Snippet: A, representative optically evoked EPSC traces before, during and after DAMGO (0.3 μM, 5 min) application in brain slices from VGluT2-Ai32 mice. B–C, thalamocortical inputs produced MOR-STD in DLS from VGluT2-Ai32 mice (10 min baseline vs. peak 12–20 min of recording P = 0.00275; 12–20 min Peak vs. 30–35 min Wash out of recording, P = 0.0889, repeated measure one-way ANOVA Tukey’s test, n = 7 neurons from three mice). D, representative oEPSC traces showing the effects of DAMGO (0.3 μM, 5 min) application after the preincubation of PKA-selective inhibitor KT5720 (1 μM, ≥1 h, blue open square) and after the intracellular dialysis of PKI (1 μM, ≥30 min, cyan open square). E–F, the inhibition of pre (1 μM KT5720) and postsynaptic PKA (1 μM PKI) did not affected MOR-STD (KT5720: 10 min baseline vs. peak 12–20 min of recording, P = 0.0353; 12–20 min Peak vs. 30–35 min Wash out of recording, P = 0.0353, repeated measure one-way ANOVA Tukey’s test, n = 7 neurons from three mice) (PKI: 10 min baseline vs. peak 12–20 min of recording, P = 0.00218; 12–20 min Peak vs. 30–35 min Wash out of recording, P = 0.0429, repeated measure one-way ANOVA Tukey’s test, n = 9 neurons from three mice). G, representative oEPSC traces showing the effects of 20 μM forskolin before, during and after DAMGO (0.3 μM, 5 min) application in brain slices from VGluT2-Ai32 mice. H–I, MOR-STD was unaffected after the activation of AC by 20 μM forskolin (10 min baseline vs. peak 12–20 min of recording, P = 0.00260; 12–20 min Peak vs. 30–35 min Wash out of recording, P = 0.168, repeated measure one-way ANOVA Tukey’s test, n = 8 neurons from four mice). J, representative oEPSC traces showing the effects of 80 μM cycloheximide before, during and after DAMGO (0.3 μM, 5 min) application in brain slices from VGluT2-Ai32 mice. K–L, protein translation is not necessary for the expression of MOR-STD (10 min baseline vs. peak 12–20 min of recording, P = 0.00973; 12–20 min Peak vs. 30–35 min Wash out of recording, P = 0.0822, repeated measure one-way ANOVA Tukey’s test, n = 7 neurons from three mice). M, the box plot illustrates the average peak current (12–20 min) and summarize the findings on MOR-STD mechanism in the DLS (one-way ANOVA Dunnett’s multiple comparisons test). Time course data represent means ± SEM. Box plots represent median and interquartile ranges and show average peak responses (12–20 min). ns = not significant, *P < 0.05, **P < 0.01.

Article Snippet: Reagents We used the MOR agonist [D-Ala 2 , NMe-Phe 4 , Gly-ol 5 ]-enkephalin (DAMGO; H-2535, Bachem), GABA A receptor antagonist picrotoxin (PTX, P1675, Sigma-Aldrich), PKI (6221, Tocris), KT5720 (1288, Tocris), Forskolin (F3917, Sigma-Aldrich), Cyclo-heximide (0970, Tocris), ZD7288 (1000, Tocris), WIN55,212-2 (1038, Tocris) and Torin 2 (4248, Tocris).

Techniques: Produced, Inhibition, Activation Assay, Expressing