Fig 2: The effect of SCH23390 on the DPCPX-induced GluA1 S845 phosphorylation in the rat striatum. (a) Representative immunoblots illustrating the effect of SCH23390 on the DPCPX-induced S845 phosphorylation in the striatum. (b–d) Quantifications of the effect of SCH23390 on the DPCPX-induced S845 phosphorylation in the striatum. Note that SCH23390 reduced basal S845 phosphorylation and completely blocked the DPCPX-induced S845 phosphorylation. Rats were administered with SCH23390 (SCH, 0.1 mg/kg, i.p.) 10 min prior to DPCPX (2.5 mg/kg, i.p.) and were then sacrificed 15–20 min after DPCPX administration for subsequent analysis of changes in S845 and S831 phosphorylation using Western blots. Data are presented as means ± SEM (n = 4 per group) and were analyzed using two-way ANOVA: pS845: saline versus SCH23390, F(1,12) = 51.72, p < .001, vehicle versus DPCPX, F(1,12) = 11.99, p = .005, and interaction, F(1,12) = 7.17, p = .020. *p < .05 versus saline + vehicle. + p < .05 versus saline + DPCPX

Fig 3: The time‐dependent effect of DPCPX on GluA1 phosphorylation in the rat mPFC and hippocampus. (a) The time‐dependent effect of DPCPX on GluA1 phosphorylation in the mPFC. (b) The time‐dependent effect of DPCPX on GluA1 phosphorylation in the hippocampus. Note that DPCPX induced a biphasic effect in the mPFC, that is, an increase followed by a decrease in pS845 levels. Representative immunoblots are shown left to the quantified data. Rats received an i.p. injection of vehicle or DPCPX (2.5 mg/kg) and were then sacrificed at a different time point (1, 3, or 6 hr) after drug administration for subsequent analysis of changes in S845 and S831 phosphorylation using Western blots. Data are presented as means ± SEM (n = 4 per group) and were analyzed using Student's t test: mPFC‐pS845: p = .006 (1 hr), p = .022 (3 hr), and p = .522 (6 hr); and hippocampus‐pS845: p = .001 (1 hr), p = .040 (3 hr), and p = .220 (6 hr). *p < .05 versus vehicle at the same time point

Fig 4: GluA1 Ser845 phosphorylation levels are reversed with administration of NBI. Animals were separated into groups (A) balanced for anxiety-like behavior on the elevated plus maze (B) and received a post-conditioning infusion of either the CRFR1 antagonist NBI30775 or vehicle and an immediate extinction session 30 min later (C). Treatment with NBI30775 restored GluA1 Ser845 phosphorylation to vehicle home cage (HC) levels (D,E) but had no effect on GluA1 Ser831 (D,F) or on its own (G). Data depicted as percentage of vehicle HC control group (hatched bar and also dotted line) ± SEM. *Significant difference (p < 0.05) with the vehicle-treated HC group; n = 8–12/group.

Fig 5: CaMKII pT305/306 promotes LTD-induced GABAAR insertion at inhibitory synapses.Quantifications show means ± SEM. *P < 0.05 and **P < 0.01. Scale bars, 10 µm. (A) Schematic of CaMKII movement in response excitatory LTD stimuli. The LTD-induced pT305/306 blocks CaMKII movement to glutamatergic excitatory synapses and instead enables movement to GABAergic inhibitory synapses. (B) cLTD stimulation increased surface GABAAR accumulation in WT, but not T305/6AV, CA1 mini-slices (unpaired two-tailed t test, P = 0.0033 for WT and P = 0.6760 for AV, n = 5 and 6 samples (two slices/sample), as detected by immunoblot after surface biotinylation. (C) cLTD stimulation decreased surface GluA1 accumulation in WT, but not T305/6AV, CA1 mini-slices (unpaired two-tailed t test, P = 0.0062 for WT and P = 0.6252 for AV, n = 5 and 6 samples (two slices/sample). (D) cLTP decreased surface GABAAR cluster size in nonpermeabilized neurons from WT mouse hippocampal cultures (fixed 5 min after washout), while cLTD increased the cluster size (fixed 20 min after washout) (one-way ANOVA, Tukey’s post hoc test versus control, P = 0.0273 for cLTP and P = 0.0046 for cLTD; n = 12, 12, and 13 neurons), as detected by immunocytochemistry. (E) By contrast, in nonpermeabilized neurons from T305/5AV hippocampal cultures, cLTD did not increase surface GABAAR clusters. However, cLTP still decreased surface GABAAR clusters (P = 0.0147 for cLTP and P = 0.7121 for cLTD; n = 15, 13, and 14 neurons).