Journal: Neuropsychopharmacology

Article Title: Frontostriatal regulation of brain circuits contributes to flexible decision making

doi: 10.1038/s41386-025-02065-8

Figure Lengend Snippet: A Experimental design. Rats were bilaterally injected with adeno-associated virus (AAV) expressing the excitatory DREADD hM3Dq or green fluorescent protein (GFP) as control (AAV5-syn-GFP) in the medial prefrontal cortex (mPFC) and bilateral cannulas were implanted 1 mm above the nucleus accumbens (NAc). After 2 weeks of recovery, rats underwent food pretraining for 5 days, visual-cue task (VCT) training for 1–2 days and response discrimination task (RDT) for 1 day. Rats were injected either with CNO or VEH into the NAc 20 min prior to the RDT. Five to seven days later, the fMRI experiment was conducted. In the scanner, rats underwent a baseline resting-state fMRI (rs-fMRI) scan to assess functional connectivity, followed by a CNO or VEH injection into the NAc. After 20 min, a post-injection rs-fMRI scan was performed followed by cerebral blood volume (CBV) to evaluate neural activity. The entire sequence was repeated 5–7 days later with reversed VEH or CNO injection order. Lastly, the rats were sacrificed for Immunofluorescence staining. B Example of VCT. The rat was required to respond on the lever that had a visual-cue stimulus light illuminated above it. C Example of RDT. The rat was required to cease the use of a visual-cue strategy, and instead use a spatial discrimination strategy to obtain food pellets. In this example, the rat was required to always press the left lever during the RDT. D Example of different subtypes of errors. The trials were separated into consecutive blocks of 8 trials each. A perseverative error was scored when the rat pressed the right lever when the light stimulus was on the right side in early stage of the choice sequence. However, if the proportion of such errors was less than 75% in a block (6 out of 8 trials), from that time point the error was scored as a regressive error . A never-reinforced error was scored when the rat chose the right lever without cue illuminated above it.

Article Snippet: Rats were bilaterally injected with adeno-associated virus (AAV) expressing the excitatory DREADD hM3Dq (AAV5-syn-hM3D-HA; Addgene, Watertown, MA) or green fluorescent protein (GFP) as control (AAV5-syn-GFP; Addgene) in the mPFC (AP +3.4 mm, ML ±0.7 mm, DV −3.8 mm) followed by cannulas placement 1 mm above the NAc (AP +1.5 mm, ML ±2.6 mm, DV −7 mm, angle 10°).

Techniques: Injection, Virus, Expressing, Control, Functional Assay, Activity Assay, Sequencing, Immunofluorescence, Staining, Blocking Assay

Journal: Neuropsychopharmacology

Article Title: Frontostriatal regulation of brain circuits contributes to flexible decision making

doi: 10.1038/s41386-025-02065-8

Figure Lengend Snippet: A There was no significant difference in the pretraining phase between the hM3Dq ( n = 19) and control ( n = 19) groups. B There was no significant difference in number of total trails to reach criterion between the two groups in VCT training. C Activation of the mPFC-NAc pathway significantly reduced the number of total trials needed to reach the criterion in RDT ( n = 9–10 per group). Data were analyzed by two-way ANOVA followed by Bonferroni post hoc test. * p < 0.05, hM3Dq-CNO vs. control-CNO; ** p < 0.01, hM3Dq-CNO vs. hM3Dq-VEH. D Activation of the mPFC-NAc pathway significantly increased the percentage of correct trials in RDT ( n = 9–10 per group). Data were analyzed by two-way ANOVA followed by Bonferroni post hoc test. * p < 0.05, hM3Dq-CNO vs. hM3Dq-VEH. E Activation of the mPFC-NAc pathway significantly reduced the number of incorrect trials in RDT ( n = 9–10 per group). Data were analyzed by two-way ANOVA followed by Bonferroni post hoc test. * p < 0.05, hM3Dq-CNO vs. control-CNO; ** p < 0.01, hM3Dq-CNO vs. hM3Dq-VEH. F Activation of the mPFC-NAc pathway significantly decreased perseverative errors in the hM3Dq group but not the control group. Two-way ANOVA followed by Bonferroni post hoc test. * p < 0.05, hM3Dq-CNO vs. hM3Dq-VEH. G Activation of the mPFC-NAc pathway had no effects on regressive errors. H Activation of the mPFC-NAc pathway had no effects on never-reinforced errors. I The comparison of cerebral blood volume (CBV) maps between VEH and CNO injections in the hM3Dq group ( n = 5). Data were analyzed by voxel-wise t -test. Results were corrected for whole-brain multiple comparisons at p corr < 0.05. J CNO injection in the NAc increased CBV in the orbitofrontal cortex (OFC), mPFC, piriform cortex (Pir), cingulate cortex (CG), striatum (STR), thalamus (THAL), hypothalamus (HYPO), retrosplenial dysgranular/granular cortex (RSD/RSG), parietal association cortex (PtA), visual cortex (VIS), hippocampus (HIP), auditory cortex (AU) and superior colliculus (SC). Coordinates represent distance relative to bregma (in millimeters). Error bars in the figures represent the SEM. VEH vehicle, CNO clozapine N-oxide.

Article Snippet: Rats were bilaterally injected with adeno-associated virus (AAV) expressing the excitatory DREADD hM3Dq (AAV5-syn-hM3D-HA; Addgene, Watertown, MA) or green fluorescent protein (GFP) as control (AAV5-syn-GFP; Addgene) in the mPFC (AP +3.4 mm, ML ±0.7 mm, DV −3.8 mm) followed by cannulas placement 1 mm above the NAc (AP +1.5 mm, ML ±2.6 mm, DV −7 mm, angle 10°).

Techniques: Control, Activation Assay, Comparison, Injection

Journal: Neuropsychopharmacology

Article Title: Frontostriatal regulation of brain circuits contributes to flexible decision making

doi: 10.1038/s41386-025-02065-8

Figure Lengend Snippet: A The top shows the location of NAc seed, which was defined according to the cannula tip in each rat. SESSION (baseline, post-injection) × GROUP (hM3Dq, control) × TREATMENT (CNO, VEH) ANOVA showed a significant interaction effect in functional connectivity of NAc with orbitofrontal cortex (OFC), olfactory bulb (OB), secondary motor cortex (M2), mPFC and sensory cortex (SEN). Results were corrected for whole-brain multiple comparisons at p corr < 0.05. B GROUP × TREATMENT ANOVA showed a significant interaction effect in functional connectivity of NAc with M2, mPFC and SEN after injection. However, there was no significant effect in the baseline. Results were corrected for whole-brain multiple comparisons at p corr < 0.05. C Post hoc analyses found a significant difference in functional connectivity of NAc with mPFC and M2 between rats with CNO or VEH injection in the hM3Dq group but not the control group. Results were corrected for whole-brain multiple comparisons at p corr < 0.05. D CNO injection decreased functional connectivity of NAc-mPFC in the hM3Dq group but not the control group. E Functional connectivity of NAc-mPFC was positively correlated with number of total trials to reach the criterion. Statistical significance was preset at p uncorr < 0.025 for multiple comparisons correction. F CNO injection decreased functional connectivity of NAc-M2 in the hM3Dq group but not the control group. G Functional connectivity of NAc-M2 was positively correlated with number of total trials to reach the criterion. Statistical significance was preset at p uncorr < 0.025 for multiple comparisons correction. N = 10 per group. Error bars in the figures represent the SEM. Coordinates represent distance relative to bregma (in millimeters). VEH vehicle, CNO clozapine N-oxide.

Article Snippet: Rats were bilaterally injected with adeno-associated virus (AAV) expressing the excitatory DREADD hM3Dq (AAV5-syn-hM3D-HA; Addgene, Watertown, MA) or green fluorescent protein (GFP) as control (AAV5-syn-GFP; Addgene) in the mPFC (AP +3.4 mm, ML ±0.7 mm, DV −3.8 mm) followed by cannulas placement 1 mm above the NAc (AP +1.5 mm, ML ±2.6 mm, DV −7 mm, angle 10°).

Techniques: Injection, Control, Functional Assay

Journal: Neuropsychopharmacology

Article Title: Frontostriatal regulation of brain circuits contributes to flexible decision making

doi: 10.1038/s41386-025-02065-8

Figure Lengend Snippet: A The top showed mPFC seed, which was defined as the region showing significance in the analysis on functional connectivity with the NAc seed. SESSION (baseline, post-injection) × GROUP (hM3Dq, control) × TREATMENT (CNO, VEH) ANOVA showed a significant interaction effect in functional connectivity of mPFC with frontal association cortex (FrA), secondary motor cortex (M2), orbitofrontal cortex (OFC), mPFC, cingulate cortex (CG), striatum (STR), septum (SEP), sensory cortex (SEN), bed nucleus of the stria terminalis(BNST), retrosplenial dysgranular cortex (RSD), parietal association cortex (PtA), visual cortex (VIS) and auditory cortex (AU). Results were corrected for whole-brain multiple comparisons at p corr < 0.05. B GROUP × TREATMENT ANOVA showed a significant interaction effect in functional connectivity of mPFC with FrA, OFC, M2, mPFC, CG, STR, SEP, SEN, RSD, PtA, VIS and AU after injection. However, there was no significant effect in the baseline. Results were corrected for whole-brain multiple comparisons at p corr < 0.05. C Post hoc analyses showed significant differences in functional connectivity of mPFC with FrA, OFC, M2, CG, STR, SEP, SEN, RSD, PtA, VIS and AU between rats with CNO or VEH injection in the hM3Dq group but not the control group. Results were corrected for whole-brain multiple comparisons at p corr < 0.05. D Chemogenetic activation of mPFC-NAc pathway decreased functional connectivity of the above mPFC-related circuits. E Loadings of mPFC-related circuits on the two PCA dimensions were clustered into three networks. F Functional connectivity of the Network 1, 2 and 3 was not related to number of total trials to reach criterion. Statistical significance was preset at p uncorr < 0.017 for multiple comparisons correction. N = 10 per group. Coordinates represent distance relative to bregma (in millimeters). VEH vehicle, CNO clozapine N-oxide.

Article Snippet: Rats were bilaterally injected with adeno-associated virus (AAV) expressing the excitatory DREADD hM3Dq (AAV5-syn-hM3D-HA; Addgene, Watertown, MA) or green fluorescent protein (GFP) as control (AAV5-syn-GFP; Addgene) in the mPFC (AP +3.4 mm, ML ±0.7 mm, DV −3.8 mm) followed by cannulas placement 1 mm above the NAc (AP +1.5 mm, ML ±2.6 mm, DV −7 mm, angle 10°).

Techniques: Functional Assay, Injection, Control, Activation Assay

Journal: Neuropsychopharmacology

Article Title: Frontostriatal regulation of brain circuits contributes to flexible decision making

doi: 10.1038/s41386-025-02065-8

Figure Lengend Snippet: A The top showed M2 seed, which was defined as the region showing significance in the analysis on functional connectivity with the NAc seed. SESSION (baseline, post-injection) × GROUP (hM3Dq, control) × TREATMENT (CNO, VEH) ANOVA showed a significant interaction effect in functional connectivity of M2 with orbitofrontal cortex (OFC), insula (INS), cingulate cortex (CG), striatum (STR), sensory cortex (SEN), M2, hippocampus (HIP) and auditory cortex (AU). Results were corrected for whole-brain multiple comparisons at p corr < 0.05. B GROUP × TREATMENT ANOVA showed a significant interaction effect on functional connectivity of M2 with OFC, INS, mPFC, CG, STR, SEN and M2 after injection. However, there was no significant effect in the baseline. Results were corrected for whole-brain multiple comparisons at p corr < 0.05. C Post hoc analyses showed significant differences in functional connectivity of M2 with OFC, INS, mPFC, CG, STR, SEN and M2 between rats with CNO or VEH injection in the hM3Dq group but not the control group. Results were corrected for whole-brain multiple comparisons at p corr < 0.05. D Chemogenetic activation of mPFC-NAc pathway decreased functional connectivity of the above M2-related circuits. E Loadings of M2-related circuits on the PCA dimension were clustered into two networks. F Functional connectivity of Network 1 but not Network 2 significantly correlated with number of total trials to reach criterion. Statistical significance was preset at p uncorr < 0.025 for multiple comparisons correction. N = 10 per group. Coordinates represent distance relative to bregma (in millimeters). VEH vehicle, CNO clozapine N-oxide.

Article Snippet: Rats were bilaterally injected with adeno-associated virus (AAV) expressing the excitatory DREADD hM3Dq (AAV5-syn-hM3D-HA; Addgene, Watertown, MA) or green fluorescent protein (GFP) as control (AAV5-syn-GFP; Addgene) in the mPFC (AP +3.4 mm, ML ±0.7 mm, DV −3.8 mm) followed by cannulas placement 1 mm above the NAc (AP +1.5 mm, ML ±2.6 mm, DV −7 mm, angle 10°).

Techniques: Functional Assay, Injection, Control, Activation Assay