1. Imbalanced Activity in the Orbitofrontal Cortex and Nucleus Accumbens Impairs Behavioral Inhibition 
Heidi C. Meyer, David J. Bucci 
Current Biology 
Volume 26, Issue 20, Pages (October 2016)
DOI: /j.cub
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2. Figure 1
Configuration of Stimuli in the Negative Occasion Setting Procedure and Schematic of the Balance Model of Behavioral Control
(A) Red and green lines indicate inhibitory and excitatory relationships in the behavioral procedure, respectively (US, unconditioned stimulus). The feature stimulus acts to gate or set the occasion for the meaning of the target stimulus and indicates that a response should be withheld during the subsequent presentation of the target [21].
(B) Balanced activity (left side of panel, and indicated by dashed line) is necessary for appropriate behavioral control and is present in preadolescence and adulthood. An imbalance can result from impairing the function of OFC (middle left [7]), increasing the influence of NAC (middle right [8]), or by simultaneously disrupting and potentiating activity in OFC and NAC, respectively [2, 5, 9, 10].
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3. Figure 2
Example of Conditioned Food Cup Behavior during Training in the Negative Occasion Setting Procedure
(A) Conditioned responding during presentation of the tone on reinforced (R) and non-reinforced trials (NR) and (B) the difference in responding between trial types, exhibited by the vehicle-treated control group in experiment 1. Asterisk refers to the first of three consecutive sessions during which the group’s difference score between R and NR trials was significantly different from zero, defined as a Z score ≥ (i.e., p < 0.01; see Supplemental Experimental Procedures). Data are means ± SEM.
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4. Figure 3
Summary Data Indicating the Number of Sessions until Each Group in the Study Consistently Exhibited a Significant Difference in Responding to the Tone on Reinforced and Non-reinforced Trials
Significant difference is defined as Z score ≥ 2.325, p < 0.01 for three consecutive sessions; see Supplemental Experimental Procedures. Data in black are from experiment 1 (combined manipulations of OFC and NAC), data in blue are from experiment 2 (manipulation of OFC alone), and data in red are from experiment 3 (manipulation of NAC alone). Rats with combined manipulation of OFC and NAC required more training to consistently exhibit a significant difference in responding to the tone on reinforced versus non-reinforced trials compared to either manipulation alone (see Results). Dotted line indicates the mean number of sessions for all control groups in the study (the individual control groups in each experiment exhibited comparable learning (see Results); thus, only the controls that received the DREADDs virus and vehicle treatment are shown). All groups in experiments 1 and 2 exhibited low levels of baseline responding during the Pre-CS epoch (Ps > 0.3; see Supplemental Experimental Procedures). In experiment 3 there was a significant group difference [F(3,40) = 4.74, p < 0.01] in that the NAC-hM3Dq-CNO group had higher levels of baseline responding than rats in the NAC-GFP-CNO group, but there were no differences compared to either NAC-hM3Dq-vehicle or NAC-GFP-vehicle rats. Similarly high levels of feeding behavior during the Post-CS epoch were observed each experiment (Ps > 0.1). There was no effect of group on time spent rearing during the light in any experiment (Ps > 0.1). In addition, food cup behavior during the light was low (on average ∼0.75 s) and did not differ between groups in experiments 1 or 2 (P’s > 0.7). In experiment 3, a main effect of group was observed [F(3,40) = 4.24, p < 0.05], which was driven by lower levels of responding by rats in the NAC-GFP-CNO group relative to rats in the NAC-hM3Dq-CNO and NAC-hM3Dq-VEH groups and thus cannot explain the primary impairment observed in the NAC-hM3Dq-CNO group. Furthermore, although all groups showed a slight increase in responding during the first few sessions (likely attributable to a decrease in rearing during these same sessions), we did not observe any other changes in food cup responding during the light across training. OFC, orbitofrontal cortex; NAC, nucleus accumbens; CNO, treatment with clozapine-N-oxide; VEH, vehicle-treated control group.
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5. Figure 4 Expression of the DREADDs Reporter in OFC and NAC
(A and C) Fluorescent labeling of OFC neurons expressing hM4Di (A) and NAC neurons expressing hM3Dq (C) (20× magnification).
(B and D) Schematic diagram of representative minimum (dark gray) and maximum (light gray) expression of hM4Di in OFC (B) and hM3Dq in NAC (D). Expression of the reporter was comparable and evident along the rostrocaudal extent of each region. Few fluorescent neurons were observed outside of the target region. Expression of the reporter in the GFP control groups was comparable to that observed in the hM4Di groups (data not shown). Expression of the reporters for the viruses was very similar across experiments.
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