Rats were injected with individual cytokines [CCL2 = chemokine (C-C motif) ligand 2; IL1β = interleukin-1β; TNFα = tumor necrosis factor-α] at a weekly interval prior to exposure to 5 days of 4.5% alcohol diet (ED). CD VEH = vehicle during control diet; ED-Veh = vehicle prior to alcohol diet. CD-CCL2 = CCL2 given weekly during control diet exposure. Social interaction as a measure of anxiety-like behavior was determined 5–6 hrs after alcohol removal. * P< 0.001 compared to CD-Veh and ED-Veh groups. From Breese et al., (2008).

P-rats were given either continuous voluntary access to water and alcohol solution (10%) or 3 cycles of voluntary access to water and alcohol solution with 2 days of abstinence between the 1^st two cycles. Intake of alcohol was subtracted from intake on the final 5 days of fluid ingestion.
*Significantly different from Continuous. +Significantly different from Cycling Only. Modified from Breese et al. (2004) by permission of the publisher.

Twenty-eight recently abstinent alcohol dependent (AD) and 24 healthy social drinkers (SD) are included in the trials. Whole brain voxel-based mixed effects model analyses for significant Group 2 (AD, SD) main effects and Group X Condition 3 (S, C, N) interaction effects are shown for the selected brain regions at specified thresholds of significance (p<.05, p<.01, p<.005 as noted). Talaraich coordinates (x,y,z) designate the center of activation in each brain region. Whole brain voxel based maps for main and interaction effects are followed by group difference maps (AD minus SD) depicted in the right three images for each test condition (Neutral, Stress, Alcohol Cue).
For (a), the nucleus accumbens/ventral striatum (NAc/VS) region, a significant group main effect is shown at two separate thresholds ([i] p<.05; [ii] p<.01 on the left in red/yellow). Simple effect contrasts show greater activation in AD subjects than SD controls in the neutral and stress conditions (p<.01; depicted in red/yellow), but not in the alcohol cue condition (i.e., the three images shown on the right). For (b), the orbitofrontal cortex (OFC), a significant group main effect was seen at two separate thresholds ([i] p<.05; [ii] p<.01 shown on the left). Simple effects to explain this significant group effect in the OFC indicated lower activity in AD patients relative to SD controls as shown in pink for the OFC region across all three conditions (p<.01; AD less than SD).
For (c), the left insula region, the Group X Condition significant interaction effect is shown on the left in red/yellow at [i] p<.01 and [ii] p<.005 thresholds. Simple effect contrasts to explain the significant interaction effect indicated greater activity in AD patients relative to SD controls in the neutral and stress condition shown on the right (significant contrast area shown in red/yellow, p<.01), but not in the alcohol cue condition.
For (d), the medial prefrontal cortex (mPFC) region, group by condition significant effect is shown on the left in red/yellow at [i] p<.005 and [ii] p<.01 thresholds. Simple effect contrasts to explain this significant interaction effect indicated that it was a result of lower activity in mPFC in AD patients relative to SD controls in the stress and alcohol cue conditions shown in pink on the right (p<.01) and not due to a activity change in the neutral condition.

Social interaction was measured 5–6 hrs after ETOH removal. P < 0.01 compared to Control Diet (CD) and the other ETOH exposed groups. Modified from Overstreet et al. (2002) by permission of publisher.

Animals were given three cycles of 5 days of 7% alcohol diet (ED-CY3) followed by an additional 5 days of alcohol diet 8 or 16 days after the initial repeated alcohol exposure (ED-CY3-R-8D & ED-CY3-R-16 D). CD rats received only control diet. The CY1 is withdrawal from a single cycle of alcohol diet to animals that previously received CD. Social interaction as a measure of anxiety-like behavior was measured 5–6 hrs after withdrawal from the re-exposure to alcohol diet. * P < 0.01 compared to CD and CY1. Modified from Overstreet et al. (2002) by permission of publisher.

Dorsal Raphe data are at the top and amygdala data are presented in the bottom illustration. The 5-HT_2C inverse agonist (SB243213; SB), the 5-HT1A agonist (buspirone), and the BZD antagonist (Flumazenil; Flu) were administered into the sites during withdrawal from the initial two cycles. Social interaction as a measure of anxiety-like behavior was measured 5–6 hrs after withdrawal from the final cycle of alcohol. * P < 0.01 compared to control diet exposure. Modified from Overstreet et al (2006) and Knapp et al. (2007) by permission of the publishers.

The (Control Diet) group received only diet. The (No Stress-1 cycle ET) group received no stress and and only 1 cycle ET. The (Control Diet-3 Stress) group received 3 separate 60 min stresses at weekly intervals. The (2 Stresses—1 cycle ET) group was exposed to two 60 min restraint stresses at a weekly interval followed by exposure to 1 cycle of 5 days of 4.5% alcohol diet. Social interaction as a measure of anxiety was measured 5–6 hrs after removal of alcohol diet or the final stress. * P < 0.01 compared to all other groups. From Breese et al. (2004) by permission of publisher.

The CRF1R antagonist was CRA1000, the BZD antagonist was flumazenil (FLU) and the 5-HT1A agonist was buspirone. For each of the drug groups the drug was administered prior to each stress given prior to 5 days of 4.5% ethanol diet (ETOH) Social interaction as a measure of anxiety-like behavior was measured 5–6 hrs after ETOH Diet removal. * P < 0.01 compared to all other groups. From Breese et al. (2004) by permission of publisher.

The CRF-1 receptor antagonist SSR125543 (SSR; 10 μg/0.5 μl) was microinjected into the CeA, DRN, or the dorsal BNST (BNST) 15 min prior to the 2 weekly 60-min restraint stresses applied before exposure to 5 days of 4.5% alcohol diet (ED). For the CD-Vehicle and ED-Vehicle groups, vehicle was administered into each of the brain sites (N = 3–6 for each site) and these data combined. Social interaction, as a measure of anxiety-like behavior, was determined 5–6 hrs after alcohol removal. The CD-Vehicle group was not significantly different from the ED-vehicle group. *P < 0.001 when compared to the CD-Vehicle, ED-Vehicle, and CD Stress groups as well as the groups that received the CRF1R antagonist (SSR) into the three brain sites prior to the repeated stresses. From Huang et al. (2009) by permission of the publisher.

Among 10 distinct areas altered (Gilman and Hommer, 2008), the brain regions exhibiting the greatest activation (P< 0.001) in the alcoholic were the right and left insula, the right inferior frontal cortex, the left lingual cortex, and the right medial temporal cortex. Reproduced with permission from Gilman and Hommer, 2008).

In the placebo group of alcoholics, there were robust significant activations to the application of the negative stimuli in the inferior frontal gyrus, insula, and middle temporal gyrus. The NK1R antagonist-treated group (LY686017) had significantly less activation in these areas. Likewise, the placebo control group had little activation in response to positive emotional stimuli. The NK1R antagonist group of controls exhibited activation in the thalamus, caudate (including the ventral putamen), lingual gyrus and several temporal areas (see George et al., 2008 for details). Modified from George et al. (2008) by permission of the publisher.