Medial prefrontal cortex neuronal activation and synaptic alterations after stress-induced reinstatement of palatable food seeking: a study using c-fos-GFP transgenic female rats

Relapse to maladaptive eating habits during dieting is often provoked by stress and there is evidence for a role of ovarian hormones in stress responses and feeding. We studied the role of these hormones in stress-induced reinstatement of food seeking and medial prefrontal cortex (mPFC) neuronal activation in c-fos-GFP transgenic female rats, which express GFP in strongly activated neurons. Food-restricted ovariectomized or sham-operated c-fos-GFP rats were trained to lever-press for palatable food pellets. Subsequently, lever-pressing was extinguished and reinstatement of food seeking and mPFC neuronal activation was assessed after injections of the pharmacological stressor yohimbine (0.5-2 mg/kg) or pellet priming (1-4 noncontingent pellets). Estrous cycle effects on reinstatement were also assessed in wild-type rats. Yohimbine- and pellet-priming-induced reinstatement was associated with Fos and GFP induction in mPFC; both reinstatement and neuronal activation were minimally affected by ovarian hormones in both c-fos-GFP and wild-type rats. c-fos-GFP transgenic rats were then used to assess glutamatergic synaptic alterations within activated GFP-positive and nonactivated GFP-negative mPFC neurons following yohimbine-induced reinstatement of food seeking. This reinstatement was associated with reduced AMPA receptor/NMDA receptor current ratios and increased paired-pulse facilitation in activated GFP-positive but not GFP-negative neurons. While ovarian hormones do not appear to play a role in stress-induced relapse of food seeking in our rat model, this reinstatement was associated with unique synaptic alterations in strongly activated mPFC neurons. Our paper introduces the c-fos-GFP transgenic rat as a new tool to study unique synaptic changes in activated neurons during behavio

Individual variability in behavioral flexibility predicts sign-tracking tendency

Sign-tracking rats show heightened sensitivity to food- and drug-associated cues, which serve as strong incentives for driving reward seeking. We hypothesized that this enhanced incentive drive is accompanied by an inflexibility when incentive value changes. To examine this we tested rats in Pavlovian outcome devaluation or second-order conditioning prior to the assessment of sign-tracking tendency. To assess behavioral flexibility we trained rats to associate a light with a food outcome. After the food was devalued by pairing with illness, we measured conditioned responding to the light during an outcome devaluation probe test. The level of conditioned responding during outcome devaluation probe test correlated with the rats’ subsequent tracking tendency, with sign-tracking rats failing to suppress conditioned responding to the light after outcome devaluation. To assess Pavlovian incentive learning, we trained rats on first-order (CS+, CS-) and second-order (SOCS+, SOCS-) discriminations. After second-order conditioning, we measured conditioned responding to the second-order cues during a probe test. Second-order conditioning was observed in all rats regardless of tracking tendency. The behavioral inflexibility of sign-trackers has potential relevance for understanding individual variation in vulnerability to drug addiction

Stability of individual differences in sucralose taste preference.

Outbred rats display variable preferences for bittersweet solutions, expressed as preference or avoidance of high concentrations of artificial sweeteners over water. This may reflect individual differences in appetitive/aversive conflict processing that may have predictive validity for disorders of motivation. Here we use a homecage two-bottle choice procedure to examine the test/retest stability and between-tastant consistency in sucralose preference to determine the reliability of bittersweet taste preference. Sucralose is a non-caloric artificial sweetener that is preferred by some rats and avoided by others. We sought to determine whether sucralose preference is consistent with preference of sucrose/quinine solutions that have known sweet and bitter taste qualities, respectively. We give fluid restricted rats 45-minutes homecage access to water and ascending concentrations of sucralose (SUCRA; 0.0025-10mM) or a compound solution of sucrose (116mM) + quinine (0.002-2mM) (SQ). We use a within-subject counterbalanced design (SUCRA or SQ testing) to determine preference of each bittersweet solution relative to water. We observed individual variability in preference for SUCRA and SQ, such that some rats preferred bittersweet solutions over water (preferring) while other rats preferred water over bittersweet solutions (avoiding). Within tastant, this preference remained stable across repeated testing. Between solutions, SUCRA preference scores correlated with SQ scores, suggesting consistent taste conflict processing for both bittersweet solutions. Population level analyses confirmed that preference generalizes across bittersweet solutions, and that rats' preferences for bittersweet solutions relative to water are stable over time. The test/retest and between-tastant reliability of this taste conflict screening procedure support the potential utility of this model for exploring individual variability in appetitive/aversive conflict processes mediating motivated behavior

The reinstatement model of drug relapse: Recent neurobiological findings, emerging research topics, and translational research

Background and rationale: Results from many clinical studies suggest that drug relapse and craving are often provoked by acute exposure to the self-administered drug or related drugs, drug-associated cues or contexts, or certain stressors. During the last two decades, this clinical scenario has been studied in laboratory animals by using the reinstatement model. In this model, reinstatement of drug seeking by drug priming, drug cues or contexts, or certain stressors is assessed following drug self-administration training and subsequent extinction of the drug-reinforced responding. Objective: In this review, we first summarize recent (2009-present) neurobiological findings from studies using the reinstatement model. We then discuss emerging research topics, including the impact of interfering with putative reconsolidation processes on cue- and context-induced reinstatement of drug seeking, and similarities and differences in mechanisms of reinstatement across drug classes. We conclude by discussing results from recent human studies that were inspired by results from rat studies using the reinstatement model. Conclusions: Main conclusions from the studies reviewed highlight: (1) the ventral subiculum and lateral hypothalamus as emerging brain areas important for reinstatement of drug seeking, (2) the existence of differences in brain mechanisms controlling reinstatement of drug seeking across drug classes, (3) the utility of the reinstatement model for assessing the effect of reconsolidation-related manipulations on cue-induced drug seeking, and (4) the encouraging pharmacological concordance between results from rat studies using the reinstatement model and human laboratory studies on cue- and stress-induced drug craving

Effect of cafeteria diet history on cue-, pellet-priming-, and stress-induced reinstatement of food seeking in female rats.

Relapse to unhealthy eating habits is a major problem in human dietary treatment. The individuals most commonly seeking dietary treatment are overweight or obese women, yet the commonly used rat reinstatement model to study relapse to palatable food seeking during dieting primarily uses normal-weight male rats. To increase the clinical relevance of the relapse to palatable food seeking model, here we pre-expose female rats to a calorically-dense cafeteria diet in the home-cage to make them overweight prior to examining the effect of this diet history on cue-, pellet-priming- and footshock-induced reinstatement of food seeking.Post-natal day 32 female Long-Evans rats had seven weeks of home-cage access to either chow only or daily or intermittent cafeteria diet alongside chow. Next, they were trained to self-administer normally preferred 45 mg food pellets accompanied by a tone-light cue. After extinction, all rats were tested for reinstatement induced by discrete cue, pellet-priming, and intermittent footshock under extinction conditions.Access to daily cafeteria diet and to a lesser degree access to intermittent cafeteria diet decreased food pellet self-administration compared to chow-only. Prior history of these cafeteria diets also reduced extinction responding, cue- and pellet-priming-induced reinstatement. In contrast, modest stress-induced reinstatement was only observed in rats with a history of daily cafeteria diet.A history of cafeteria diet does not increase the propensity for cue- and pellet-priming-induced relapse in the rat reinstatement model but does appear to make rats more susceptible to footshock stress-induced reinstatement

Neural Correlates of Variations in Event Processing during Learning in Central Nucleus of Amygdala

SummaryAttention or variations in event processing help drive learning. Lesion studies have implicated the central nucleus of the amygdala (CeA) in this process, particularly when expected rewards are omitted. However, lesion studies cannot specify how information processing in CeA supports such learning. To address these questions, we recorded CeA neurons in rats performing a task in which rewards were delivered or omitted unexpectedly. We found that activity in CeA neurons increased selectively at the time of omission and declined again with learning. Increased firing correlated with CeA-inactivation sensitive measures of attention. Notably CeA neurons did not fire to the cues or in response to unexpected rewards. These results indicate that CeA contributes to learning in response to reward omission due to a specific role in signaling actual omission rather than a more general involvement in signaling expectancies, errors, or reward value

Summary of inactive-lever presses during the training, extinction, and reinstatement testing.

<p>Data are mean±SEM.</p

Effect of home-cage diet condition on food self-administration training.

<p><b>A.</b> Reinforcers (pellets) earned (mean±SEM). <b>B.</b> Time-out responding during food self-administration training for different diet conditions. Inset: difference in time-out between training sessions 1 and 10 for each diet condition. ^# Different between sessions 1 and 10 within diet condition, p<0.05. <b>C.</b> Active lever responding (mean±SEM) under extinction conditions without cue (lever responding had no consequence).*Different between chow-only and daily cafeteria group, p<0.05, n = 8 per group. Data points with heavier outlines indicate days that intermittent group had access to cafeteria items prior to the session.</p

Effect of home-cage diet histories on reinstatement of food seeking.

<p><b>A.</b> Effect of discrete cue presentation on reinstatement. Active lever responding (mean±SEM) under extinction (with cue) conditions following a single non-contingent cue presentation at the start of the session. *Different between the last day of extinction (without cue) and cue-test within subject, p<0.05. ^# Different in overall lever pressing from chow-only group, p = 0.06. <b>B.</b> Effect of pellet priming on reinstatement. Active lever responding (mean±SEM) under extinction conditions (with cue) following 1, 2, and 4 non-contingent pellet deliveries at the beginning of an extinction session. *Different between 0 pellet baseline (last one hour extinction session) and respective pellet test within subject, p<0.05. ^# Different in overall lever pressing from chow-only group, p<0.05. <b>C.</b> Effect of intermittent footshock on reinstatement. Active lever responding (mean±SEM) under extinction conditions (with cue) following 5 min and 10 min of intermittent footshock.</p