Reward value and internal state differentially drive impulsivity and motivation.

Goal-directed behavior is influenced by both reward value as well as internal state. A large body of research has focused on how reward value and internal drives such as hunger influence motivation in rodent models, however less work has focused on how these factors may differentially affect impulsivity. In these studies, we tested the effect of internal drive versus reward value on different facets of reward-related behavior including impulsive action, impulsive choice and, motivation. We varied reward value by changing the concentration of sucrose in the reward outcome, and varied internal drive by manipulating thirst through water restriction. Consistent with the literature we found that both internal state and reward value influenced motivation. However, we found that in high effort paradigms, only internal state influenced motivation with minimal effects of reward value. Interestingly, we found that internal state and reward value differentially influence different subtypes of impulsivity. Internal state, and to a lesser extent, reward value, influenced impulsive action as measured by premature responding. On the other hand, there were minimal effects of either reward value or homeostatic state on impulsive choice as measured by delay discounting. Overall, these studies begin to address how internal state and reward value differentially drive impulsive behavior. Understanding how these factors influence impulsivity is important for developing behavioral interventions and treatment targets for patients with dysregulated motivated or impulsive behavior.

Encoding of conditioned inhibitors of fear in the infralimbic cortex.

Cues in the environment signaling the absence of threat, i.e. safety, can influence both fear and reward-seeking behaviors. Heightened and maladaptive fear is associated with reduced activity in the medial prefrontal cortex. We have previously shown in male rats that the infralimbic (IL) prefrontal cortex is necessary for suppressing fear during a safety cue. The objective of the present study was to determine if there was safety cue-specific neural activity within the IL using a Pavlovian conditioning paradigm, where a fear cue was paired with shock, a safety cue was paired with no shock, and a reward cue was paired with sucrose. To investigate how safety cues can suppress fear, the fear and safety cues were presented together as a compound fear + safety cue. Single-unit activity showed a large proportion of neurons with excitatory responses to the fear + safety cue specifically, a separate group of neurons with excitatory responses to both the reward and fear + safety cues, and bidirectional neurons with excitation to the fear + safety cue and inhibition to the fear cue. Neural activity was also found to be negatively correlated with freezing during the fear + safety cue. Together, these data implicate the IL in encoding specific aspects of conditioned inhibitors when fear is being actively suppressed.

Differential effects of prior stress on conditioned inhibition of fear and fear extinction.

Resistant and generalized fear are hallmark symptoms of Post-Traumatic Stress Disorder (PTSD). Given PTSD is highly comorbid with addiction disorders indicates a maladaptive interaction between fear and reward circuits. To investigate learning processes underlying fear, reward and safety, we trained male rats to discriminate among a fear cue paired with footshock, a reward cue paired with sucrose and an explicit safety cue co-occurring with the fear cue in which no footshocks were delivered. In an attempt to emulate aspects of PTSD, we pre-exposed male rats to a stressor (15 unsignaled footshocks) before training them to fear, reward and safety cues, and subsequent fear and reward extinction. Prior stress did not produce any significant impairments on conditioned inhibition to a safety cue compared to non-stressed controls. However, in subsequent fear extinction, prior stress profoundly impaired fear reduction to an extinguished fear cue. Prior stress also significantly reduced reward seeking to a reward-associated cue throughout training. Together, our data show that prior stress did not affect conditioned inhibition of fear to the same extent as impairing fear extinction. These results have interesting implications on how safety circuits are organized and impacted by stress, leading to possibly new avenues of research on mechanisms of stress disorders, such as PTSD.

Sex differences in fear regulation and reward-seeking behaviors in a fear-safety-reward discrimination task.

Reward availability and the potential for danger or safety potently regulate emotion. Despite women being more likely than men to develop emotion dysregulation disorders, there are comparatively few studies investigating fear, safety and reward regulation in females. Here, we show that female Long Evans rats did not suppress conditioned freezing in the presence of a safety cue, nor did they extinguish their freezing response, whereas males did both. Females were also more reward responsive during the reward cue until the first footshock exposure, at which point there were no sex differences in reward seeking to the reward cue. Darting analyses suggest females were able to regulate this behavior in response to the safety cue, suggesting they were able to discriminate between fear and safety cues but did not demonstrate this with conditioned suppression of freezing behavior. However, levels of darting in this study were too low to make any definitive conclusions. In summary, females showed a significantly different behavioral profile than males in a task that tested the ability to discriminate among fear, safety and reward cues. This paradigm offers a great opportunity to test for mechanisms that are generating these behavioral sex differences in learned safety and reward seeking.

Altering D1 receptor activity in the basolateral amygdala impairs fear suppression during a safety cue.

Accurate discrimination among cues signifying reward, danger or safety initiates the proper emotional response in order to guide behavior. Appropriate conditioned inhibition of fear in the presence of a safety cue would allow an organism to engage in reward seeking behaviors. There is currently little known about the mechanisms of reward, fear and safety cue discrimination and how a safety cue can inhibit fear and release reward seeking from inhibition. Here we assess reward, fear and safety cue learning together using a behavioral paradigm that has identified neurons that discriminate among these cues in the basolateral amygdala (BLA) (Sangha, Chadick, & Janak, 2013). Dopamine signaling in the BLA has been implicated in discriminatory reward learning, learned fear responses and fear extinction. We tested the hypothesis that D1 receptor activity will influence reward-fear-safety cue discrimination by using the D1 receptor agonist, SKF-3839, and antagonist, SCH-23390, either systemically or within the BLA during discrimination learning in male Long Evans rats. We show that both the agonist and antagonist interfered with fear suppression in the presence of the safety cue, when administered systemically or when infused directly into the BLA. This indicates that altering D1 receptor activity in the basolateral amygdala impairs fear suppression during a safety cue. Neither the agonist or antagonist had a consistent negative impact on discriminatory reward seeking when infused into the BLA. However, systemic administration of the D1 receptor agonist did reduce reward seeking behavior during a task that included fear and safety cues. We did not observe a negative impact on reward seeking during systemic administration of a D1 receptor agonist in a task that only included reward cue + sucrose and nonreward cue + no sucrose pairings. This indicates the impairments we saw with the systemically applied agonist in the safety-fear-reward cue discrimination task were more likely due to effects on fear and/or motivation rather than on cue discrimination. Together, our data indicate that altered dopamine D1 receptor activity in the BLA may be a potential mechanism that leads to the impairment in fear suppression to the safety signal seen with PTSD patients.

Memory consolidation within the central amygdala is not necessary for modulation of cerebellar learning.

Amygdala lesions impair, but do not prevent, acquisition of cerebellum-dependent eyeblink conditioning suggesting that the amygdala modulates cerebellar learning. Two-factor theories of eyeblink conditioning posit that a fast-developing memory within the amygdala facilitates slower-developing memory within the cerebellum. The current study tested this hypothesis by impairing memory consolidation within the amygdala with inhibition of protein synthesis, transcription, and NMDA receptors in rats. Rats given infusions of anisomycin or DRB into the central amygdala (CeA) immediately after each eyeblink conditioning session were severely impaired in contextual and cued fear conditioning, but were completely unimpaired in eyeblink conditioning. Rats given the NMDA antagonist ifenprodil into the CeA before each eyeblink conditioning session also showed impaired fear conditioning, but no deficit in eyeblink conditioning. The results indicate that memory formation within the CeA is not necessary for its modulation of cerebellar learning mechanisms. The CeA may modulate cerebellar learning and retention through an attentional mechanism that develops within the training sessions.

Amygdala inactivation impairs eyeblink conditioning in developing rats.

The amygdala facilitates acquisition of eyeblink conditioning in adult animals by enhancing conditioned stimulus (CS) inputs to the cerebellum and the unconditioned response circuitry. Ontogenetic changes in amygdala modulation of eyeblink conditioning have not been investigated directly. We examined the effects of amygdala inactivation on the ontogeny of eyeblink conditioning and conditioned freezing in rat pups. Rat pups received bilateral infusions of saline or bupivacaine into the central nucleus of the amygdala before each of the first five training sessions, which consisted of paired CS-US trials on postnatal days (P) 17-19, P21-23, or P24-26. The final session consisted of CS-alone test trials to assess the effect of amygdala inactivation during training on conditioned freezing. Amygdala inactivation impaired acquisition of eyeblink conditioning in all of the age groups and impaired freezing to the context during the extinction test. The results indicate that the amygdala modulates cerebellar learning as soon as it begins to emerge ontogenetically.

Categorization of photographic images by rats using shape-based image dimensions.

Strong interest exists in developing a rodent model of visual cognition to conduct research into the neural mechanisms of visual categorization. Yet, doubt remains as to whether rats perform visual categorization tasks as do humans and nonhuman primates. Here, we trained eight rats on two visual categorization tasks using photographs of eight objects from each of four basic-level categories: chairs, flowers, cars, and humans. In Experiment 1, rats learned to categorize chairs versus flowers; in Experiment 2, rats learned to categorize cars versus humans. After rats learned each discrimination, we tested them with eight novel pictures from each of the categories. The rats performed at reliably above-chance levels during these generalization tests. To determine which dimension(s) of the stimuli controlled the rats' behavior, we conducted regression analyses using several image dimensions. The chair versus flower discrimination was mainly controlled by the convexity of the stimuli, whereas the car versus human discrimination was mainly controlled by the aspect ratio of the stimuli. These results demonstrate that rats can categorize complex visual objects using shape-based properties of photographs.

Developmental changes in medial auditory thalamic contributions to associative motor learning.

Eyeblink conditioning (EBC) was used in the current study to examine the mechanisms underlying the ontogeny of associative motor learning in rats. Eyeblink conditioning emerges ontogenetically between postnatal day 17 (P17) and P24 in rats. Previous studies used electrical stimulation to show that the ontogeny of EBC is influenced by developmental changes in input from the medial auditory thalamus to the pontine nuclei, which in turn affects input to the cerebellum. The current study used tetrode recordings to examine the ontogeny of medial auditory thalamic sensory responses to the conditioned stimulus (CS) and learning-related activity during EBC. Rat pups were implanted with multiple tetrodes in the medial nucleus of the medial geniculate (MGm) and suprageniculate (SG) and trained on delay EBC on P17-P19, P24-P26, or P31-P33 while recording spike activity. Developmental changes in MGm and SG sensory-related activity were found during a pretraining session with unpaired presentations of the auditory CS and periorbital stimulation unconditioned stimulus (US). Substantial developmental changes were observed in learning-related activity in the MGm and SG during CS-US paired training. The ontogenetic changes in learning-related activity may be related to developmental changes in input to the medial auditory thalamus from the amygdala and cerebellum. The findings suggest that the ontogeny of associative motor learning involves developmental changes in sensory input to the thalamus, amygdala input to the thalamus, thalamic input to the pontine nuclei, and cerebellar feedback to the thalamus.