Pavlovian or associative sensitization and its biological significance.

Pavlovian conditioning is typically distinguished from sensitization but a Pavlovian conditional stimulus (CS) also results in sensitization. A Pavlovian CS can sensitize responding to a probe stimulus that is related to the unconditional stimulus (US) or to the US itself. Pavlovian sensitization has been studied in the defensive, sexual, and feeding systems. In Pavlovian sensitization, the focus is not on a conditional response (CR) directly elicited by the CS but on the response mode that is activated by the CS. Activation of a response mode increases the probability of particular responses and also increases reactivity to various stimuli. Pavlovian sensitization reflects this increased stimulus reactivity. Pavlovian sensitization helps uncover successful learning in situations where a conventional CR does not occur. Pavlovian sensitization also encourages broadening our conceptions of Pavlovian conditioning to include changes in afferent processes. Implications for biological fitness and for basic and translational research are discussed.

Adolescent nicotine exposure promotes adulthood opioid consumption that persists despite adverse consequences and increases the density of insular perineuronal nets.

Adolescence marks a sensitive period for neurodevelopment wherein exposure to drugs of abuse may disrupt maturation and induce persistent changes in neurophysiology which may exacerbate the risk for developing substance use disorders in adulthood. Adolescent nicotine exposure (ANE) enhances motivation to obtain drugs of abuse, particularly opioids, and increases vulnerability for the development of opioid use disorder (OUD). Here, we characterized ANE effects on learning about the adverse consequences of opioid consumption in adulthood in the absence of further nicotine administration. First, we show that ANE engenders punishment resistant fentanyl self-administration in a heterogenous seeking-taking chain schedule of reinforcement at least at the tested dose of fentanyl (0.75 µg/kg). We found that ANE rats consumed significantly more fentanyl and contingent foot shock punishment was less efficacious in limiting fentanyl seeking in ANE rats, relative to nicotine-naïve controls. Next, we demonstrated that ANE limits learning about the deleterious consequences of acute opioid intoxication in adulthood. In a combined conditioned taste avoidance and place preference paradigm we found that ANE resulted in significant reductions in the strength of morphine-induced CTA, and a simultaneous enhancement of CPP at a higher dose that was less capable of driving reinforcement in naïve controls. Finally, we examined the expression of perineuronal nets (PNNs) within insular cortex (IC) and found ANE rats to have increased density of PNNs across the anterior IC and significantly more parvalbumin-labeled IC cells relative to naïve controls. Together, these data lay the framework for a mechanistic explanation of the extreme comorbidity between nicotine use and development of OUDs.

Runway extinction in terrestrial toads (Rhinella arenarum): Instrumental or Pavlovian?

Instrumental appetitive extinction involves the reduction of a previously reinforced response when its occurrence is no longer rewarded. Two experiments with terrestrial toads (Rhinella arenarum) tested whether the occurrence of a nonreinforced response is necessary for response extinction by varying the time of exposure to nonrewarded goal-box stimuli across groups. In Experiment 1, toads that received the same acquisition training (15 sessions, 1 session/day, 300 s of access to water in the goal box) were randomly assigned to two groups. In Group 600 (n=12), animals spent 600 s in the goal box in 8 daily extinction sessions (water present but inaccessible). In Group 0 (n=11), toads performed the runway response (i.e., walking from the start to the goal box) but were removed as soon as they entered the goal box, thus having minimal exposure to nonrewarded goal-box stimuli. The runway response was weakened in Group 600 across extinction trials, but exhibited little change in Group 0. In Experiment 2, toads were randomly assigned to two groups after the same acquisition training. Group 0 (n=7) was treated the same as Group 0 in the previous experiment. In Group RI (retention interval, n=7), toads remained in their home cage for 13 days. Finally, all animals received 4 extinction sessions with 300 s in the empty goal box. There was little behavioral change in Group 0 during the 13 sessions with minimal exposure to the goal box. In extinction, both groups reduced their runway response at similar rates. Although the procedures were instrumental, extinction of the runway response in toads can be accounted for in terms of a Pavlovian approach response to stimuli paired with reward and nonreward in the goal box.

Conceptual-level disgust conditioning in contamination-based obsessive-compulsive disorder.

Pavlovian fear conditioning and extinction represent learning mechanisms underlying exposure-based interventions. While increasing evidence indicates a pivotal role of disgust in the development of contamination-based obsessive-compulsive disorder (C-OCD), dysregulations in conditioned disgust acquisition and maintenance, in particular driven by higher-order conceptual processes, have not been examined. Here, we address this gap by exposing individuals with high (HCC, n = 41) or low (LCC, n = 41) contamination concern to a conceptual-level disgust conditioning and extinction paradigm. Conditioned stimuli (CS+) were images from one conceptual category partially reinforced by unconditioned disgust-eliciting stimuli (US), while images from another category served as non-reinforced conditioned stimuli (CS-). Skin conductance responses (SCRs), US expectancy and CS valence ratings served as primary outcomes to quantify conditioned disgust responses. Relative to LCC, HCC individuals exhibited increased US expectancy and CS+ disgust experience, but comparable SCR levels following disgust acquisition. Despite a decrease in conditioned responses from the acquisition phase to the extinction phase, both groups did not fully extinguish the learned disgust. Importantly, the extinction resilience of acquired disgust was more pronounced in HCC individuals. Together, our findings suggest that individuals with high self-reported contamination concern exhibit increased disgust acquisition and resistance to extinction. The findings provide preliminary evidence on how dysregulated disgust learning mechanism across semantically related concepts may contribute to C-OCD.

Behaviorally conditioned effects of psychoactive drugs in experimental animals: What we have learned from nearly a century of research and what remains to be learned.

Continuous treatment with drugs is a crucial requirement for managing various clinical conditions, including chronic pain and neuropsychiatric disorders such as depression or schizophrenia. Associative learning processes, i.e. Pavlovian conditioning, can play an important role for the effects of drugs and could open new avenues for optimizing patient treatment. In this narrative literature review, we summarize available data in experimental animals regarding the behaviorally conditioned effects of psychostimulants such as d-amphetamine and cocaine, the dopamine receptor agonist apomorphine, the dopamine receptor antagonist haloperidol, morphine and antidepressant drugs. In each section, the drug under discussion is briefly introduced, followed by a detailed examination of conditioning features, including doses and dosing regimens, characteristics of the conditioning process such as test environments or specific conditioned stimuli, testing and conditioned response characteristics, possible extinction or reconditioning or reversal training, neural mechanisms, and finally, the potential clinical relevance of the research area related to the drug. We focus on key outcomes, delve into methodical issues, identify gaps in current knowledge, and suggest future research directions.

Stimulation of VTA dopamine inputs to LH upregulates orexin neuronal activity in a DRD2-dependent manner.

Dopamine and orexins (hypocretins) play important roles in regulating reward-seeking behaviors. It is known that hypothalamic orexinergic neurons project to dopamine neurons in the ventral tegmental area (VTA), where they can stimulate dopaminergic neuronal activity. Although there are reciprocal connections between dopaminergic and orexinergic systems, whether and how dopamine regulates the activity of orexin neurons is currently not known. Here we implemented an opto-Pavlovian task in which mice learn to associate a sensory cue with optogenetic dopamine neuron stimulation to investigate the relationship between dopamine release and orexin neuron activity in the lateral hypothalamus (LH). We found that dopamine release can be evoked in LH upon optogenetic stimulation of VTA dopamine neurons and is also naturally evoked by cue presentation after opto-Pavlovian learning. Furthermore, orexin neuron activity could also be upregulated by local stimulation of dopaminergic terminals in the LH in a way that is partially dependent on dopamine D2 receptors (DRD2). Our results reveal previously unknown orexinergic coding of reward expectation and unveil an orexin-regulatory axis mediated by local dopamine inputs in the LH.

Palatability attributed to alcohol and alcohol-paired flavors.

The orosensory features of alcoholic drinks are potent relapse triggers because they acquire incentive properties during consumption, including enhanced palatability. Whether mice similarly perceive alcoholic drinks to be more palatable after repeated consumption is complicated by reports showing that alcohol elicits aversive taste reactivity responses and conditions flavor avoidance. Here, by analyzing the microstructure of alcohol consumption, we report a gradual increase in lick bout duration relative to water that is partially maintained by an alcohol-paired flavor in extinction. We interpret lick bout duration to reflect an increase in the palatability alcohol and an alcohol-paired flavor. This finding demonstrates that bout duration is amenable to Pavlovian conditioning and highlights the importance of considering the microstructure of alcohol consumption in preclinical models of alcohol misuse.

Threat Expectation Does Not Improve Perceptual Discrimination despite Causing Heightened Priority Processing in the Frontoparietal Network.

Threat cues have been widely shown to elicit increased sensory and attentional neural processing. However, whether this enhanced recruitment leads to measurable behavioral improvements in perception is still in question. Here, we adjudicate between two opposing theories: that threat cues do or do not enhance perceptual sensitivity. We created threat stimuli by pairing one direction of motion in a random dot kinematogram with an aversive sound. While in the MRI scanner, 46 subjects (both men and women) completed a cued (threat/safe/neutral) perceptual decision-making task where they indicated the perceived motion direction of each moving dot stimulus. We found strong evidence that threat cues did not increase perceptual sensitivity compared with safe and neutral cues. This lack of improvement in perceptual decision-making ability occurred despite the threat cue resulting in widespread increases in frontoparietal BOLD activity, as well as increased connectivity between the right insula and the frontoparietal network. These results call into question the intuitive claim that expectation automatically enhances our perception of threat and highlight the role of the frontoparietal network in prioritizing the processing of threat-related environmental cues.

Nucleus accumbens neurons dynamically respond to appetitive and aversive associative learning.

To survive, individuals must learn to associate cues in the environment with emotionally relevant outcomes. This association is partially mediated by the nucleus accumbens (NAc), a key brain region of the reward circuit that is mainly composed by GABAergic medium spiny neurons (MSNs), that express either dopamine receptor D1 or D2. Recent studies showed that both populations can drive reward and aversion, however, the activity of these neurons during appetitive and aversive Pavlovian conditioning remains to be determined. Here, we investigated the relevance of D1- and D2-neurons in associative learning, by measuring calcium transients with fiber photometry during appetitive and aversive Pavlovian tasks in mice. Sucrose was used as a positive valence unconditioned stimulus (US) and foot shock was used as a negative valence US. We show that during appetitive Pavlovian conditioning, D1- and D2-neurons exhibit a general increase in activity in response to the conditioned stimuli (CS). Interestingly, D1- and D2-neurons present distinct changes in activity after sucrose consumption that dynamically evolve throughout learning. During the aversive Pavlovian conditioning, D1- and D2-neurons present an increase in the activity in response to the CS and to the US (shock). Our data support a model in which D1- and D2-neurons are concurrently activated during appetitive and aversive conditioning.

Attenuated incubation of ethanol-induced conditioned taste aversion in a model of dependence.

RATIONALE: Preclinical studies report attenuated ethanol-induced conditioned taste aversion (CTA) following chronic ethanol exposure, suggesting that tolerance develops to the aversive properties of ethanol. However, these studies are confounded by pre-exposure to the unconditioned stimulus (US; ethanol), which is well known to hinder conditioning. OBJECTIVES: This study was designed to determine whether chronic ethanol exposure produces tolerance to the aversive properties of ethanol in the absence of a US pre-exposure confound. METHODS: CTA was performed in adult male and female Long-Evans rats by pairing 0.1% ingested saccharin with an intraperitoneal injection of ethanol (1.5 or 2.0 g/kg) or saline. Rats were then rendered ethanol dependent using chronic intermittent ethanol (CIE) vapor exposure. Controls were exposed to room air (AIR). The effect of chronic ethanol on CTA expression and reconditioning were examined following vapor exposure. RESULTS: Prior to vapor exposure, both sexes developed CTA to a comparable degree with 2.0 g/kg producing greater CTA than 1.5 g/kg ethanol. Following vapor exposure, AIR controls exhibited an increase in CTA magnitude compared to pre-vapor levels. This effect was largely absent in CIE-exposed rats. Re-conditioning after vapor exposure facilitated increased CTA magnitude to a similar degree in AIR- and CIE-exposed males. In contrast, CTA magnitude was unchanged by re-conditioning in females. CONCLUSIONS: These data suggest that chronic ethanol does not facilitate tolerance to the aversive properties of ethanol but rather attenuates incubation of ethanol-induced CTA. Loss of CTA incubation suggests that CIE exposure disrupts circuits encoding aversion.

Encoding of Predictive Associations in Human Prefrontal and Medial Temporal Neurons During Pavlovian Appetitive Conditioning.

Pavlovian conditioning is thought to involve the formation of learned associations between stimuli and values, and between stimuli and specific features of outcomes. Here, we leveraged human single neuron recordings in ventromedial prefrontal, dorsomedial frontal, hippocampus, and amygdala while patients of both sexes performed an appetitive Pavlovian conditioning task probing both stimulus-value and stimulus-stimulus associations. Ventromedial prefrontal cortex encoded predictive value along with the amygdala, and also encoded predictions about the identity of stimuli that would subsequently be presented, suggesting a role for neurons in this region in encoding predictive information beyond value. Unsigned error signals were found in dorsomedial frontal areas and hippocampus, potentially supporting learning of non-value related outcome features. Our findings implicate distinct human prefrontal and medial temporal neuronal populations in mediating predictive associations which could partially support model-based mechanisms during Pavlovian conditioning.

Lateral hypothalamic GABAergic neurons encode alcohol memories.

In alcohol use disorder, the alcohol memories persist during abstinence, and exposure to stimuli associated with alcohol use can lead to relapse. This highlights the importance of investigating the neural substrates underlying not only relapse but also encoding and expression of alcohol memories. GABAergic neurons in the lateral hypothalamus (LH-GABA) have been shown to be critical for food-cue memories and motivation; however, the extent to which this role extends to alcohol-cue memories and motivations remains unexplored. In this study, we aimed to describe how alcohol-related memories are encoded and expressed in LH GABAergic neurons. Our first step was to monitor LH-GABA calcium transients during acquisition, extinction, and reinstatement of an alcohol-cue memory using fiber photometry. We trained the rats on a Pavlovian conditioning task, where one conditioned stimulus (CS+) predicted alcohol (20% EtOH) and another conditioned stimulus (CS-) had no outcome. We then extinguished this association through non-reinforced presentations of the CS+ and CS- and finally, in two different groups, we measured relapse under non-primed and alcohol-primed induced reinstatement. Our results show that initially both cues caused increased LH-GABA activity, and after learning only the alcohol cue increased LH-GABA activity. After extinction, this activity decreases, and we found no differences in LH-GABA activity during reinstatement in either group. Next, we inhibited LH-GABA neurons with optogenetics to show that activity of these neurons is necessary for the formation of an alcohol-cue association. These findings suggest that LH-GABA might be involved in attentional processes modulated by learning.

Augmenting glutamatergic, but not dopaminergic, activity in the nucleus accumbens shell disrupts responding to a discrete alcohol cue in an alcohol context.

Discrete alcohol cues and contexts are relapse triggers for people with alcohol use disorder exerting particularly powerful control over behaviour when they co-occur. Here, we investigated the neural substrates subserving the capacity for alcohol-associated contexts to elevate responding to an alcohol-predictive conditioned stimulus (CS). Specifically, rats were trained in a distinct 'alcohol context' to respond by entering a fluid port during a discrete auditory CS that predicted the delivery of alcohol and were familiarized with a 'neutral context' wherein alcohol was never available. When conditioned CS responding was tested by presenting the CS without alcohol, we found that augmenting glutamatergic activity in the nucleus accumbens (NAc) shell by microinfusing α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) reduced responding to an alcohol CS in an alcohol, but not neutral, context. Further, AMPA microinfusion robustly affected behaviour, attenuating the number, duration and latency of CS responses selectively in the alcohol context. Although dopaminergic inputs to the NAc shell were previously shown to be necessary for CS responding in an alcohol context, here, chemogenetic excitation of ventral tegmental area (VTA) dopamine neurons and their inputs to the NAc shell did not affect CS responding. Critically, chemogenetic excitation of VTA dopamine neurons affected feeding behaviour and elevated c-fos immunoreactivity in the VTA and NAc shell, validating the chemogenetic approach. These findings enrich our understanding of the substrates underlying Pavlovian responding for alcohol and reveal that the capacity for contexts to modulate responding to discrete alcohol cues is delicately underpinned by the NAc shell.

Conditioning to true content and artificial intelligence in psychophysiological intention recognition.

OBJECTIVE: The objective is to introduce a novel method for classical conditioning to true content (CtTC), and for the first time, apply this approach in the concealed information test (CIT) to effectively discern intentions. During CtTC, participants are trained to exhibit electrodermal responses whenever they recognize true content on a screen. Additionally, the objective is to evaluate a novel CIT-dataset preprocessing algorithm, employed to enhance machine learning (ML) classification performance. METHODS: A total of 84 participants were evenly divided into four groups. Two groups of participants devised plans for stealing money from a supermarket, while the other two groups did not engage in any planning. One planning group and one non-planning group underwent CIT examination, while the remaining groups were subjected to CtTC. RESULTS: The CIT accuracy initially stood at 52 % and increased to 71 % after Z-score and ML classification (McNemar test, p < 0.05). Conversely, the CtTC accuracy was 76 % and significantly improved to 93 % following Z-score and 95 % following ML classification (McNemar test, p < 0.05). In the best-performing classifiers, CtTC exhibited significantly superior metrics for guilty/innocent classification compared to CIT (Fisher's exact test, p < 0.05, power 1 - ß > 0.90). In the CtTC group, reactivity and sensitivity significantly increased, indicated by higher EDR amplitudes (p < 0.05, two-tailed t-test, power 1 - ß = 0.89) and the number of EDRs (p < 0.05, Fisher's exact test, power 1 - ß = 0.90). There was no statistically significant difference between the Z-score and ML classification. CONCLUSIONS: In the assessment of intentions, CtTC enhances both the sensitivity and accuracy of the CIT.

The effects of extinction and an explicitly unpaired treatment on the reinforcing properties of a Pavlovian conditioned stimulus.

This series of experiments examined the effects of extinction and an explicitly unpaired treatment on the ability of a conditioned stimulus (CS) to function as a reinforcer. Rats were trained to lever press for food, exposed to pairings of a noise CS and food, and, finally, tested for their willingness to lever press for the CS in the absence of the food. Experiment 1 provided a demonstration of conditioned reinforcement (using controls that were only exposed to unpaired presentations of the CS and food) and showed that it was equivalent after one or four sessions of CS-food pairings. Experiments 2 and 3 showed that, after one session of CS-food pairings, repeated presentations of the CS alone reduced its reinforcing properties; but after four sessions of CS-food pairings, repeated presentations of the CS alone had no effect on these properties. Experiment 4 showed that, after four sessions of CS-food pairings, explicitly unpaired presentations of the CS and food completely undermined conditioned reinforcement. Finally, Experiment 5 provided within-experiment evidence that, after four sessions of CS-food pairings, the reinforcing properties of the CS were disrupted by explicitly unpaired presentations of the CS and food but spared by repeated presentations of the CS alone. Together, these findings indicate that the effectiveness of extinction in undermining the reinforcing properties of a CS depends on its level of conditioning; and that, where extinction fails to disrupt these properties, they are successfully undermined by an explicitly unpaired treatment. They are discussed with respect to findings in the literature on Pavlovian-to-instrumental transfer; and the Rescorla-Wagner model, which anticipates that an explicitly unpaired treatment will be more effective than extinction in reversing the effects of conditioning.

Reversal of Conditioned Food Aversion Using a Cognitive Intervention: A Sham-Controlled, Randomized, Parallel Study.

BACKGROUND: Aversive conditioning weakens the gratifying value of a comfort meal. The aim was to determine the effect of a cognitive intervention to reverse aversive conditioning and restore hedonic postprandial response. METHODS: This was a randomized, sham-controlled, single-blind, parallel study that was conducted on 12 healthy women (n = 6 in each group). The reward value of a comfort meal was measured on different days: at initial exposure, after aversive conditioning (administration of the same meal with a masked fat overload on the previous day) and after a cognitive intervention (disclosing the aversive conditioning paradigm in the test group vs. no explanation in the control group). The primary outcome, digestive wellbeing, was determined using graded scales at regular intervals before and after ingestion. RESULTS: At initial exposure, the comfort meal produced a rewarding experience that was impaired using aversive conditioning; upon re-exposure to the original meal, the cognitive intervention increased meal wanting and liking; improved digestive wellbeing and mood; tended to reduce postprandial satiety, bloating/fullness; and abolished discomfort/pain, thereby restoring the hedonic value of the comfort meal. By contrast, sham intervention had no effects, and the postprandial sensations remained like the responses to the offending meal. CONCLUSION: In this proof-of-concept study, we demonstrate that in healthy women, a mild, short-term acquired aversion to a comfort meal can be reversed using a cognitive intervention. CLINICALTRIALS: gov ID: NCT05897411.

Propensity for risky choices despite lower cue reactivity in adolescent rats.

Adolescence is a time of heightened risk-taking across species. Salient audiovisual cues associated with rewards are a common feature of gambling environments and have been connected to increased risky decision-making. We have previously shown that, in adult male rats, sign tracking - a behavioral measure of cue reactivity - predicts an individual's propensity for suboptimal risky choices in a rodent gambling task (rGT) with win-paired cues. However, adolescents perform less sign tracking than adult animals, suggesting that they are less cue-reactive than adults in some circumstances. Therefore, we investigated the performance of adolescent male rats on the rGT with win cues and examined its relationship with their sign-tracking behavior. We found that adolescents make more risky choices and fewer optimal choices on the rGT compared with adults, evidence of the validity of the rGT as a model of adolescent gambling behavior. We also confirmed that adolescents perform less sign tracking than adults, and we found that, unlike in adults, adolescents' sign tracking was unrelated to their risk-taking in the rGT. This implies that adolescent risk-taking is less likely than that of adults to be driven by reward-related cues. Finally, we found that adults trained on the rGT as adolescents retained an adolescent-like propensity toward risky choices, suggesting that early exposure to a gambling environment may have a long-lasting impact on risk-taking behavior.

Elevated Amygdala Responses During De Novo Pavlovian Conditioning in Alcohol Use Disorder Are Associated With Pavlovian-to-Instrumental Transfer and Relapse Latency.

Background: Contemporary learning theories of drug addiction ascribe a key role to Pavlovian learning mechanisms in the development, maintenance, and relapse of addiction. In fact, cue-reactivity research has demonstrated the power of alcohol-associated cues to activate the brain's reward system, which has been linked to craving and subsequent relapse. However, whether de novo Pavlovian conditioning is altered in alcohol use disorder (AUD) has rarely been investigated. Methods: To characterize de novo Pavlovian conditioning in AUD, 62 detoxified patients with AUD and 63 matched healthy control participants completed a Pavlovian learning task as part of a Pavlovian-to-instrumental transfer paradigm during a functional magnetic resonance imaging session. Patients were followed up for 12 months to assess drinking behavior and relapse status. Results: While patients and healthy controls did not differ in their ability to explicitly acquire the contingencies between conditioned and unconditioned stimuli, patients with AUD displayed significantly stronger amygdala responses toward Pavlovian cues, an effect primarily driven by stronger blood oxygen level-dependent differentiation during learning from reward compared with punishment. Moreover, in patients compared with controls, differential amygdala responses during conditioning were positively related to the ability of Pavlovian stimuli to influence ongoing instrumental choice behavior measured during a subsequent Pavlovian-to-instrumental transfer test. Finally, patients who relapsed within the 12-month follow-up period showed an inverse association between amygdala activity during conditioning and relapse latency. Conclusions: We provide evidence of altered neural correlates of de novo Pavlovian conditioning in patients with AUD, especially for appetitive stimuli. Thus, heightened processing of Pavlovian cues might constitute a behaviorally relevant mechanism in alcohol addiction.

The next frontier: Moving human fear conditioning research online.

Fear conditioning is a significant area of research that has featured prominently among the topics published in Biological Psychology over the last 50 years. This work has greatly contributed to our understanding of human anxiety and stressor-related disorders. While mainly conducted in the laboratory, recently, there have been initial attempts to conduct fear conditioning experiments online, with around 10 studies published on the subject, primarily in the last two years. These studies have demonstrated the potential of online fear conditioning research, although challenges to ensure that this research meets the same methodological standards as in-person experimentation remain, despite recent progress. We expect that in the coming years new outcome measures will become available online including the measurement of eye-tracking, pupillometry and probe reaction time and that compliance monitoring will be improved. This exciting new approach opens new possibilities for large-scale data collection among hard-to-reach populations and has the potential to transform the future of fear conditioning research.

Prediction error in models of adaptive behavior.

Pavlovian conditioning is evident in every species in which it has been assessed, and there is a consensus about its interpretation across behavioral,1,2 brain,3,4,5,6 and computational analyses7,8,9,10,11: conditioned behavior reflects the formation of a directional associative link from the memory of one stimulus (e.g., a visual stimulus) to another (e.g., food), with learning stopping when there is no error between the prediction generated by the visual stimulus and what happens next (e.g., food). This consensus fails to anticipate the results that we report here. In our experiments with rats, we find that arranging predictive (visual stimulus→food) and nonpredictive (food→visual stimulus) relationships produces marked and sustained changes in conditioned behaviors when the visual stimulus is presented alone. Moreover, the type of relationship affects (1) the distribution of conditioned behaviors related to the properties of both food (called goal-tracking) and the visual stimulus (called sign-tracking) and (2) when in the visual stimulus, these two behaviors are evident. These results represent an impetus for a fundamental shift in how Pavlovian conditioning is interpreted: animals learn about the relationship between two stimuli irrespective of the order in which they are presented, but they exhibit this knowledge in different ways. This interpretation and our new results are captured by a recent model of Pavlovian conditioning,12,13 HeiDI, and both are consistent with the need for animals to represent the fact that the impact of a cause (e.g., the ingestion of nutrients or the bite of a predator) can be felt before or after the cause has been perceived.