Aversive Learning Research Articles

Quantifying population-level neural tuning functions using Ricker wavelets and the Bayesian bootstrap

BackgroundExperience changes visuo-cortical tuning. In humans, re-tuning has been studied during aversive generalization learning, in which the similarity of generalization stimuli (GSs) with a conditioned threat cue (CS+) is used to quantify tuning functions. Previous work utilized pre-defined tuning shapes (generalization and sharpening patterns). This approach may constrain the ways in which re-tuning can be characterized since the tuning patterns may not match the prototypical functions. New methodThe present study proposes a flexible and data-driven method for precisely quantifying changes in tuning based on the Ricker wavelet function and the Bayesian bootstrap. This method was applied to EEG and psychophysics data from an aversive generalization learning paradigm. ResultsThe Ricker wavelet model fitted the steady-state visual event potentials (ssVEP), alpha-band power, and detection accuracy data well. A Morlet wavelet function was used for comparison and fit the data better in some situations, but was more challenging to interpret. The pattern of re-tuning in the EEG data, predicted by the Ricker model, resembled the shapes of the best fitting a-priori patterns. Comparison with existing methodsAlthough the re-tuning shape modeled by the Ricker function resembled the pre-defined shapes, the Ricker approach led to greater Bayes factors and more interpretable results compared to a-priori models. The Ricker approach was more easily fit and led to more interpretable results than a Morlet wavelet model. ConclusionThis work highlights the promise of the current method for capturing the precise nature of visuo-cortical tuning, unconstrained by the implementation of a-priori models.

Running-based food aversion learning in freely-fed and hydrated rats: Daily monitoring of running-induced nausea by measuring kaolin clay ingestion

The main objective of this research was to demonstrate food aversion learning in rats with unrestricted access to food and water, using wheel running as the unconditioned stimulus. Experiment 1 showed that the target-running paired training group consumed a statistically smaller amount of the target food (tteok rice cakes) compared to the target/running unpaired control group, but the decrease in consumption over days in the paired group was not fully supported by a statistical test. Experiment 2a improved the methodology by familiarizing rats with tteok before training, which resulted in both a statistically significant group effect and a statistically significant daily decrease in tteok consumption. Experiment 2b demonstrated that tteok aversion could be reacquired after an extinction phase. These experiments indicate that running-based tteok aversion in non-deprived rats is a valid example of Pavlovian conditioning and suggest that wheel running can cause similar effects in unrestricted rats as observed in food- or water-restricted rats. Additionally, daily measurements of kaolin clay ingestion suggested that wheel running induced nausea in the rats of these experiments.

Classical Conditioning of Adult Drosophila.

Olfactory classical conditioning paradigms have been extensively used since the early 1970s to apply genetic approaches to the study of memory in Drosophila. Over the intervening years, advances in genetics have largely changed the focus of studies from the role of single genes in memory to investigation of memory-relevant neuronal circuits. However, the original behavioral paradigms have remained largely unaltered, besides investigators making a few useful tweaks to the training and testing apparatus and modifications to the operating procedures. In this protocol, we provide the reader with a detailed description of the manufacture and assembly of a typical T-maze apparatus, where populations of adult flies can be trained and their odor memory tested later, by giving them a binary choice between the two trained odors. We describe how variations of the training apparatus permit both aversive (odor-shock) and appetitive (odor-sugar) memories to be studied. In addition, we describe a recent modification of the apparatus and protocol that permits study of multisensory (color and odor) aversive and appetitive learning. Control assays for sensory acuity and locomotion are also included.

Reward and punishment contingency shifting reveals distinct roles for VTA dopamine and GABA neurons in behavioral flexibility.

In dynamic environments where stimuli predicting rewarding or aversive outcomes unexpectedly change, it is critical to flexibly update behavior while preserving recollection of previous associations. Dopamine and GABA neurons in the ventral tegmental area (VTA) are implicated in reward and punishment learning, yet little is known about how each population adapts when the predicted outcome valence changes. We measured VTA dopamine and GABA population activity while male and female rats learned to associate three discrete auditory cues to three distinct outcomes: reward, punishment, or no outcome within the same session. After learning, the reward and punishment cue-outcome contingencies were reversed, and subsequently rereversed. As expected, the dopamine population rapidly adapted to learning and contingency reversals by increasing the response to appetitive stimuli and decreasing the response to aversive stimuli. In contrast, the GABA population increased activity to all sensory events regardless of valence, including the neutral cue. Reversing learned contingencies selectively influenced GABA responses to the reward-predictive cue, prolonging increased activity within and across sessions. The observed valence-specific dissociations in the directionality and temporal progression of VTA dopamine and GABA calcium activity indicates that these populations are independently recruited and serve distinct roles during appetitive and aversive associative learning and contingency reversal.

Aversive sexual learning in the kissing bug Rhodnius prolixus: Modulation of different sexual responses in males and females

Although sexual learning can be a key process in the reproductive success of animals, research focused on the experience-dependent modulation of courtship in insects is scarce. In kissing bugs, the behavioural steps implicated in mating have been exhaustively studied, but not the involvement of learning in them. Our objective was to determine whether the sexual behaviour of Rhodnius prolixus could be modulated by experience. During training, couples were submitted to eight assays, in which they received a vibration (negative reinforcement) when the male attempted to copulate the female. Immediately after, they were separated, not allowing the occurrence of copulation. We found that along training, males’ latency to perform a copulatory attempt increased, male’s copulatory attempts were less frequent, and females’ locomotor activity did not change. These results suggest that males, and not females, learned to avoid the vibration by reducing their intention to copulate. In post-training tests, conditioned males presented with new naïve females reverted to low copulatory attempt latencies, suggesting that the modulation was partner-specific. Besides, conditioned females increased their rejection frequencies to males’ copulatory attempts, suggesting that a second type of learning occurred in females. These results constitute the first evidence of sexual learning in hematophagous insects. Males and females seem to change their selectivity according to their previous sexual experience. We discuss the relevance that this plasticity might have in the fitness of this epidemiologically relevant insect.

Inhibitory mechanisms in the prefrontal-cortex differentially mediate Putamen activity during valence-based learning.

Learning from appetitive and aversive stimuli involves interactions between the prefrontal cortex and subcortical structures. Preclinical and theoretical studies indicate that inhibition is essential in regulating the relevant neural circuitry. Here, we demonstrate that GABA, the main inhibitory neurotransmitter in the central nervous system, differentially affects how the dACC interacts with subcortical structures during appetitive and aversive learning in humans. Participants engaged in tasks involving appetitive and aversive learning, while using functional magnetic resonance spectroscopy (MRS) at 7T to track GABA concentrations in the dACC, alongside whole-brain fMRI scans to assess BOLD activation. During appetitive learning, dACC GABA concentrations were negatively correlated with learning performance and BOLD activity measured from the dACC and the Putamen. These correlations were absent during aversive learning, where dACC GABA concentrations negatively correlated with the connectivity between the dACC and the Putamen. Our results show that inhibition in the dACC mediates appetitive and aversive learning in humans through distinct mechanisms.

Functional connectivity between the nucleus accumbens and amygdala underlies avoidance learning during adolescence: Implications for developmental psychopathology.

Reward and threat processes work together to support adaptive learning during development. Adolescence is associated with increasing approach behavior (e.g., novelty-seeking, risk-taking) but often also coincides with emerging internalizing symptoms, which are characterized by heightened avoidance behavior. Peaking engagement of the nucleus accumbens (NAcc) during adolescence, often studied in reward paradigms, may also relate to threat mechanisms of adolescent psychopathology. 47 typically developing adolescents (9.9-22.9 years) completed an aversive learning task during functional magnetic resonance imaging, wherein visual cues were paired with an aversive sound or no sound. Task blocks involved an escapable aversively reinforced stimulus (CS+r), the same stimulus without reinforcement (CS+nr), or a stimulus that was never reinforced (CS-). Parent-reported internalizing symptoms were measured using Revised Child Anxiety and Depression Scales. Functional connectivity between the NAcc and amygdala differentiated the stimuli, such that connectivity increased for the CS+r (p = .023) but not for the CS+nr and CS-. Adolescents with greater internalizing symptoms demonstrated greater positive functional connectivity for the CS- (p = .041). Adolescents show heightened NAcc-amygdala functional connectivity during escape from threat. Higher anxiety and depression symptoms are associated with elevated NAcc-amygdala connectivity during safety, which may reflect poor safety versus threat discrimination.

A neural mechanism for learning from delayed postingestive feedback.

Animals learn the value of foods based on their postingestive effects and thereby develop aversions to foods that are toxic1-6 and preferences to those that are nutritious7-14. However, it remains unclear how the brain is able to assign credit to flavors experienced during a meal with postingestive feedback signals that can arise after a substantial delay. Here, we reveal an unexpected role for postingestive reactivation of neural flavor representations in this temporal credit assignment process. To begin, we leverage the fact that mice learn to associate novel15-18, but not familiar, flavors with delayed gastric malaise signals to investigate how the brain represents flavors that support aversive postingestive learning. Surveying cellular resolution brainwide activation patterns reveals that a network of amygdala regions is unique in being preferentially activated by novel flavors across every stage of the learning process: the initial meal, delayed malaise, and memory retrieval. By combining high-density recordings in the amygdala with optogenetic stimulation of genetically defined hindbrain malaise cells, we find that postingestive malaise signals potently and specifically reactivate amygdalar novel flavor representations from a recent meal. The degree of malaise-driven reactivation of individual neurons predicts strengthening of flavor responses upon memory retrieval, leading to stabilization of the population-level representation of the recently consumed flavor. In contrast, meals without postingestive consequences degrade neural flavor representations as flavors become familiar and safe. Thus, our findings demonstrate that interoceptive reactivation of amygdalar flavor representations provides a neural mechanism to resolve the temporal credit assignment problem inherent to postingestive learning.

Isolating the koniocellular contribution to aversive learning in human visual cortex

Isolating the koniocellular contribution to aversive learning in human visual cortex

Signal flow in the NMDA receptor-dependent phosphoproteome regulates postsynaptic plasticity for aversive learning.

Structural plasticity of dendritic spines in the nucleus accumbens (NAc) is crucial for learning from aversive experiences. Activation of NMDA receptors (NMDARs) stimulates Ca2+-dependent signaling that leads to changes in the actin cytoskeleton, mediated by the Rho family of GTPases, resulting in postsynaptic remodeling essential for learning. We investigated how phosphorylation events downstream of NMDAR activation drive the changes in synaptic morphology that underlie aversive learning. Large-scale phosphoproteomic analyses of protein kinase targets in mouse striatal/accumbal slices revealed that NMDAR activation resulted in the phosphorylation of 194 proteins, including RhoA regulators such as ARHGEF2 and ARHGAP21. Phosphorylation of ARHGEF2 by the Ca2+-dependent protein kinase CaMKII enhanced its RhoGEF activity, thereby activating RhoA and its downstream effector Rho-associated kinase (ROCK/Rho-kinase). Further phosphoproteomic analysis identified 221 ROCK targets, including the postsynaptic scaffolding protein SHANK3, which is crucial for its interaction with NMDARs and other postsynaptic scaffolding proteins. ROCK-mediated phosphorylation of SHANK3 in the NAc was essential for spine growth and aversive learning. These findings demonstrate that NMDAR activation initiates a phosphorylation cascade crucial for learning and memory.

Modulation of cortical representations of sensory and contextual information underlies aversive associative learning

Cortical neurons encode both sensory and contextual information, yet it remains unclear how experiences modulate these cortical representations. Here, we demonstrate that trace eyeblink conditioning (TEC), an aversive associative-learning paradigm linking conditioned (CS) with unconditioned stimuli (US), finely tunes cortical coding at both population and single-neuron levels. Initially, we show that the primary somatosensory cortex (S1) is necessary for TEC acquisition, as evidenced by local muscimol administration. At the population level, TEC enhances activity in a small subset (∼20%) of CS- or US-responsive primary neurons (rPNs) while diminishing activity in non-rPNs, including locomotion-tuned or unresponsive PNs. Crucially, TEC learning modulates the encoding of sensory versus contextual information in single rPNs: CS-responsive neurons become less responsive, while US-responsive neurons gain responses to CS. Moreover, we find that the cholinergic pathway, via nicotinic receptors, underlies TEC-induced modulations. These findings suggest that experiences dynamically tune cortical representations through cholinergic pathways.

Direct serotonin release in humans shapes aversive learning and inhibition

The role of serotonin in human behaviour is informed by approaches which allow in vivo modification of synaptic serotonin. However, characterising the effects of increased serotonin signalling in human models of behaviour is challenging given the limitations of available experimental probes, notably selective serotonin reuptake inhibitors. Here we use a now-accessible approach to directly increase synaptic serotonin in humans (a selective serotonin releasing agent) and examine its influence on domains of behaviour historically considered core functions of serotonin. Computational techniques, including reinforcement learning and drift diffusion modelling, explain participant behaviour at baseline and after week-long intervention. Reinforcement learning models reveal that increasing synaptic serotonin reduces sensitivity for outcomes in aversive contexts. Furthermore, increasing synaptic serotonin enhances behavioural inhibition, and shifts bias towards impulse control during exposure to aversive emotional probes. These effects are seen in the context of overall improvements in memory for neutral verbal information. Our findings highlight the direct effects of increasing synaptic serotonin on human behaviour, underlining its role in guiding decision-making within aversive and more neutral contexts, and offering implications for longstanding theories of central serotonin function.

Prior Running, as Well as Swimming, Hinders Swimming‐Based Taste Aversion Learning in Rats

AbstractRunning in an exercise wheel or swimming in a water pool endows rats with a conditioned aversion to a taste substance consumed before the activity. The present study explores the psychophysiological processes underlying these activity‐based taste aversions. Experiment 1 demonstrated that prior experience of running or swimming (i.e., familiarization) hinders subsequent taste aversion learning based on swimming in rats, implying a similar psychophysiological process between running and swimming in establishing taste aversion. Experiment 2 replicated the findings of Experiment 1 with rats obtained from another animal supplier. The running‐to‐swimming cross‐familiarization effect demonstrated in these experiments contrasts with a previous report in which prior swimming had no effect on subsequent running‐based taste aversion learning. Theoretical considerations regarding such asymmetrical cross‐familiarization effects extended to discussions on the similarities and differences in the psychophysiological states induced by running, swimming, and emetic lithium chloride injection.

Nicotine reduces discrimination between threat and safety in the hippocampus, nucleus accumbens and amygdala

Nicotine intake is linked to the maintenance and development of anxiety disorders and impairs adaptive discrimination of threat and safety in rodents and humans. Yet, it is unclear if nicotine exerts a causal pharmacological effect on the affective and neural mechanisms that underlie aversive learning. We conducted a pre-registered, pseudo-randomly and double-blinded pharmacological fMRI study to investigate the effect of acute nicotine on Fear Acquisition and Extinction in non-smokers (n = 88). Our results show that nicotine administration led to decreased discrimination between threat and safety in subjective fear. Nicotine furthermore decreased differential (threat vs. safety) activation in the hippocampus, which was functionally coupled with Nucleus Accumbens and amygdala, compared to placebo controls. Additionally, nicotine led to enhanced physiological arousal to learned threats and overactivation of the ventral tegmental area. This study provides mechanistic evidence that single doses of nicotine impair neural substrates of adaptive aversive learning in line with the risk for the development of pathological anxiety.

An externally validated resting-state brain connectivity signature of pain-related learning

Pain can be conceptualized as a precision signal for reinforcement learning in the brain and alterations in these processes are a hallmark of chronic pain conditions. Investigating individual differences in pain-related learning therefore holds important clinical and translational relevance. Here, we developed and externally validated a novel resting-state brain connectivity-based predictive model of pain-related learning. The pre-registered external validation indicates that the proposed model explains 8-12% of the inter-individual variance in pain-related learning. Model predictions are driven by connections of the amygdala, posterior insula, sensorimotor, frontoparietal, and cerebellar regions, outlining a network commonly described in aversive learning and pain. We propose the resulting model as a robust and highly accessible biomarker candidate for clinical and translational pain research, with promising implications for personalized treatment approaches and with a high potential to advance our understanding of the neural mechanisms of pain-related learning.

Saccharin aversion learning in male and female laboratory rats

The three experiments reported in the present research demonstrated that male rats acquire stronger saccharin aversion than females. Experiment 1 revealed that male rats were better than females in a single-trial saccharin aversion learning with a lithium chloride injection as the unconditioned stimulus (US), when normalizing saccharin intake data. Experiment 2 replicated this finding. The stronger saccharin aversion learning in male rats was also observed in Experiment 3 with wheel running as the US, assessed by the saccharin-vs.-water choice test conducted after six saccharin-running pairing days. Experiments 2 and 3 also showed that the magnitude of induced nausea, reflected in the kaolin clay ingestion, is equivalent between the sexes. Experiment 4 excluded the possibility that the observed sex difference in saccharin aversion is due to the sex difference in inherent saccharin preference. These results, taken together, suggest that potential factors of the sex difference in rats' conditioned taste aversion could include sex disparities in taste perception, the ability to associate taste with nausea, and/or the expression of association learning.

Single dopaminergic neuron DAN-c1 in Drosophila larval brain mediates aversive olfactory learning through D2-like receptors.

The intricate relationship between the dopaminergic system and olfactory associative learning in Drosophila has been an intense scientific inquiry. Leveraging the formidable genetic tools, we conducted a screening of 57 dopaminergic drivers, leading to the discovery of DAN-c1 driver, uniquely targeting the single dopaminergic neuron (DAN) in each brain hemisphere. While the involvement of excitatory D1-like receptors is well-established, the role of D2-like receptors (D2Rs) remains underexplored. Our investigation reveals the expression of D2Rs in both DANs and the mushroom body (MB) of third instar larval brains. Silencing D2Rs in DAN-c1 via microRNA disrupts aversive learning, further supported by optogenetic activation of DAN-c1 during training, affirming the inhibitory role of D2R autoreceptor. Intriguingly, D2R knockdown in the MB impairs both appetitive and aversive learning. These findings elucidate the distinct contributions of D2Rs in diverse brain structures, providing novel insights into the molecular mechanisms governing associative learning in Drosophila larvae.

Neurophysiological and Autonomic Dynamics of Threat Processing During Sustained Social Fear Generalization.

Survival in dynamic environments requires that organisms learn to predict danger from situational cues. One key facet of threat prediction is generalization from a predictive cue to similar cues, ensuring that a cue-outcome contingency is applied beyond the original learning environment. Generalization has been observed in laboratory studies of aversive conditioning: behavioral and physiological processes generalize responses from a stimulus paired with threat (the CS+) to unpaired stimuli, with response magnitudes varying with CS+ similarity. In contrast, work focusing on sensory responses in visual cortex has found a sharpening pattern, in which responses to stimuli closely resembling the CS+ are maximally suppressed, potentially reflecting lateral inhibitory interactions with the CS+ representation. Originally demonstrated with simple visual cues, changes in visuocortical tuning have also been observed in threat generalization learning across facial identities. It is unclear to what extent these visuocortical changes represent transient or sustained effects and if generalization learning requires prior conditioning to the CS+. The present study addressed these questions using EEG and pupillometry in an aversive generalization paradigm involving hundreds of trials using a gradient of facial identities. Visuocortical ssVEP sharpening occurred after dozens of trials of generalization learning without prior differential conditioning, but diminished as learning continued. By contrast, generalization of alpha power suppression, pupil dilation, and self-reported valence and arousal was seen throughout the experiment. Findings are consistent with threat processing models emphasizing the role of changing visucocortical and attentional dynamics when forming, curating, and shaping fear memories as observers continue learning about stimulus-outcome contingencies.

More personal, but not better: The personalization effect in learning neutral and aversive health information

AbstractBackgroundAccording to the personalization effect in multimedia learning, the use of personal and possessive pronouns in instructional materials (e.g., ‘you’ and ‘your’) is beneficial. However, current research suggests that the personalization effect is inverted for emotionally aversive content (e.g., illnesses).ObjectiveThis study investigates whether a beneficial personalization effect can be observed for emotionally neutral health‐related content whereas the effect may be reversed for emotionally aversive health‐related content.MethodsIn this study, 139 university students learned both emotionally aversive learning content on type 1 diabetes (within‐factor) that was presented in either personalized or non‐personalized language (between‐factor). The presentation order of the content (neutral first vs. aversive first) was controlled (between‐factor), resulting in a 2 × 2 × 2 mixed design. The dependent variables measured include learning outcomes (regarding retention and transfer), state anxiety, extraneous cognitive load, motivation and learning time.Results and ConclusionsIn the transfer test, learners generally performed better when learning with non‐personalized instructional materials than with personalized instructional materials, regardless of whether the content was emotionally neutral or aversive. The results raise questions regarding the robustness of the personalization effect and the underlying mechanisms of the inverted personalization effect. An alternative explanation to be investigated is whether the direct reference to a disease that the participants do not have (here: ‘your type 1 diabetes’) leads to schema interference, which could be responsible for poorer learning performance—even if the learning content about the disease can be considered emotionally neutral.

Compensatory enhancement of input maintains aversive dopaminergic reinforcement in hungry Drosophila

Hungry animals need compensatory mechanisms to maintain flexible brain function, while modulation reconfigures circuits to prioritize resource seeking. In Drosophila, hunger inhibits aversively reinforcing dopaminergic neurons (DANs) to permit the expression of food-seeking memories. Multitasking the reinforcement system for motivation potentially undermines aversive learning. We find that chronic hunger mildly enhances aversive learning and that satiated-baseline and hunger-enhanced learning require endocrine adipokinetic hormone (AKH) signaling. Circulating AKH influences aversive learning via its receptor in four neurons in the ventral brain, two of which are octopaminergic. Connectomics revealed AKH receptor-expressing neurons to be upstream of several classes of ascending neurons, many of which are presynaptic to aversively reinforcing DANs. Octopaminergic modulation of and output from at least one of these ascending pathways is required for shock- and bitter-taste-reinforced aversive learning. We propose that coordinated enhancement of input compensates for hunger-directed inhibition of aversive DANs to preserve reinforcement when required.