Human Fear Conditioning Research Articles

Complete the triangulation: Quantifying differential fear conditioning with a noninterfering and sensitive behavioral measure along with self‐report and physiological measures

According to the multicomponent view, emotion is expressed through subjective feelings and thoughts, physiological activation, and behavioral responses. In human fear conditioning research, the former two are much more popular than the third category. One concern is that concurrent behavioral probes may interfere with the conditioning process. To allow triangulation of emotion research through simultaneous employment of subjective, physiological, and behavioral measurement, it is necessary to find behavioral measures that meet the criteria of causing no interference while being sensitive to conditioning. In this study, a basic visual attention task was examined in terms of its impact on differential fear conditioning as measured by both subjective (i.e., self-reported fear and shock estimation) and physiological (i.e., skin conductance response/SCR) expression; and its ability to detect fear conditioning indicated by a reaction time (RT) or accuracy difference between the two conditioned stimuli (CS+ vs. CS-). While participants in the probe group (n=86) completed differential fear conditioning with the behavioral task, those in the no-probe group (n=76) underwent conditioning by itself. Based on self-reported fear, shock estimation, and SCR, both groups successfully acquired differential fear with no apparent between-group difference in the degree of conditioning. In the probe group, RT but not accuracy exhibited a difference between CS+ and CS-. These findings suggest that the selected visual attention task does not interfere with differential fear conditioning measured via SCR and self-report and is a sensitive measure of differential conditioning. Exploratory individual analyses also revealed significant relationships between the above measures.

The Role of Contingency Awareness in Human Fear Conditioning: Electrophysiological Analysis

The recognition of the conditioned-unconditioned stimulus (CS-US) association in classical conditioning is referred to as contingency awareness. The neural underpinnings of contingency awareness in human fear conditioning are poorly understood.

Neural underpinnings of contingency awareness in human fear conditioning

IntroductionThe recognition of the conditioned-unconditioned stimulus (CS-US) association in classical conditioning is referred to as contingency awareness. The neural underpinnings of contingency awareness in human fear conditioning are poorly understood.ObjectivesWe aimed to explore the EEG correlates of contingency awareness.MethodsHere, we recorded electroencephalography (EEG) from a sample of 20 participants in a semantic conditioning experiment. In the acquisition phase the participants were presented with sequences of words from two semantic categories paired with tactile stimulation followed by presentation of a neutral sound (US-) ((e.g., animals -> left hand vibration -> US-, clothes -> right hand vibration -> US-). In the test phase the association violated in 50% of trials which followed by a presentation of a loud noise (US+). The participants were only instructed to listen carefully. On the basis of self-reported contingency awareness, twenty participants were divided in aware (N=12) and unaware (N=8) group.ResultsThe aware group expressed a non-lateralized effect of alpha-beta (12-23 Hz) suppression along with a more negative CNV at central channels preceding presentation of the vibration (main effect of Group). Also, CNV was more negative in expectation of US+ comparing with expectation of US- in the aware group but not in the unaware group.ConclusionsThe results indicate that contingency awareness is accompanied by neural patterns reflecting expectation as can be seen in the suppression of somatosensory alpha-beta activity before expected presentation of the vibration as well as in CNV in expectation of an aversive event.

The effects of sleep and wakefulness on human fear conditioning

IntroductionStudies on fear conditioning have made important contributions to the understanding of affective learning mechanisms as well as its applications (e.g., anxiety disorders, post-traumatic stress disorder). However, central mechanisms of sleep related consolidation of fear memory in humans have been almost neglected by previous studies.ObjectivesIn the current study we aimed to test effects of sleep and a period wakefulness on fear conditioned responses.MethodsIn our experiment in a group 18 healthy volunteers event-related brain potentials (ERP), heart rate variability (HRV) and behavioral responses were recorded during a fear conditioning procedure presented twice, before daytime sleep (2h) or control intervention (a period of wakefulness) and after. The conditioning procedure involved pairing of a neutral tone (CS+) with a highly unpleasant sound (UCS+).ResultsDifferential conditioning manifested itself in the contingent negative variance (CNV)-like slow ERP component. Both period of sleep and wakefulness resulted in an increased amplitude of the CNV to CS+. But we did not find an interaction effect of Time (Pre-Post) by Intervention (Sleep-Wake), suggesting that sleep did not affect the conditioned response differently as compared to a period of wakefulness. An apparent increase in HRV after a period of wakefulness did not affect fear conditioned responses (CNV and valence ratings).ConclusionsTo summarize, the data indicate that fear memories are consolidated with the course of time with no beneficial effect of sleep; relearning of fear causes stronger differential responses as measured by slow wave amplitude but not behavior; increase of HRV does not affect fear learning.

Attentive immobility in the face of inevitable distal threat-Startle potentiation and fear bradycardia as an index of emotion and attention.

During fear conditioning, a cue (CS) signals an inevitable distal threat (US) and evokes a conditioned response that can be described as attentive immobility (freezing). The organism remains motionless and monitors the source of danger while startle responses are potentiated, indicating a state of defensive hypervigilance. Although in animals vagally mediated fear bradycardia is also reliably observed under such circumstances, results are mixed in human fear conditioning. Using a single-cue fear conditioning and extinction protocol, we tested cardiac reactivity and startle potentiation indexing low-level defensive strategies in a fear-conditioned (n=40; paired presentations of CS and US) compared with a non-conditioned control group (n=40; unpaired presentations of CS and US). Additionally, we assessed shock expectancy ratings on a trial-by-trial basis indexing declarative knowledge of the previous contingencies. Half of each group underwent extinction under sham or active transcutaneous vagus nerve stimulation (tVNS), serving as additional proof of concept. We found stronger cardiac deceleration during CS presentation in the fear learning relative to the control group. This learned fear bradycardia was positively correlated with conditioned startle potentiation but not with declarative knowledge of CS-US contingencies. TVNS abolished differences in heart rate changes between both groups and removed the significant correlation between late cardiac deceleration and startle potentiation in the fear learning group. Results suggest, fear-conditioned cues evoke attentive immobility in humans, characterized by cardiac deceleration and startle potentiation. Such defensive response pattern is elicited by cues predicting inevitable distal threat and resembles conditioned fear responses observed in rodents.

A dimensional measure of safety behavior: A non-dichotomous assessment of costly avoidance in human fear conditioning

Safety behavior prevents the occurrence of threat, thus it is typically considered adaptive. However, safety behavior in anxiety-related disorders is often costly, and persists even the situation does not entail realistic threat. Individuals can engage in safety behavior to varying extents, however, these behaviors are typically measured dichotomously (i.e., to execute or not). To better understand the nuances of safety behavior, this study developed a dimensional measure of safety behavior that had a negative linear relationship with the admission of an aversive outcome. In two experiments, a Reward group receiving fixed or individually calibrated incentives competing with safety behavior showed reduced safety behavior than a Control group receiving no incentives. This allowed extinction learning to a previously learnt warning signal in the Reward group (i.e., updating the belief that this stimulus no longer signals threat). Despite the Reward group exhibited extinction learning, both groups showed a similar increase in fear to the warning signal once safety behavior was no longer available. This null group difference was due to some participants in the Reward group not incentivized enough to disengage from safety behavior. Dimensional assessment revealed a dissociation between low fear but substantial safety behavior to a safety signal in the Control group. This suggests that low-cost safety behavior does not accurately reflect the fear-driven processes, but also other non-fear-driven processes, such as cost (i.e., engage in safety behavior merely because it bears little to no cost). Pinpointing both processes is important for furthering the understanding of safety behavior.

Human fear conditioning is moderated by stimulus contingency instructions

Recent research findings indicate that human fear conditioning is affected by instructions, particularly those concerning the contingency between the conditioned stimulus (CS) and the unconditioned stimulus (US). However, whether or not such instructions were provided to participants often remains unsaid in fear conditioning studies. In the current study (N = 102), we investigated whether conditioned fear acquisition depends on CS-US contingency instructions. Participants were randomly assigned to one of three groups. The first group was instructed about the precise CS-US contingency before conditioning. The second group was instructed to discover the CS-US contingency. The third group did not receive any contingency instructions. We found facilitated fear acquisition (using skin conductance and startle) and increased contingency awareness in the first and second group compared to the third group. Furthermore, contingency reversal instructions immediately reversed conditioned responses. Based on these results, we advise to systematically report the contingency instructions used in fear conditioning research.

Learning dynamics of electrophysiological brain signals during human fear conditioning

Electrophysiological studies in rodents allow recording neural activity during threats with high temporal and spatial precision. Although fMRI has helped translate insights about the anatomy of underlying brain circuits to humans, the temporal dynamics of neural fear processes remain opaque and require EEG. To date, studies on electrophysiological brain signals in humans have helped to elucidate underlying perceptual and attentional processes, but have widely ignored how fear memory traces evolve over time. The low signal-to-noise ratio of EEG demands aggregations across high numbers of trials, which will wash out transient neurobiological processes that are induced by learning and prone to habituation. Here, our goal was to unravel the plasticity and temporal emergence of EEG responses during fear conditioning. To this end, we developed a new sequential-set fear conditioning paradigm that comprises three successive acquisition and extinction phases, each with a novel CS+/CS- set. Each set consists of two different neutral faces on different background colors which serve as CS+ and CS-, respectively. Thereby, this design provides sufficient trials for EEG analyses while tripling the relative amount of trials that tap into more transient neurobiological processes. Consistent with prior studies on ERP components, data-driven topographic EEG analyses revealed that ERP amplitudes were potentiated during time periods from 33–60 ms, 108–200 ms, and 468–820 ms indicating that fear conditioning prioritizes early sensory processing in the brain, but also facilitates neural responding during later attentional and evaluative stages. Importantly, averaging across the three CS+/CS- sets allowed us to probe the temporal evolution of neural processes: Responses during each of the three time windows gradually increased from early to late fear conditioning, while long-latency (460–730 ms) electrocortical responses diminished throughout fear extinction. Our novel paradigm demonstrates how short-, mid-, and long-latency EEG responses change during fear conditioning and extinction, findings that enlighten the learning curve of neurophysiological responses to threat in humans.

Fear in the context of pain: Lessons learned from 100 years of fear conditioning research

Human fear conditioning research since Watson's case study on “Little Albert” has vastly evolved and its impact today is reaching far beyond phobic anxiety. This review focuses on how fear conditioning research, mainly using exteroceptive conditioned stimuli (CSs) and aversive, non-noxious stimuli as unconditioned stimuli (USs), has been extended and translated to chronic pain research. We describe the different pain-related fear conditioning paradigms using proprioceptive and interoceptive CSs and painful stimuli as USs that have been developed to study specific forms of pain-related fear (i.e. fear of movement, fear of touch, fear of visceral sensations, and fear of penetration) that are relevant for different chronic pain conditions (i.e. musculoskeletal pain, neuropathic pain, visceral pain, and genital pain). We present evidence that patients with chronic pain demonstrate impaired safety learning and excessive fear generalization; learning anomalies that have also been observed in anxiety disorders. Extinction-based protocols (exposure in vivo) have been developed to reduce pain-related fear and increase daily functioning in various chronic pain disorders. Finally, we outline some challenges and future directions to further our understanding of learning mechanisms underlying the development, persistence, and treatment of chronic pain disability.

Navigating the garden of forking paths for data exclusions in fear conditioning research.

In this report, we illustrate the considerable impact of researcher degrees of freedom with respect to exclusion of participants in paradigms with a learning element. We illustrate this empirically through case examples from human fear conditioning research, in which the exclusion of 'non-learners' and 'non-responders' is common - despite a lack of consensus on how to define these groups. We illustrate the substantial heterogeneity in exclusion criteria identified in a systematic literature search and highlight the potential problems and pitfalls of different definitions through case examples based on re-analyses of existing data sets. On the basis of these studies, we propose a consensus on evidence-based rather than idiosyncratic criteria, including clear guidelines on reporting details. Taken together, we illustrate how flexibility in data collection and analysis can be avoided, which will benefit the robustness and replicability of research findings and can be expected to be applicable to other fields of research that involve a learning element.

Human fear conditioning: From neuroscience to the clinic

Both clinicians and neuroscientists have been long interested in the topic of fear conditioning, with recent advances in neuroscience, in particular, igniting a shared interest in further translation between these domains. Here, we review some historical aspects of this relationship and the progress that has been made in translating the neuroscientific study of fear conditioning to the conceptualization and treatment of mental disorders, especially anxiety-related disorders. We also address some conceptual and methodological challenges faced by this research, and offer some suggestions to support future progress in the field.

Sleep deprivation increases threat beliefs in human fear conditioning.

SummarySleep disturbances and anxiety disorders exhibit high comorbidity levels, but it remains unclear whether sleep problems are causes or consequences of increased anxiety. To experimentally probe the aetiological role of sleep disturbances in anxiety, we investigated in healthy participants how total sleep deprivation influences fear expression in a conditioning paradigm. In a fear conditioning procedure, one face stimulus (conditioned stimulus [CS+]) was paired with electric shock, whereas another face stimulus was not (unpaired stimulus [CS−]). Fear expression was tested the next morning using the two face stimuli from the training phase and a generalization stimulus (i.e. a morph between the CS+ and CS− stimuli). Between fear conditioning and test, participants were either kept awake in the laboratory for 12 hr (n = 20) or had a night of sleep at home (n = 20). Irrespective of stimulus type, subjective threat expectancies, but not skin conductance responses, were enhanced after sleep deprivation, relative to regular sleep. These results suggest that sleep disturbances may play a role in anxiety disorders by increasing perceived threat.

Insensitivity of auditory mismatch negativity to classical fear conditioning and extinction in healthy humans.

The relationship between auditory mismatch negativity (MMN) and the neural cognitive processes of fear has been suggested in both healthy participants and patients with fear-related mental disorders such as post-traumatic stress disorder and panic disorder. The present study sought to confirm whether the MMN is affected by classical fear conditioning in healthy participants. MMN amplitude, N1 amplitude, and skin conductance level (SCL) in 20 healthy volunteers during a fear-conditioning paradigm consisting of three phases (habituation, fear acquisition, and fear extinction) were recorded. Red and blue light signals were presented as the conditioned stimuli CS+ (threat cue) and CS- (safety cue), respectively. In addition, an aversive electrical stimulus was delivered as the unconditioned stimulus with CS+ in the fear-acquisition phase. No MMN amplitude changes were observed between the CS types during the three phases. In the acquisition phase, the mean SCL during CS+ was significantly higher than that during CS-. The MMN amplitude and deviant N1 amplitude in the extinction phase were significantly lower than those in the other phases regardless of the CS type. Despite the clear alteration of SCL between CS types in the acquisition phase, no significant differences in MMN were observed. Decreased MMN and deviant N1 in the fear-extinction phase were considered to be mainly due to decreased arousal or attention level. Results indicate that the auditory MMN amplitude was not affected by the cognitive process of fear recognized by other sense modalities.

When translational neuroscience fails in the clinic: Dexamethasone prior to virtual reality exposure therapy increases drop-out rates

Posttraumatic stress disorder (PTSD) is characterized by exaggerated expression of fear responses to danger and safety cues. Translational research suggests that dexamethasone facilitates fear extinction in animal and human fear conditioning models. For this randomized, placebo-controlled trial (N = 27), we aimed to translate these findings to the clinic by using virtual reality exposure (VRE) therapy for OEF/OIF/OND veterans with PTSD to determine whether dexamethasone will increase the efficacy of exposure therapy for VRE relative to placebo. VRE sessions involved imaginal exposure to the most traumatic war memories while viewing a computer-generated view of virtual Iraq or Afghanistan with multisensory stimulus options used to match patient’s description of the trauma. VRE was effective in reducing PTSD symptoms but there was no interaction with dexamethasone. Drop-out rate was significantly higher in the dexamethasone group, with 10 of 13 (76.9%) participants in this group discontinuing, compared to only 4 of 14 (28.5%) in the placebo group, χ2 = 6.31, p = 0.02. Results indicate that the dexamethasone group may have experienced an increase in PTSD symptoms, particularly re-experiencing, at session 2 following first drug administration. Contrary to study hypotheses, dexamethasone did not enhance exposure therapy outcomes and was associated with increased drop-out. This demonstrates potential pitfalls in translating neuroscience models to the clinic; future research carefully examining glucocorticoid mechanisms involved in therapy augmentation is warranted.

Critical evaluation of current data analysis strategies for psychophysiological measures of fear conditioning and extinction in humans.

Fear conditioning and extinction is a construct integral to understanding trauma-, stress- and anxiety-related disorders. In the laboratory, associative learning paradigms that pair aversive with neutral stimuli are used as analogues to real-life fear learning. These studies use physiological indices, such as skin conductance, to sensitively measure rates and intensity of learning and extinction. In this review, we discuss some of the potential limitations in interpreting and analysing physiological data during the acquisition or extinction of conditioned fear. We argue that the utmost attention should be paid to the development of modelling approaches of physiological data in associative learning paradigms, by illustrating the lack of replicability and interpretability of results in current methods. We also show that statistical significance may be easily achieved in this paradigm without more stringent data and data analysis reporting requirements, leaving this particular field vulnerable to misleading conclusions. This review is written so that issues and potential solutions are accessible to researchers without mathematical training. We conclude the review with some suggestions that all laboratories should be able to implement, including visualising the full data set in publications and adopting modelling, or at least regression-based, approaches.

Secondary extinction reduces reinstatement of threat expectancy and conditioned skin conductance responses in human fear conditioning

Background and objectivesSecondary extinction refers to the phenomenon that extinction of one conditioned stimulus (CS) results in the reduction of conditioned responses for other CSs conditioned with the same unconditioned stimulus (US). Previous research with rats has demonstrated that secondary extinction can interfere with the return of conditioned fear after a reinstatement manipulation. Here we investigated this phenomenon in two pre-registered studies in humans. MethodIn both experiments, distinct CSs were paired with an electrical stimulation. Next, conditioned reactions to both CSs were extinguished and thereafter reinstated through the administration of three unsignaled electrical stimulations. Crucially, before participants continued with the reinstatement test, half of the participants received secondary extinction trials whereas the other half did not receive these trials. ResultsOur results indicate that secondary extinction reduced reinstatement of threat expectancies and skin conductance responses, but the effect on skin conductance was only found in the second experiment. LimitationsThe studies were conducted in a laboratory setting with healthy students. Additional research will be required to determine the feasibility of applying secondary extinction in a (sub)clinical context. ConclusionsTo our knowledge, this is the first demonstration of secondary extinction and its effect on reinstatement of conditioned fear in humans. We relate our findings to the earlier research with rats and discuss their relevance for exposure therapy.

The influence of acoustic startle probes on fear learning in humans

Even though human fear-conditioning involves affective learning as well as expectancy learning, most studies assess only one of the two distinct processes. Commonly used read-outs of associative fear learning are the fear-potentiated startle reflex (FPS), pupil dilation and US-expectancy ratings. FPS is thought to reflect the affective aspect of fear learning, while pupil dilation reflects a general arousal response. However, in order to measure FPS, aversively loud acoustic probes are presented during conditioning, which might in itself exert an effect on fear learning. Here we tested the effect of startle probes on fear learning by comparing brain activation (fMRI), pupil dilation and US-expectancy ratings with and without acoustic startle probes within subjects. Regardless of startle probes, fear conditioning resulted in enhanced dACC, insula and ventral striatum activation. Interaction analyses showed that startle probes diminished differential pupil dilation between CS+ and CS− due to increased pupil responses to CS−. A trend significant interaction effect was observed for US-expectancy and amygdala activation. Startle probes affect differential fear learning by impeding safety learning, as measured with pupil dilation, a read-out of the cognitive component of fear learning. However, we observed no significant effect of acoustic startle probes on other measures of fear learning.

Measuring the conditioned response: A comparison of pupillometry, skin conductance, and startle electromyography

In human fear conditioning studies, different physiological readouts can be used to track conditioned responding during fear learning. Commonly employed readouts such as skin conductance responses (SCR) or startle responses have in recent years been complemented by pupillary readouts, but to date it is unknown how pupillary readouts relate to other measures of the conditioned response. To examine differences and communalities among pupil responses, SCR, and startle responses, we simultaneously recorded pupil diameter, skin conductance, and startle electromyography in 47 healthy subjects during fear acquisition, extinction, and a recall test on 2 consecutive days. The different measures correlated only weakly, displaying most prominent differences in their response patterns during fear acquisition. Whereas SCR and startle responses habituated, pupillary measures did not. Instead, they increased in response to fear conditioned stimuli and most closely followed ratings of unconditioned stimulus (US) expectancy. Moreover, we observed that startle-induced pupil responses showed stimulus discrimination during fear acquisition, suggesting a fear potentiation of the auditory pupil reflex. We conclude that different physiological outcome measures of the conditioned response inform about different cognitive-affective processes during fear learning, with pupil responses being least affected by physiological habituation and most closely following US expectancy.

Temporal context cues in human fear conditioning: Unreinforced conditional stimuli can segment learning into distinct temporal contexts and drive fear responding

In associative learning, if stimulus A is presented in the same temporal context as the conditional stimulus (CS) - outcome association (but not in a way that allows an A–CS association to form) it becomes a temporal context cue, acquiring the ability to activate this context and retrieve the CS-outcome association. We examined whether a CS- presented during acquisition or extinction that predicted the absence of the unconditional stimulus (US) could act as a temporal context cue, reducing or enhancing responding, in differential fear conditioning. Two groups received acquisition (CSx–US, CSa–noUS) in phase 1 and extinction (CSx–noUS; CSe–noUS) in phase 2 (AE groups), and two groups received extinction in phase 1 and acquisition in phase 2 (EA groups). After a delay, participants were presented with either CSa (AEa and EAa groups) or CSe (AEe and EAe groups). Responding to CSx was enhanced after presentation of CSa but reduced after presentation of CSe, suggesting that training was segmented into two learning episodes and that the unreinforced CS present during an episode retrieved the CSx–US or CSx–noUS association. These findings suggest that temporal context cues may enhance or reduce fear responding, providing an exciting new avenue for relapse prevention research.

Effects of bright light exposure on human fear conditioning, extinction, and associated prefrontal activation.

Bright light (BL) not only regulates human emotion and circadian physiology but can also directly modulate emotional memories. Impaired fear extinction and enhanced fear acquisition and consolidation are hallmarks of fear-circuitry disorders; thus, we tested whether BL facilitates fear extinction and inhibits fear acquisition. We randomly exposed 29 healthy humans to high- (9000 lx) or low-intensity light (<500 lx) for 15 min, near the nadir of the phase response to light, in a single-blind manner. Simultaneously with the light exposure, subjects performed fear extinction training and second fear acquisition, where a visual conditioned stimulus (CS), previously paired with an electric shock unconditioned stimulus (US), was presented without the US, while another CS was newly paired with the US. Conditioned responses (CRs) and changes in prefrontal cortex (PFC) activity were determined during encoding and delayed recall sessions. BL-exposed subjects exhibited lower extinction-related PFC activity and marginally higher acquisition-related PFC activity during light exposure than subjects exposed to control light. Twenty-four hours later, BL reduced CRs to both the extinguished and non-extinguished CSs with marginally lower extinction-related PFC activation, suggesting that BL enhanced fear extinction, while suppressing fear acquisition. Further, BL sustained tolerance to fear re-conditioning. Our results demonstrate that a single and brief BL exposure, synchronized with fear extinction and acquisition, instantaneously influences prefrontal hemodynamic responses and alleviates fear expression after 24 h. Although the specificity of BL effects deems further investigation, our findings indicate the clinical relevance of adjunctive BL intervention in exposure-based cognitive-behavioral therapy for fear-circuitry disorders.