Threat Memory Research Articles

A Ventromedial Prefrontal-to-Lateral Entorhinal Cortex Pathway Modulates the Gain of Behavioral Responding During Threat

BackgroundThe ability to correctly associate cues and contexts with threat is critical for survival, and the inability to do so can result in threat-related disorders such as posttraumatic stress disorder. The prefrontal cortex (PFC) and hippocampus are well known to play critical roles in cued and contextual threat memory processing. However, the circuits that mediate prefrontal-hippocampal modulation of context discrimination during cued threat processing are less understood. Here, we demonstrate the role of a previously unexplored projection from the ventromedial region of PFC (vmPFC) to the lateral entorhinal cortex (LEC) in modulating the gain of behavior in response to contextual information during threat retrieval and encoding. MethodsWe used optogenetics followed by in vivo calcium imaging in male C57/B6J mice to manipulate and monitor vmPFC-LEC activity in response to threat-associated cues in different contexts. We then investigated the inputs to, and outputs from, vmPFC-LEC cells using Rabies tracing and channelrhodopsin-assisted electrophysiology. ResultsvmPFC-LEC cells flexibly and bidirectionally shaped behavior during threat expression, shaping sensitivity to contextual information to increase or decrease the gain of behavioral output in response to a threatening or neutral context, respectively. ConclusionsGlutamatergic vmPFC-LEC cells are key players in behavioral gain control in response to contextual information during threat processing and may provide a future target for intervention in threat-based disorders.

'Cut the roots': rTMS reveals the crucial role of prefrontal cortex in threat memory consolidation in humans

'Cut the roots': rTMS reveals the crucial role of prefrontal cortex in threat memory consolidation in humans

Threat learning impairs subsequent associative inference

Despite it being widely acknowledged that the most important function of memory is to facilitate the prediction of significant events in a complex world, no studies to date have investigated how our ability to infer associations across distinct but overlapping experiences is affected by the inclusion of threat memories. To address this question, participants (n = 35) encoded neutral predictive associations (A → B). The following day these memories were reactivated by pairing B with a new aversive or neutral outcome (B → CTHREAT/NEUTRAL) while pupil dilation was measured as an index of emotional arousal. Then, again 1 day later, the accuracy of indirect associations (A → C?) was tested. Associative inferences involving a threat learning memory were impaired whereas the initial memories were retroactively strengthened, but these effects were not moderated by pupil dilation at encoding. These results imply that a healthy memory system may compartmentalize episodic information of threat, and so hinders its recall when cued only indirectly. Malfunctioning of this process may cause maladaptive linkage of negative events to distant and benign memories, and thereby contribute to the development of clinical intrusions and anxiety.

Opto-extinction of a threat memory in mice

Memories of past experiences guide future behaviour. Sparse ensembles of neurons, known as engrams, are thought to store memories in the brain. Neurons involved in a particular engram (“engram neurons”) are necessary for subsequent memory expression as memory retrieval is thought to be initiated by an external sensory cue reactivating engram neurons. However, conditions or environments are dynamic, such that future behaviour should be flexible. The role of engrams in mediating flexible behaviour is not understood. Here we examined this question using one type of flexible behaviour, extinction of a threat response. An initially neutral tone is first paired with an aversive footshock such that the tone alone induces defensive freezing. After subsequent repeated tone presentations without the footshock, rodents no longer freeze to the tone. Because the tone cue is thought to reactivate the engram to induce memory retrieval, we examined whether it is possible to induce an extinction-like behavioural effect by optogenetically reactivating the lateral amygdala component of the engram alone (without tone re-exposure). Similar to tone-induced extinction, mice showed decreased freezing to optogenetic stimulation of the lateral amygdala engram in the “extinction training” session. Moreover, “opto-extinguished” mice showed decreased freezing to the tone when subsequently tested for retrieval of the extinction training in the same context, suggesting that the opto-extinction transferred to the actual sensory stimulus. However, unlike tone extinction, in which mice showed renewal of tone-induced freezing when tested in a novel context, opto-extinguished mice continued to show a deficit in tone-induced freezing. Extinction has been characterized as new learning that inhibits the original memory or a phenomenon in which the original memory is "unlearned". Our findings suggest that opto-extinction may silence the original engram to “unlearn” the original memory.

Facilitated extinction of conditioned fear responses by delta 9-tetrahyrdrocannabidol in humans: a pilot study.

We sought to determine whether acute delta 9-tetrahyrdrocannabidol (THC) administration would facilitate fear extinction in young occasional cannabis users, given that animal models indicate THC facilitates extinction learning, and recent studies indicate THC administration may also enhance threat memory extinction in humans. On each of the 2days, 24+hour THC-deprived participants were conditioned to fear visual stimuli in a delay conditioning and extinction paradigm. Both CS+ and CS- were faces of negative emotional valence, with the CS+ paired with mild electric shock. Throughout both conditioning and extinction paradigms, EEG was measured to quantify event-related potentials for these learning processes. Following conditioning, individuals, in a randomized and counter-balanced order, smoked either an active THC cigarette (26.25mg/2.7% THC) or a placebo marijuana cigarette (0.002% THC) on 1day and the opposite cigarette on the second day. After smoking, CS+ and CS- were presented without shock, resulting in extinction of conditioned fear. Relative to placebo, THC facilitated extinction of the conditioned response to the CS+, as reflected by reductions in late positive potential amplitude during extinction learning. The results indicate that acute THC administration may facilitate extinction of the conditioned fear response in humans.

Pavlovian threat conditioning can generate intrusive memories that persist over time

Although Pavlovian threat conditioning has proven to be a useful translational model for the development of anxiety disorders, it remains unknown if this procedure can generate intrusive memories – a symptom of many anxiety-related disorders, and whether intrusions persist over time. Social support has been related to better adjustment after trauma however, experimental evidence regarding its effect on the development of anxiety-related symptoms is sparse. We had two aims: to test whether threat conditioning generates intrusive memories, and whether different social support interactions impacted expression of emotional memories. Non-clinical participants (n = 81) underwent threat conditioning to neutral stimuli. Participants were then assigned to a supportive, unsupportive, or no social interaction group, and asked to report intrusive memories for seven days. As predicted, threat conditioning can generate intrusions, with greater number of intrusions of CS+ (M = 2.35, SD = 3.09) than CS- (M = 1.39, SD = 2.17). Contrary to predictions, compared to no social interaction, supportive social interaction did not reduce, and unsupportive interaction did not increase skin conductance of learned threat or number of intrusions. Unsupportive interaction resulted in a relative difference in number of intrusions to CS + vs CS-, suggesting that unsupportive interaction might have increased image-based threat memories. Intrusions were still measurable one year after conditioning (one-year follow-up; n = 54), when individuals with higher trait anxiety and greater number of previous trauma experiences reported more intrusions. Our findings show that threat conditioning can create long-lasting intrusions, offering a novel experimental psychopathology model of intrusive memories with implications for both research on learning and clinical applications.

Impact of cannabidiol-rich hemp extract oil on reconsolidation disruption of naturalistic interoceptive aversive memory in humans: Protocol for a randomized clinical trial

BackgroundPreclinical experiments with rodents demonstrate that cannabidiol (CBD), the non-psychotomimetic constituent of the Cannabis sativa plant, disrupts reconsolidation of aversive memories conditioned in the laboratory when administered within the memory reconsolidation window (< 6 h. post-retrieval) by indirectly activating cannabinoid type-1 (CB1) receptors in the dorsal anterior cingulate cortex (dACC). Based on these findings, we aim to test whether administration of 300 mg CBD-rich hemp extract oil following fear reactivation of an aversive interoceptive threat memory can disrupt reconsolidation of naturalistic aversive memories in humans. More specifically, naturalistic interoceptive aversive memories, a form of transdiagnostic fear memory that contributes to the pathogenesis of fear-related disorders such as panic disorder, posttraumatic stress disorder (PTSD), and illness anxiety disorder. MethodsFor this proof-of-concept, placebo-controlled double-blind trial, volunteers (n = 99) reporting elevated fears of somatic sensations will be stratified on biological sex and randomized to one of three intervention arms: (a). CBD-rich oil administered within the reconsolidation window, (b) Placebo oil administered within the reconsolidation window; or (c) CBD-rich oil administered outside of the reconsolidation window. Change in emotional reactivity to a 35% CO2 challenge from baseline to two-week follow-up will serve as our primary outcome. ConclusionStudy findings may contribute towards the development of a novel brief transdiagnostic intervention guided by reconsolidation theory for individuals prone to fear-related psychiatric disorders.Trial Registration: ClinicalTrials.gov Identifier: NCT04726475

Highly Demand Working Memory Intervention Weakens a Reactivated Threat Memory and the Associated Cognitive Biases

Anxiety disorders are the most frequent type of mental disorder. Threat-conditioning memory plays a central role in anxiety disorders, impacting complex cognitive systems by modifying behavioral responses to fearful stimuli and inducing an overestimation of potential threats. Here, we analyzed the reminder-dependent amnesia on physiological responses, unconditioned stimulus (US) expectancy ratings, and measures of cognitive bias towards the threat of a threat-conditioning memory.Subjects received differential threat-conditioning. Twenty-four hours later, after reactivation of the memory of threat-conditioning, one group performed a high demand working memory task (HWM) and a second group a low demand working memory task (LWM). A third group only performed the HWM task. Retention of conditioned threat memory was tested on Day 3 in an extinction session followed by a reinstatement test. Tasks targeting stimulus representation, valuation, and attentional bias towards threat were performed. We show that the reminder-dependent intervention with an HWM weakened memory retention as expressed in skin conductance response (SCR) and faded the representation and valuation towards the threat, but it did not affect US expectancy or attentional bias. Our findings provide evidence for the experimental psychopathology approach opening the possibility to weaken both Threat conditioning memory and the systems associated with the maintenance of anxiety features.

Frequency-Dependent Plasticity in the Temporal Association Cortex Originates from the Primary Auditory Cortex, and Is Modified by the Secondary Auditory Cortex and the Medial Geniculate Body.

The brain areas that mediate the formation of auditory threat memory and perceptual decisions remain uncertain to date. Candidates include the primary (A1) and secondary (A2) auditory cortex, the medial division of the medial geniculate body (MGm), amygdala, and the temporal association cortex. We used chemogenetic and optogenetic manipulations with in vivo and in vitro patch-clamp recordings to assess the roles of these brain regions in threat memory learning in female mice. We found that conditioned sound (CS) frequency-dependent plasticity resulted in the formation of auditory threat memory in the temporal association cortex. This neural correlated auditory threat memory depended on CS frequency information from A1 glutamatergic subthreshold monosynaptic inputs, CS lateral inhibition from A2 glutamatergic disynaptic inputs, and non-frequency-specific facilitation from MGm glutamatergic monosynaptic inputs. These results indicate that the A2 and MGm work together in an inhibitory-facilitative role.SIGNIFICANCE STATEMENT: The ability to recognize specific sounds to avoid predators or seek prey is a useful survival tool. Improving this ability through experiential learning is an added advantage requiring neural plasticity. As an example, humans must learn to distinguish the sound of a car horn, and thus avoid oncoming traffic. Our research discovered that the temporal association cortex can encode this kind of auditory information through tonal receptive field plasticity. In addition, the results revealed the underlying synaptic mechanisms of this process. These results extended our understanding of how meaningful auditory information is processed in an animal's brain.

Sleep Deprivation During Memory Consolidation, but Not Before Memory Retrieval, Widens Threat Generalization to New Stimuli.

Past aversive experiences shape our ability to deal with future dangers, through the encoding of implicit and explicit memory traces and through the ability to generalize defensive reactions to new stimuli resembling learned threats. Numerous evidence demonstrate that sleep is important for the consolidation of memories related to threatening events. However, there is a lack of studies examining the effects of sleep deprivation on the retrieval of consolidated threat memories, and previous studies on the role of sleep in threat generalization have produced mixed results. To address these issues, here we adopted a differential threat conditioning and a delayed (second half of the night) sleep deprivation during the first or the seventh night after learning. We found no effects of sleep deprivation on either implicit or explicit threat memories, regardless of its occurrence timing. Conversely, implicit but not explicit responses to novel cues similar to a learned threat displayed a widened generalization pattern, but only if sleep deprivation took place during the first night after conditioning and not if it occurred during the seventh night after conditioning. Therefore, we propose that sleeping after exposure to danger may support optimal implicit discrimination processes to evaluate new signals in the future and that even a brief period of sleeplessness may widen threat generalization to new stimuli, which is a hallmark of several threat-related disorders.

P109. Backward Conditioning Indirectly Retrieves Threat Memories: Implications for Internalizing Psychopathology

In a therapeutic context, extinguishing contextual threat memories can be difficult, as direct exposure to the threatening context is often inaccessible, thus these memories must be indirectly retrieved. Unlike forward-conditioned (FW) cues, where an unconditioned stimulus (US) follows the cue, backward-conditioned (BW) cues are presented immediately after the US. BW cues do not become directly associated with the US, but may indirectly retrieve threat memories via activation of the conditioning context.

No evidence for disruption of reconsolidation of conditioned threat memories with a cognitively demanding intervention

Simultaneous execution of memory retrieval and cognitively demanding interventions alter the subjective experience of aversive memories. This principle can be used in treatment to target traumatic memories. An often-used interpretation is that cognitive demand interferes with memory reconsolidation. Laboratory models applying this technique often do not meet some important procedural steps thought necessary to trigger reconsolidation. It remains therefore unclear whether cognitively demanding interventions can alter the reconsolidation process of aversive memories. Here, 78 (41 included) healthy participants completed an established 3-day threat conditioning paradigm. Two conditioned stimuli were paired with a shock (CS+ s) and one was not (CS-). The next day, one CS+ (CS+ R), but not the other (CS+), was presented as a reminder. After 10 min, participants performed a 2-back working memory task. On day three, we assessed retention. We found successful acquisition of conditioned threat and retention (CS+ s > CS-). However, SCRs to the CS+ R and the CS+ during retention did not significantly differ. Although threat conditioning was successful, the well-established cognitively demanding intervention did not alter the reconsolidation process of conditioned threat memories. These findings challenge current views on how cognitively demand may enhance psychotherapy-outcome.

Pattern differentiation and tuning shift in human sensory cortex underlie long-term threat memory

The amygdala-prefrontal-cortex circuit has long occupied the center of the threat system,1 but new evidence has rapidly amassed to implicate threat processing outside this canonical circuit.2-4 Through nonhuman research, the sensory cortex has emerged as a critical substrate for long-term threat memory,5-9 underpinned by sensory cortical pattern separation/completion10,11 and tuning shift.12,13 In humans, research has begun to associate the human sensory cortex with long-term threat memory,14,15 but the lack of mechanistic insights obscures a direct linkage. Toward that end, we assessed human olfactory threat conditioning and long-term (9days) threat memory, combining affective appraisal, olfactory psychophysics, and functional magnetic resonance imaging (fMRI) over a linear odor-morphing continuum (five levels of binary mixtures of the conditioned stimuli/CS+ and CS- odors). Affective ratings and olfactory perceptual discrimination confirmed (explicit) affective and perceptual learning and memory via conditioning. fMRI representational similarity analysis (RSA) and voxel-based tuning analysis further revealed associative plasticity in the human olfactory (piriform) cortex, including immediate and lasting pattern differentiation between CS and neighboring non-CS and a late onset, lasting tuning shift toward the CS. The two plastic processes were especially salient and lasting in anxious individuals, among whom they were further correlated. These findings thus support an evolutionarily conserved sensory cortical system of long-term threat representation, which can underpin threat perception and memory. Importantly, hyperfunctioning of this sensory mnemonic system of threat in anxiety further implicates a hitherto underappreciated sensory mechanism of anxiety.

Acid-sensing ion channel 1a regulates the specificity of reconsolidation of conditioned threat responses.

Recent research on altering threat memory has focused on a reconsolidation window. During reconsolidation, threat memories are retrieved and become labile. Reconsolidation of distinct threat memories is synapse dependent, whereas the underlying regulatory mechanism of the specificity of reconsolidation is poorly understood. We designed a unique behavioral paradigm in which a distinct threat memory can be retrieved through the associated conditioned stimulus. In addition, we proposed a regulatory mechanism by which the activation of acid-sensing ion channels (ASICs) strengthens the distinct memory trace associated with the memory reconsolidation to determine its specificity. The activation of ASICs by CO2 inhalation, when paired with memory retrieval, triggers the reactivation of the distinct memory trace, resulting in greater memory lability. ASICs potentiate the memory trace by altering the amygdala-dependent synaptic transmission and plasticity at selectively targeted synapses. Our results suggest that inhaling CO2 during the retrieval event increases the lability of a threat memory through a synapse-specific reconsolidation process.

Inhibiting Human Aversive Memory by Transcranial Theta-Burst Stimulation to the Primary Sensory Cortex

BackgroundPredicting adverse events from past experience is fundamental for many biological organisms. However, some individuals suffer from maladaptive memories that impair behavioral control and well-being, e.g., after psychological trauma. Inhibiting the formation and maintenance of such memories would have high clinical relevance. Previous preclinical research has focused on systemically administered pharmacological interventions, which cannot be targeted to specific neural circuits in humans. Here, we investigated the potential of noninvasive neural stimulation on the human sensory cortex in inhibiting aversive memory in a laboratory threat conditioning model. MethodsWe build on an emerging nonhuman literature suggesting that primary sensory cortices may be crucially required for threat memory formation and consolidation. Immediately before conditioning innocuous somatosensory stimuli (conditioned stimuli [CS]) to aversive electric stimulation, healthy human participants received continuous theta-burst transcranial magnetic stimulation (cTBS) to individually localized primary somatosensory cortex in either the CS-contralateral (experimental) or CS-ipsilateral (control) hemisphere. We measured fear-potentiated startle to infer threat memory retention on the next day, as well as skin conductance and pupil size during learning. ResultsAfter overnight consolidation, threat memory was attenuated in the experimental group compared with the control cTBS group. There was no evidence that this differed between simple and complex CS or that CS identification or initial learning were affected by cTBS. ConclusionsOur results suggest that cTBS to the primary sensory cortex inhibits threat memory, likely by an impact on postlearning consolidation. We propose that noninvasive targeted stimulation of the sensory cortex may provide a new avenue for interfering with aversive memories in humans.

Experiential avoidance, uncompassionate self-responding, and peritraumatic depersonalization/derealization: A novel mediation model for war-related PTSD symptomatology.

To explore a novel model for war-related posttraumatic stress disorder (PTSD) symptomatology including emotion regulation processes, namely experiential avoidance (EA) and uncompassionate self-responding (USR), mediating the impact of childhood threat memories, combat exposure distress, combat and noncombat threats, and peritraumatic depersonalization/derealization (PDD) on PTSD symptomatology. A sample of 650 male Portuguese Overseas War veterans filled self-report instruments. The model explained 59% of the variance of PTSD symptomatology. Both EA and USR mediated the effects of noncombat threats and PDD on PTSD. Additionally, EA mediated combat exposure distress and USR mediated childhood threat memories. Combat exposure distress, combat and noncombat threats, and PDD showed direct effects on PTSD symptomatology. The findings help to better understand the relationship between predictive factors of war-related PTSD in clinical and research settings, providing novel insights on the effects of combat exposure distress, and the different effects of combat and noncombat-related threats on PTSD.

An Inescapable Cat Odor Exposure Protocol for Studying Innate and Contextual Threat Conditioning in Rats.

Animals respond to threatening situations by exhibiting a number of defensive behaviors, including avoidance, freezing, and risk assessment. An animal model with an ethological approach offers a deeper insight into the biological mechanisms underlying threat responses. This paper describes a methodology to measure defensive behaviors toward both innate and learned aversive stimuli in rats. Animals were individually exposed to predator odor in an inescapable chamber to elicit a measurable, sustained, defensive state. The experimental design involved placing a rat in a familiar chamber for 10 min followed by exposure to cat odor for another 10 min in the same context. The next day, the rats were re-exposed for 10 min to the same context chamber where cat odor exposure occurred. Sessions were video-recorded and defensive behaviors were assessed on both days. The behavioral test was coupled with reversible functional inactivation and c-Fos immunohistochemistry techniques to determine the role of the interoceptive cortex in threat responses. Rats exposed to cat odor on the first day and re-exposed to the context chamber on the second day displayed higher levels of defensive behaviors, and that cat odor elicited a robust increase in the neural activity of the interoceptive cortex. Moreover, muscimol inactivation of the interoceptive cortex reduced the expression of defensive behaviors in response to cat odor and impaired contextual threat memory. These results show that this behavioral assay is a useful tool for studying neural mechanisms of defensive behaviors and may offer insight into mechanisms that mediate fear in humans and its related disorders.

A reminder before extinction failed to prevent the return of conditioned threat responses irrespective of threat memory intensity in rats.

After retrieval, reactivated memories may destabilize and require restabilization processes to persist, referred to as reconsolidation. The reminder-extinction procedure has been proposed as a behavioral reconsolidation-based intervention to persistently attenuate threat-conditioned memories. After the presentation of a single reminder trial, the conditioned threat memory may enter a labile state, and extinction training during this window can prevent the return of conditioned threat responses. However, findings on this reminder-extinction procedure are mixed and its effectiveness may be subject to boundary conditions, including memory strength. Here, we systematically investigate whether more intense threat memories are less susceptible to disruption through a reminder-extinction procedure. Using a Pavlovian auditory threat conditioning procedure at three different shock intensities, rats acquired conditioned threat responses of variable "strength." Rats subsequently underwent either extinction preceded by a reminder or standard extinction. Although different shock intensities led to different strength threat memories, all groups showed reinstatement of conditioned threat responses irrespective of shock intensity or reminder condition. Hence, regardless of the intensity of the threat memory, the reminder procedure was ineffective in preventing the return of threat responses in rats. We thus find no evidence that threat memory intensity is a potential modulator of the effectiveness of the reminder-extinction procedure. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Interactions of Noradrenergic, Glucocorticoid and Endocannabinoid Systems Intensify and Generalize Fear Memory Traces

Systemic administration of drugs that activate the noradrenergic or glucocorticoid system potentiates aversive memory consolidation and reconsolidation. The opposite happens with the stimulation of endocannabinoid signaling under certain conditions. An unbalance of these interacting neurotransmitters can lead to the formation and maintenance of traumatic memories, whose strength and specificity attributes are often maladaptive. Here we aimed to investigate whether originally low-intensity and precise contextual fear memories would turn similar to traumatic ones in rats systemically administered with adrenaline, corticosterone, and/or the cannabinoid type-1 receptor antagonist/inverse agonist AM251 during consolidation or reconsolidation. The high dose of each pharmacological agent evaluated significantly increased freezing times at test in the conditioning context one and nine days later when given alone post-acquisition or post-retrieval. Their respective low dose produced no relative changes when given separately, but co-treatment of adrenaline with corticosterone or AM251 and the three drugs combined, but not corticosterone with AM251, produced results equivalent to those mentioned initially. Neither the high nor the low dose of adrenaline, corticosterone, or AM251 altered freezing times at test in a novel, neutral context two and ten days later. In contrast, animals receiving the association of their low dose exhibited significantly higher freezing times than controls. Together, the results indicate that newly acquired and destabilized threat memory traces become more intense and generalized after a combined interference acting synergistically and mimicking that reported in patients presenting stress-related psychiatric conditions.

Observation of others’ threat reactions recovers memories previously shaped by firsthand experiences

Information about dangers can spread effectively by observation of others' threat responses. Yet, it is unclear if such observational threat information interacts with associative memories that are shaped by the individual's direct, firsthand experiences. Here, we show in humans and rats that the mere observation of a conspecific's threat reactions reinstates previously learned and extinguished threat responses in the observer. In two experiments, human participants displayed elevated physiological responses to threat-conditioned cues after observational reinstatement in a context-specific manner. The elevation of physiological responses (arousal) was further specific to the context that was observed as dangerous. An analogous experiment in rats provided converging results by demonstrating reinstatement of defensive behavior after observing another rat's threat reactions. Taken together, our findings provide cross-species evidence that observation of others' threat reactions can recover associations previously shaped by direct, firsthand aversive experiences. Our study offers a perspective on how retrieval of threat memories draws from associative mechanisms that might underlie both observations of others' and firsthand experiences.