Unconscious Manipulation of Conceptual Representations with Decoded Neurofeedback Impacts Search Behavior.

The necessity of conscious awareness in human learning has been a long-standing topic in psychology and neuroscience. Previous research on non-conscious associative learning is limited by the low signal-to-noise ratio of the subliminal stimulus, and the evidence remains controversial, including failures to replicate. Using functional MRI decoded neurofeedback, we guided participants from both sexes to generate neural patterns akin to those observed when visually perceiving real-world entities (e.g., dogs). Importantly, participants remained unaware of the actual content represented by these patterns. We utilized an associative DecNef approach to imbue perceptual meaning (e.g., dogs) into Japanese hiragana characters that held no inherent meaning for our participants, bypassing a conscious link between the characters and the dogs concept. Despite their lack of awareness regarding the neurofeedback objective, participants successfully learned to activate the target perceptual representations in the bilateral fusiform. The behavioral significance of our training was evaluated in a visual search task. DecNef and control participants searched for dogs or scissors targets that were pre-cued by the hiragana used during DecNef training or by a control hiragana. The DecNef hiragana did not prime search for its associated target but, strikingly, participants were impaired at searching for the targeted perceptual category. Hence, conscious awareness may function to support higher-order associative learning. Meanwhile, lower-level forms of re-learning, modification, or plasticity in existing neural representations can occur unconsciously, with behavioral consequences outside the original training context. The work also provides an account of DecNef effects in terms of neural representational drift.

Functional network of contextual and temporal memory has increased amygdala centrality and connectivity with the retrosplenial cortex, thalamus, and hippocampus.

In fear conditioning with time intervals between the conditioned (CS) and unconditioned (US) stimuli, a neural representation of the CS must be maintained over time to be associated with the later US. Usually, temporal associations are studied by investigating individual brain regions. It remains unknown, however, the effect of the interval at the network level, uncovering functional connections cooperating for the CS transient memory and its fear association. We investigated the functional network supporting temporal associations using a task in which a 5-s interval separates the contextual CS from the US (CFC-5s). We quantified c-Fos expression in forty-nine brain regions of male rats following the CFC-5s training, used c-Fos correlations to generate functional networks, and analyzed them by graph theory. Control groups were trained in contextual fear conditioning, in which CS and US overlap. The CFC-5s training additionally activated subdivisions of the basolateral, lateral, and medial amygdala; prelimbic, infralimbic, perirhinal, postrhinal, and intermediate entorhinal cortices; ventral CA1 and subiculum. The CFC-5s network had increased amygdala centrality and higher amygdala internal and external connectivity with the retrosplenial cortex, thalamus, and hippocampus. Amygdala and thalamic nuclei were network hubs. Functional connectivity among these brain regions could support CS transient memories and their association.

Fear in the mind's eye: the neural correlates of differential fear acquisition to imagined conditioned stimuli.

Mental imagery is involved in both the expression and treatment of fear-related disorders such as anxiety and post-traumatic stress disorder. However, the neural correlates associated with the acquisition and generalization of differential fear conditioning to imagined conditioned stimuli are relatively unknown. In this study, healthy human participants (n = 27) acquired differential fear conditioning to imagined conditioned stimuli paired with a physical unconditioned stimulus (i.e. mild shock), as measured via self-reported fear, the skin conductance response and significant right anterior insula (aIn) activation. Multivoxel pattern analysis cross-classification also demonstrated that the pattern of activity in the right aIn during imagery acquisition was quantifiably similar to the pattern produced by standard visual acquisition. Additionally, mental imagery was associated with significant differential fear generalization. Fear conditioning acquired to imagined stimuli generalized to viewing those same stimuli as measured with self-reported fear and right aIn activity, and likewise fear conditioning to visual stimuli was associated with significant generalized differential self-reported fear and right aIn activity when imagining those stimuli. Together, the study provides a novel understanding of the neural mechanisms associated with the acquisition of differential fear conditioning to imagined stimuli and that of the relationship between imagery and emotion more generally.

Placebo effects on cutaneous pain and itch: a systematic review and meta-analysis of experimental results and methodology.

ABSTRACT: Placebo effects, positive treatment outcomes that go beyond treatment processes, can alter sensations through learning mechanisms. Understanding how methodological factors contribute to the magnitude of placebo effects will help define the mechanisms by which these effects occur. We conducted a systematic review and meta-analysis of experimental placebo studies in cutaneous pain and itch in healthy samples, focused on how differences in methodology contribute to the resulting placebo effect magnitude. We conducted meta-analyses by learning mechanism and sensation, namely, for classical conditioning with verbal suggestion, verbal suggestion alone, and observational learning, separately for pain and itch. We conducted subgroup analyses and meta-regression on the type of sensory stimuli, placebo treatment, number of acquisition and evocation trials, differences in calibrated intensities for placebo and control stimuli during acquisition, age, and sex. We replicated findings showing that a combination of classical conditioning with verbal suggestion induced larger placebo effects on pain ( k = 68, g = 0 . 59) than verbal suggestion alone ( k = 39, g = 0.38) and found a smaller effect for itch with verbal suggestion alone ( k = 7, g = 0.14). Using sham electrodes as placebo treatments corresponded with larger placebo effects on pain than when topical gels were used. Other methodological and demographic factors did not significantly affect placebo magnitudes. Placebo effects on pain and itch reliably occur in experimental settings with varied methods, and conditioning with verbal suggestion produced the strongest effects. Although methods may shape the placebo effect to some extent, these effects appear robust overall, and their underlying learning mechanisms may be harnessed for applications outside the laboratory.

Facilitated extinction but impaired extinction recall by eye movement manipulation in humans - Indications for action mechanisms and the applicability of eye movement desensitization.

Eye movement desensitization and reprocessing (EMDR) therapy utilizes the manipulation of eye movements to reduce affective distress during fear-exposure. Animal research recently suggested a potential neural mechanism underlying these effects, by which increased activity of the superior colliculus (SC), mediating visual attention, increases the inhibition of the basolateral amygdala (BLA), mediating defensive plasticity. We tested such mechanism in forty healthy humans using a multiple-day single-cue fear conditioning and extinction paradigm. The activity of the SC during extinction was experimentally manipulated by eye movements, as half of the participants executed saccadic eye movements (n = 20; major SC involvement), while the other half executed smooth eye pursuits (n = 20; minor SC involvement). Amygdala-mediated fear-potentiated startle responses and fear bradycardia, as well as threat expectancy was analyzed. Saccadic eye movements facilitated the extinction of fear bradycardia and fear-potentiated startle responses. Higher saccadic accuracy and range correlated with reduced fear-potentiated startle. However, during extinction recall, fear-potentiated startle and fear bradycardia resurged and partly reached levels obtained after fear acquisition. Threat expectancy was not affected by different eye movements and was not elevated during extinction recall. Within limitations, results support an inhibitory SC-BLA pathway in humans by which eye movements may reduce low-level defensive responding, but not threat expectancy. Yet, manipulating eye movements during extinction learning seems to impair extinction recall for behavioral and physiological defensive response indices. Thus, increasing SC activity might enhance initial efficacy of exposure treatment, but additional strategies seem necessary for sustained fear attenuation.

Thalamus sends information about arousal but not valence to the amygdala.

RATIONALE: The basolateral amygdala (BLA) and medial geniculate nucleus of the thalamus (MGN) have both been shown to be necessary for the formation of associative learning. While the role that the BLA plays in this process has long been emphasized, the MGN has been less well-studied and surrounded by debate regarding whether the relay of sensory information is active or passive. OBJECTIVES: We seek to understand the role the MGN has within the thalamoamgydala circuit in the formation of associative learning. METHODS: Here, we use optogenetics and in vivo electrophysiological recordings to dissect the MGN-BLA circuit and explore the specific subpopulations for evidence of learning and synthesis of information that could impact downstream BLA encoding. We employ various machine learning techniques to investigate function within neural subpopulations. We introduce a novel method to investigate tonic changes across trial-by-trial structure, which offers an alternative approach to traditional trial-averaging techniques. RESULTS: We find that the MGN appears to encode arousal but not valence, unlike the BLA which encodes for both. We find that the MGN and the BLA appear to react differently to expected and unexpected outcomes; the BLA biased responses toward reward prediction error and the MGN focused on anticipated punishment. We uncover evidence of tonic changes by visualizing changes across trials during inter-trial intervals (baseline epochs) for a subset of cells. CONCLUSION: We conclude that the MGN-BLA projector population acts as both filter and transferer of information by relaying information about the salience of cues to the amygdala, but these signals are not valence-specified.

Two case reports of tapering sedative-hypnotic drugs through classical conditioning using herbal medicine (CARE-compliant).

RATIONALE: Sedative-hypnotic drugs (SHDs) used for insomnia are recommended for short-term use owing to concerns regarding abuse and dependence. Nevertheless, drug discontinuation is challenging owing to rebound insomnia that occurs when the SHD is ceased. Therefore, a strategy is required to reduce or discontinue SHDs, while minimizing rebound insomnia. PATIENT CONCERNS: The present report discusses the cases of a 72-year-old man and 27-year-old man with insomnia. Both patients had been taking SHDs for more than 2 months. DIAGNOSES: Both patients were diagnosed with primary insomnia. INTERVENTIONS AND OUTCOMES: After 2 weeks of classical conditioning (CC) using Hwangryunhaedok-tang (HHT), both patients succeeded in tapering SHD; the Korean version of the insomnia severity index of patient 1 dropped from 14 to 5, and that of patient 2 dropped from 28 to 11. In the final follow-up, the index dropped to 2 and 3, respectively. LESSONS: The CC, a concept emerging in psychology, could be a new strategy for tapering drugs. In this case, SHDs, which had been taken for a long time, were successfully reduced or discontinued through CC using HHT. Herbal medicine such as HHT is selected as a neutral stimulus for CC because the risk of rebound phenomenon is low as the compounds of herbal medicine have high structural similarities with human metabolites. Both patients in this study reported no particular challenges when HHT was discontinued. However, further studies are needed that will assist in tapering anticonvulsants or steroids as well as SHDs.

Heartfulness Cleaning and Meditation: Short-Term Effects on Attention and Affect.

Heartfulness meditation (HM) trains the practitioner's attention as they progress towards reaching a super-conscious state. The process is supported by guided "Heartfulness cleaning," which helps clear the mind. This study aimed to examine the short-term effects of HM on affect and cognition and determine whether performing Heartfulness cleaning beforehand influenced the meditation outcome. Forty-eight experienced meditators (age range: 19-71 years and a male-to-female ratio: 27:21) were randomly assigned to 3 sessions: (i) HM, (ii) Heartfulness meditation preceded by cleaning, and (iii) quiet rest as a control. Mood state and emotional well-being were assessed before and after each intervention using established scales such as the Brief Mood Introspection Scale, Global Vigor and Affect Scale, Spielberger's State-Trait Anxiety Inventory, and the Digit Letter Substitution Test. After engaging in both HM and Heartfulness cleaning meditation (HCM) practices, there was a noticeable increase in feelings of pleasantness (7.3%, 7.0%, respectively) and positivity (7.5%, 7.8%, respectively), accompanied by a decrease in negative affect (14.4%, 16.5%, respectively). Additionally, HM and HCM increased in the net and total scores on a substitution test designed to measure associative learning. In contrast, there were no changes observed after 30 minutes of non-meditation. In summary, the findings of this study provide support for the positive impact of Heartfulness meditation and Heartfulness cleaning meditation on emotions, as well as their ability to enhance performance in tasks involving complex attention and associative learning. It should be noted that preceding Heartfulness meditation with 5 minutes of Heartfulness cleaning did not significantly alter the overall outcome of the meditation practice.

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.

Fear in the Theater of the Mind: Differential Fear Conditioning With Imagined Stimuli.

Many symptoms of anxiety and posttraumatic stress disorder are elicited by fearful mental imagery. Yet little is known about how visual imagery of conditioned stimuli (CSs) affects the acquisition of differential fear conditioning. Across three experiments with younger human adults (Experiment 1: n = 33, Experiment 2: n = 27, Experiment 3: n = 26), we observed that participants acquired differential fear conditioning to both viewed and imagined percepts serving as the CSs, as measured via self-reported fear and skin conductance responses. Additionally, this differential conditioning generalized across CS-percept modalities such that differential conditioning acquired in response to visual percepts generalized to the corresponding imagined percepts and vice versa. This is novel evidence that perceived and imagined stimuli engage learning processes in very similar ways and is consistent with the theory that mental imagery is depictive and recruits neural resources shared with visual perception. Our findings also provide new insight into the mechanisms of anxiety and related disorders.

Mental imagery can generate and regulate acquired differential fear conditioned reactivity.

Mental imagery is an important tool in the cognitive control of emotion. The present study tests the prediction that visual imagery can generate and regulate differential fear conditioning via the activation and prioritization of stimulus representations in early visual cortices. We combined differential fear conditioning with manipulations of viewing and imagining basic visual stimuli in humans. We discovered that mental imagery of a fear-conditioned stimulus compared to imagery of a safe conditioned stimulus generated a significantly greater conditioned response as measured by self-reported fear, the skin conductance response, and right anterior insula activity (experiment 1). Moreover, mental imagery effectively down- and up-regulated the fear conditioned responses (experiment 2). Multivariate classification using the functional magnetic resonance imaging data from retinotopically defined early visual regions revealed significant decoding of the imagined stimuli in V2 and V3 (experiment 1) but significantly reduced decoding in these regions during imagery-based regulation (experiment 2). Together, the present findings indicate that mental imagery can generate and regulate a differential fear conditioned response via mechanisms of the depictive theory of imagery and the biased-competition theory of attention. These findings also highlight the potential importance of mental imagery in the manifestation and treatment of psychological illnesses.

Acquisition and expression of fat conditioned flavor preferences following dopamine D1, opioid and NMDA receptor antagonism in C57BL/6 mice.

Objectives: Inbred mouse strains differ in the pharmacology mediating sugar and fat intake and conditioned flavor preferences (CFP). C57BL/6, BALB/c and SWR inbred mice are differentially sensitive to dopamine (DA) D1, opioid and muscarinic receptor antagonism of sucrose, saccharin or fat intake, and to DA, opioid, muscarinic and N-methyl-D-aspartate (NMDA) receptor antagonism of acquisition of sucrose-CFP. DA D1, opioid and NMDA receptor antagonists differentially alter fat (Intralipid)-CFP in BALB/c and SWR mice. The present study examined whether naltrexone, SCH23390 or MK-801 altered acquisition and expression of Intralipid-CFP in C57BL/6 mice.Methods: In acquisition, groups of male food-restricted C57BL/6 mice received vehicle, naltrexone (1, 5 mg/kg), SCH23390 (50, 200 nmol/kg) or MK-801 (100, 200 µg/kg) before 10 training sessions in which mice alternately consumed two novel-flavored 5% (CS+) and 0.5% (CS-) Intralipid solutions. Six two-bottle CS choice tests followed with both flavors mixed in 0.5% Intralipid without injections. In expression, C57BL/6 mice underwent the 10 training sessions without injections followed by two-bottle CS choice tests 30 min following vehicle, naltrexone (1, 5 mg/kg), SCH23390 (200, 800 nmol/kg) or MK-801 (100, 200 µg/kg).Results: Fat-CFP acquisition in C57BL/6 mice was significantly though marginally reduced following naltrexone, SCH23390 and MK-801. Fat-CFP expression was similarly reduced by naltrexone, SCH23390 and MK-801 in C57BL/6 mice. Discussion: C57BL/6 mice were more sensitive to DA D1, opioid and NMDA antagonists in the expression of fat-CFP relative to sugar-CFP, but were less sensitive to DA D1 and NMDA antagonists in the acquisition of fat-CFP relative to sugar-CFP.

Imaginal extinction and the vividness of mental imagery: Exploring the reduction of fear within the mind's eye.

Patients are encouraged to produce vivid mental imagery during imaginal exposure, as it is assumed to promote fear reduction. Nevertheless, the link between fear reduction and imagery vividness is unclear. We investigated the impact of vividness on fear responses using an experimental analogue of imaginal exposure - imaginal extinction - in which conditioned fear, measured with skin conductance, is reduced through exposure to mental imagery of the conditioned stimulus. We examined (1) if task-specific vividness (high vs low) of the conditioned stimulus during imaginal extinction moderated the reduction of fear responses, and (2) if task-specific vividness influenced remaining fear responses 24 h later. Findings suggest that high vividness may be advantageous for fear reduction during imaginal extinction, but it may not influence fear responses in the longer term. A possible clinical implication is that high imagery vividness during imaginal exposure may not be vital for overall treatment outcome. As high vividness is associated with increased levels of distress, a future direction would be to explore whether similar fear reduction can be obtained with less vivid imaginal exposure and thereby make treatment tolerable for more patients.

Adolescent THC Treatment Does Not Potentiate the Behavioral Effects in Adulthood of Maternal Immune Activation.

Both in utero exposure to maternal immune activation and cannabis use during adolescence have been associated with increased risk for the development of schizophrenia; however, whether these exposures exert synergistic effects on brain function is not known. In the present study, mild maternal immune activation (MIA) was elicited in mice with prenatal exposure to polyinosinic-polycytidylic acid (poly(I:C)), and ∆9-tetrahydrocannabinol (THC) was provided throughout adolescence in cereal (3 mg/kg/day for 5 days). Neither THC nor MIA pretreatments altered activity in assays used to characterize hyperdopaminergic states in adulthood: amphetamine hyperlocomotion and prepulse inhibition of the acoustic startle reflex. Adolescent THC treatment elicited deficits in spatial memory and enhanced spatial reversal learning in adult female mice in the Morris water maze, while exposure to MIA elicited female-specific deficits in fear extinction learning in adulthood. There were no effects in these assays in adult males, nor were there interactions between THC and MIA in adult females. While doses of poly(I:C) and THC were sufficient to elicit behavioral effects, particularly relating to cognitive performance in females, there was no evidence that adolescent THC exposure synergized with the risk imposed by MIA to worsen behavioral outcomes in adult mice of either sex.

Functional organization of the midbrain periaqueductal gray for regulating aversive memory formation.

Innately aversive experiences produce rapid defensive responses and powerful emotional memories. The midbrain periaqueductal gray (PAG) drives defensive behaviors through projections to brainstem motor control centers, but the PAG has also been implicated in aversive learning, receives information from aversive-signaling sensory systems and sends ascending projections to the thalamus as well as other forebrain structures which could control learning and memory. Here we sought to identify PAG subregions and cell types which instruct memory formation in response to aversive events. We found that optogenetic inhibition of neurons in the dorsolateral subregion of the PAG (dlPAG), but not the ventrolateral PAG (vlPAG), during an aversive event reduced memory formation. Furthermore, inhibition of a specific population of thalamus projecting dlPAG neurons projecting to the anterior paraventricular thalamus (aPVT) reduced aversive learning, but had no effect on the expression of previously learned defensive behaviors. By contrast, inactivation of dlPAG neurons which project to the posterior PVT (pPVT) or centromedial intralaminar thalamic nucleus (CM) had no effect on learning. These results reveal specific subregions and cell types within PAG responsible for its learning related functions.

Regulation of learned fear expression through the MgN-amygdala pathway.

Heightened fear responding is characteristic of fear- and anxiety-related disorders, including post-traumatic stress disorder. Neural plasticity in the amygdala is essential for both initial fear learning and fear expression, and strengthening of synaptic connections between the medial geniculate nucleus (MgN) and amygdala is critical for auditory fear learning. However, very little is known about what happens in the MgN-amygdala pathway during fear recall and extinction, in which conditional fear decreases with repeated presentations of the auditory stimulus alone. In the present study, we found that optogenetic inhibition of activity in the MgN-amygdala pathway during fear retrieval and extinction reduced expression of conditional fear. While this effect persisted for at least two weeks following pathway inhibition, it was specific to the context in which optogenetic inhibition occurred, linking MgN-BLA inhibition to facilitation of extinction-like processes. Reduced fear expression through inhibition of the MgN-amygdala pathway was further characterized by similar synaptic expression of GluA1 and GluA2 AMPA receptor subunits compared to what was seen in controls. Inhibition also decreased CREB phosphorylation in the amygdala, similar to what has been reported following auditory fear extinction. We then demonstrated that this effect was reduced by inhibition of GluN2B-containing NMDA receptors. These results demonstrate a new and important role for the MgN-amygdala pathway in extinction-like processes, and show that suppressing activity in this pathway results in a persistent decrease in fear behavior.

Hypericum perforatum L. prevents the acquisition of and promotes resilience against stress-induced reinstatement of the conditioned place preference induced by cocaine.

Cocaine use disorder is a serious problem worldwide, and there are no approved medications for its treatment. A novel approach to the treatment of drug addiction is the use of natural products, and, in this context, preclinical evidence suggests that Hypericum perforatum L. (Hypericum) is effective against alcohol and other substance use disorders. We hypothesised that Hypericum could also be useful as a treatment for cocaine use disorder, and so we set out to test its effectiveness in a mice model of cocaine addiction. In the first experiment we evaluated its effects on the acquisition of cocaine-induced conditioned place preference (CPP). Adult male mice were conditioned with cocaine (25 mg/kg), cocaine with Hypericum (75, 150 or 300 mg/kg) or the plant extract alone (300 mg/kg). In the second experiment, we tested the effects of Hypericum on stress-induced reinstatement of cocaine CPP. All the mice were conditioned with cocaine (25 mg/kg) and, after extinction of CPP, the reinstating effects of social defeat (alone or with 75, 150 or 300 mg/kg of Hypericum) were evaluated. All the doses of Hypericum prevented the acquisition of cocaine-induced CPP. Furthermore, the plant extract dose-dependently reduced the reinstating effects of social defeat. Therefore, Hypericum is effective in reducing the rewarding effects of cocaine and prevents the stress-induced reinstatement of cocaine CPP in mice. The mechanisms underlying these positive effects of Hypericum perforatum L. need to be determined by future research. Our results endorse Hypericum as a natural treatment for cocaine dependence.

Psychophysiological evidence for fear extinction learning via mental imagery.

Imagery-based extinction procedures have long been used in the treatments of fear-related conditions. The assumption is that imagery can substitute for the perceptual stimuli in the extinction process. Yet, experimental validations of this assumption have been limited in number and some have relied exclusively on measures of autonomic reactivity without consideration of conscious feelings of fear. The current investigation sought to assess whether imagery-based exposure could lead to extinction of conditioned fear to the corresponding perceptual stimulus. Conditioned fear responses were measured by both a physiological (i.e., skin conductance response [SCR]) and a subjective (i.e., self-reported fear) measure. Participants (N = 56) first underwent perceptual differential fear conditioning, then imagery extinction, then perceptual extinction. SCR evidence was found for successful fear conditioning, generalization of fear from viewing to imagery, and most importantly, the absence of differential fear after imagery extinction upon re-exposure to the conditioned perceptual stimulus. Self-reported fear confirmed the acquisition and generalization of fear and provided evidence of a significant reduction in differential fear conditioning across extinction. Consistent with clinical evidence of the efficacy of imagery extinction and the existing limited experimental literature, this study offers support for fear extinction to perceptual stimuli via imagery exposure.

Order does matter: the combined effects of classical conditioning and verbal suggestions on placebo hypoalgesia and nocebo hyperalgesia.

ABSTRACT: In most experimental studies in which verbal suggestion and classical conditioning are implemented together to induce placebo effects, the former precedes the latter. In naturally occurring situations, however, the information concerning pain does not always precede but often follows the pain experience. Moreover, this information is not always congruent with experience. This study investigates whether the chronology of verbal suggestion and conditioning, as well as their congruence, affects placebo hypoalgesia and nocebo hyperalgesia. The effects induced in 15 groups were compared. The participants in 8 experimental groups were presented with verbal suggestions that were either congruent or incongruent with classical conditioning. The verbal suggestions were provided either before or after conditioning. In 2 other experimental groups, placebo conditioning or nocebo conditioning was implemented without any verbal suggestion; in 2 groups, verbal suggestion of hypoalgesia or hyperalgesia without conditioning was applied. The control groups without any suggestions or conditioning were also included. Placebo hypoalgesia induced by congruent procedures was significantly stronger when the suggestion of hypoalgesia preceded rather than followed conditioning. The order of the congruent procedures did not affect the magnitude of nocebo hyperalgesia. In the groups in which incongruent procedures were implemented, placebo hypoalgesia or nocebo hyperalgesia was in line with the direction of the last-used procedure, regardless of whether it was conditioning or verbal suggestion. The results show that not the type of the procedure (verbal suggestion or conditioning), but the direction of the last-used procedure shapes pain-related expectancies and determines placebo effects.

Heterogeneous associative plasticity in the auditory cortex induced by fear learning - novel insight into the classical conditioning paradigm.

We used two-photon calcium imaging with single-cell and cell-type resolution. Fear conditioning induced heterogeneous tuning shifts at single-cell level in the auditory cortex, with shifts both to CS+ frequency and to the control CS- stimulus frequency. We thus extend the view of simple expansion of CS+ tuned regions. Instead of conventional freezing reactions only, we observe selective orienting responses towards the conditioned stimuli. The orienting responses were often followed by escape behavior.