Postauricular reflexes elicited by soft acoustic clicks and loud noise probes: Reliability,prepulse facilitation,and sensitivity to picture contents

The startle blink reflex is facilitated during early picture viewing, then inhibited by attention during pleasant and aversive pictures compared to neutral pictures, and finally potentiated during aversive pictures specifically. However, it is unclear whether the postauricular reflex, which is elicited by the same loud acoustic probe as the startle blink reflex but enhanced by appetitive instead of defensive emotion, has the same pattern and time course of emotional modulation. We examined this issue in a sample of 90 undergraduates using serially presented soft acoustic clicks that elicited postauricular (but not startle blink) reflexes in addition to standard startle probes. Postauricular reflexes elicited by both clicks and probes correlated during food and nurturant contents, during which they were potentiated compared to neutral pictures, suggesting clicks effectively elicit emotionally modulated postauricular reflexes. The postauricular reflex was initially facilitated during the first 500 ms of picture processing but was larger during pleasant than neutral pictures throughout picture processing, with larger effect sizes during the latter half of picture processing. Across reflexes and eliciting stimuli, measures of emotional modulation had higher coefficient alphas than magnitudes during specific picture contents within each valence, indicating that only emotional modulation measures assess higher‐order appetitive or defensive processing.     

Pavlovian aversive and appetitive odor conditioning in humans: subjective, peripheral, and electrocortical changes

The effects of presynaptic guanosine-5'-O-(3-thio)triphosphate (GTPγS) on GABAergic inhibitory postsynaptic currents (IPSCs) were studied in cultured hippocampal neurons using whole-cell recordings. Inclusion of GTPγS (0.5–1 mM) in the presynaptic electrode reduced both the amplitude and paired-pulse depression of IPSCs, indicating that the probability of GABA-release had been reduced. Presynaptic GTPγS increased the depression of IPSCs by the GABA_B-receptor-agonist baclofen (10 μM), and the effect of baclofen was poorly reversible after washing. Stimulation of the GABAergic neuron at 80 Hz for 1 s was accompanied by tetanic depression of the IPSCs by 52±6% and was followed by post-tetanic potentiation (PTP), reaching a peak value of 71±21% and lasting about 100 s. IPSCs evoked after tetanic stimulation were depressed and PTP was absent when tetanic stimulation was applied within 3 min after starting injection of GTPγS into the presynaptic neuron. At longer times, basal release underlying a single IPSC was depressed. This affected the ratios recorded in response to tetanic stimulations such that tetanic depression was abolished, while PTP increased to 117±34%. In conclusion, GTPγS reduces the probability of GABA-release in both a use- and time-dependent manner, most likely through an inhibitory action on presynaptic Ca²⁺-influx through voltage-gated Ca²⁺ channels or an interaction with small GTP-binding proteins in the nerve terminals. Electronic Publication     

Postauricular reflex responses to pictures varying in valence and arousal

Reflexes are modulated by emotions. Much research has revealed that the startle reflexive eyeblink response is modulated by emotion, particularly in response to pictures of emotional scenes. Investigations of other reflexes are limited. Recently, research suggested that the postauricular reflex in response to a startling noise was modulated by emotion. In particular, pleasant stimuli enhanced the postauricular reflex. However, these first investigations were limited: One experiment demonstrated only a marginal difference between the pleasant stimuli and neutral stimuli and the other lacked the typical neutral scene comparison. The present experiment was designed to assess whether significant emotion versus neutral differences would occur. Results demonstrated that pleasant stimuli, regardless of arousal level, evoked larger postauricular reflex activation than neutral and unpleasant emotional stimuli.     

Depressed mood and emotional responding

The current study examined emotional responding in a sample of 57 college students who varied in depression symptom scores. Participants viewed pleasant, unpleasant and neutral emotional-eliciting pictures. Response measures included autonomic activity, subjective report of emotion, eyeblink startle reflex, and postauricular reflex. Findings indicated that greater depression symptoms predicted blunted responding with the postauricular reflex and eyeblink startle reflex measures. However, greater depression symptoms were not associated with blunted or heightened responding as indexed by the autonomic reactivity and self-report measures. Taken together, these results suggest that depression may be associated with both appetitive and defensive responding deficits, which has important implications for emotional responding. The findings underscore the importance of using multiple measures when investigating emotional responding associated with depression.     

Modeling startle eyeblink electromyogram to assess fear learning

Pavlovian fear conditioning is widely used as a laboratory model of associative learning in human and nonhuman species. In this model, an organism is trained to predict an aversive unconditioned stimulus from initially neutral events (conditioned stimuli, CS). In humans, fear memory is typically measured via conditioned autonomic responses or fear‐potentiated startle. For the latter, various analysis approaches have been developed, but a systematic comparison of competing methodologies is lacking. Here, we investigate the suitability of a model‐based approach to startle eyeblink analysis for assessment of fear memory, and compare this to extant analysis strategies. First, we build a psychophysiological model (PsPM) on a generic startle response. Then, we optimize and validate this PsPM on three independent fear‐conditioning data sets. We demonstrate that our model can robustly distinguish aversive (CS+) from nonaversive stimuli (CS‐, i.e., has high predictive validity). Importantly, our model‐based approach captures fear‐potentiated startle during fear retention as well as fear acquisition. Our results establish a PsPM‐based approach to assessment of fear‐potentiated startle, and qualify previous peak‐scoring methods. Our proposed model represents a generic startle response and can potentially be used beyond fear conditioning, for example, to quantify affective startle modulation or prepulse inhibition of the acoustic startle response.     

Postauricular and superior auricular reflex modulation during emotional pictures and sounds

The postauricular reflex is a relatively new psychophysiological measure of appetitive emotional processing during picture viewing. However, the degree to which other auricular (i.e., superior and anterior auricular) muscles might exhibit reflexive activity congruent with that found in the postauricular muscle has not been investigated, nor has the robustness of postauricular reflex modulation across stimulus modality. In this study, postauricular reflexes were the only reflexes that showed consistent emotional modulation across ears and genders. Additionally, postauricular reflexes were significantly modulated for both emotional pictures and sounds; in both cases, postauricular reflexes were greatest during pleasant stimuli.     

Measuring appetitive responding with the postauricular reflex

Recent research has provided evidence that the postauricular reflex (PAR) is potentiated while people view pleasant images relative to unpleasant and neutral images. The present study sought to extend these findings by investigating whether the PAR is more robustly modulated by the appetitive salience of the stimuli. Eyeblink startle and PAR responses of 51 participants were recorded while they viewed images that varied in valence and thematic content. Participants displayed significantly greater mean PAR magnitude during appetitive-related images (e.g., food, erotica, families) compared with non-appetitive-related pleasant images. These findings support the contention that the PAR is a useful tool for measuring appetitive responding in humans.     

The nursing hypothesis: an evolutionary account of emotional modulation of the postauricular reflex

The postauricular reflex (PAR) is anomalous because it seems to be potentiated during positive emotions and inhibited during negative states, unlike eyeblink and other components of the startle reflex. Two evolutionary explanations based on simian facial emotion expressions were tested. Reflexes were elicited while 47 young adult volunteers made lip pursing or grimacing poses and viewed neutral, intimidating, or appetitive photos. The PAR was enhanced during appetitive slides, but only as subjects carried out the lip-pursing maneuver. These results support the nursing hypothesis, which assumes that infant mammals instinctively retract their pinnae while nursing in order to comfortably position the head. Appetitive emotions prime the ear-retraction musculature, even in higher primates whose postauricular muscles are vestigial.     

Pupil dilation as an implicit measure of appetitive Pavlovian learning

Appetitive Pavlovian conditioning is a learning mechanism of fundamental biological and pathophysiological significance. Nonetheless, its exploration in humans remains sparse, which is partly attributed to the lack of an established psychophysiological parameter that aptly represents conditioned responding. This study evaluated pupil diameter and other ocular response measures (gaze dwelling time, blink duration and count) as indices of conditioning. Additionally, a learning model was used to infer participants’ learning progress on the basis of their pupil dilation. Twenty‐nine healthy volunteers completed an appetitive differential delay conditioning paradigm with a primary reward, while the ocular response measures along with other psychophysiological (heart rate, electrodermal activity, postauricular and eyeblink reflex) and behavioral (ratings, contingency awareness) parameters were obtained to examine the relation among different measures. A significantly stronger increase in pupil diameter, longer gaze duration and shorter eyeblink duration was observed in response to the reward‐predicting cue compared to the control cue. The Pearce‐Hall attention model best predicted the trial‐by‐trial pupil diameter. This conditioned response was corroborated by a pronounced heart rate deceleration to the reward‐predicting cue, while no conditioning effect was observed in the electrodermal activity or startle responses. There was no discernible correlation between the psychophysiological response measures. These results highlight the potential value of ocular response measures as sensitive indices for representing appetitive conditioning.     

Startle reflex facilitation as a function of classical eyeblink conditioning in humans

I investigated whether classical eyeblink conditioning increases the activating effect of the conditional stimulus (CS) on the relevant alpha motoneuron pool in the facial nucleus. The activating effect was measured by the reflex modification method, where startle reflexes, sharing the facial nerve motor pathway with conditional and unconditional eyeblink reflexes, were elicited in the presence of reinforced and nonreinforced CSs (CS+ and CS?). To reduce influences of conditional arousal on startle, a weak airpuff to the eye was used as unconditional stimulus and relatively short stimulus onset asynchronies (SOAs) of 300 and 600 ms were used in three startle tests presented while conditioning was in progress. Two groups (N= 72) received 100% or 0% reinforcement (Groups 100 and 0) of a tone CS+. The CS? was not reinforced. Three startle tests, consisting of acoustic startle probes presented alone and after CS+ and CS? onset, were conducted early, middle, and late in conditioning. Results showed increased startle reflex facilitation after CS+ onset in the late startle test in Group 100, indicating that startle increased as a function of classical conditioning. Although reflex facilitation at the 300-ms SOA in Group 100 is consistent with the hypothesis that CS presentations activated motoneurons in the facial nucleus, the present study cannot itself definitively support this hypothesis. Reflex facilitation at the 600-ms SOA in Group 100 could be influenced by conditional arousal.     

Generalization of appetitive conditioned responses

A stimulus (conditioned stimulus, CS) associated with an appetitive unconditioned stimulus (US) acquires positive properties and elicits appetitive conditioned responses (CR). Such associative learning has been examined extensively in animals with food as the US, and results are used to explain psychopathologies (e.g., substance‐related disorders or obesity). Human studies on appetitive conditioning exist, too, but we still know little about generalization processes. Understanding these processes may explain why stimuli not associated with a drug, for instance, can elicit craving. Forty‐seven hungry participants underwent an appetitive conditioning protocol during which one of two circles with different diameters (CS+) became associated with an appetitive US (chocolate or salty pretzel, according to participants’ preference) but never the other circle (CS?). During generalization, US were delivered twice and the two CS were presented again plus four circles (generalization stimuli, GS) with gradually increasing diameters from CS? to CS+. We found successful appetitive conditioning as reflected in appetitive subjective ratings (positive valence, higher contingency) and physiological responses (startle attenuation and larger skin conductance responses) to CS+ versus CS?, and, importantly, both measures confirmed generalization as indicated by generalization gradients. Small changes in CS‐US contingency during generalization may have weakened generalization processes on the physiological level. Considering that appetitive conditioned responses can be generalized to non‐US‐associated stimuli, a next important step would be to investigate risk factors that mediate overgeneralization.     

Postauricular and eyeblink startle responses to facial expressions

Emotional facial expressions have affective significance. Smiles, for example, are perceived as positive and responded to with increased happiness, whereas angry expressions are perceived as negative and threatening. Yet, these perceptions are modulated in part by facial morphological cues related to the sex of the expresser. The present research assessed both eyeblink startle and the postauricular reflex during happy and angry expressions by men and women. For this 14 male and 16 female undergraduates saw happy, neutral, and angry facial expressions as well as positive and negative pictures. The postauricular reflex was potentiated during happy expressions and inhibited during anger expressions; however, as expected, this pattern was more clearly found for female expressers. Conversely, the expected pattern of eyeblink startle potentiation during angry faces and inhibition during happy faces was found only for male expressers.     

Fear-potentiated startle conditioning in humans: Explicit and contextual cue conditioning following paired versus unpaired training

Conditioned fear in response to explicit and contextual cues was examined using the startle reflex in three groups of participants over two sessions separated by 4–5 days. The conditioned stimulus (CS) was paired with an aversive unconditioned stimulus (US) (shock) during conditioning in the paired but not in the unpaired group. In the reaction time (RT) group, the US was a nonaversive visual signal for an RT task. In the paired group, the CS potentiated startle in the postconditioning phase. This conditioned response was fully retained over the retention interval. There was no substantial change in baseline startle (startle delivered in the absence of CS). By contrast, startle was not potentiated by the CS in the unpaired group, but baseline startle was increased from Session 1 to Session 2. In the RT group, startle was not affected by the CS, and baseline startle was reduced from Session 1 to Session 2. These results suggest that paired presentations of a CS and an aversive US result in conditioned fear in response to the CS but little contextual fear, whereas unpaired presentations of a CS and US leads to poor explicit cue conditioning but substantial contextual fear.     

Affective startle modulation in anticipation and perception

Startle modulation was investigated as participants first anticipated and then viewed affective pictures in order to determine whether affective modulation of the startle reflex is similar in these different task contexts. During a 6-s anticipation period, a neutral light cue signaled whether the upcoming picture would portray snakes, erotica, or household objects; at the end of the anticipatory period, a picture in the signaled category was viewed for 6 s. Male participants highly fearful of snakes were recruited to maximize emotional arousal during anticipation and perception. Results indicated that the startle reflex was potentiated when anticipating either unpleasant (phobic) or pleasant (erotic) pictures, compared to neutral stimuli, whereas during perception, reflexes were potentiated when viewing unpleasant stimuli, and reduced when viewing pleasant pictures. The startle reflex is modulated by hedonic valence in picture perception, and by emotional arousal in a task context involving picture anticipation.     

The psychophysiology of social anxiety: Emotional modulation of the startle reflex during socially-relevant and -irrelevant pictures

The present study examined affective processes of social anxiety (SA) through emotional modulation of the startle reflex. Eighty-four high and low trait socially anxious undergraduates viewed socially-relevant and socially-irrelevant pleasant, neutral, and unpleasant pictures, and acoustic startle probes were presented during pictures and the inter-trial interval. Startle was potentiated during unpleasant compared to pleasant stimuli, but this valence modulation did not reliably vary between groups or socially-relevant and -irrelevant stimuli. However, when participants were categorized based on public-speaking fears rather than general SA symptoms, the high fear group demonstrated reliable valence modulation, whereas the low fear group did not. These findings are interpreted within the context of the broader literature suggesting that the specificity of fear in SA may influence psychophysiological reactivity.     

Affect, attention, or anticipatory arousal? Human blink startle modulation in forward and backward affective conditioning

Affect modulates the blink startle reflex in the picture-viewing paradigm, however, the process responsible for reflex modulation during conditional stimuli (CSs) that have acquired valence through affective conditioning remains unclear. In Experiment 1, neutral shapes (CSs) and valenced or neutral pictures (USs) were paired in a forward (CS-->US) manner. Pleasantness ratings supported affective learning of positive and negative valence. Post-acquisition, blink reflexes were larger during the pleasant and unpleasant CSs than during the neutral CS. Rather than affect, attention or anticipatory arousal were suggested as sources of startle modulation. Experiment 2 confirmed that affective learning in the picture-picture paradigm was not affected by whether the CS preceded the US. Pleasantness ratings and affective priming revealed similar extents of affective learning following forward, backward or simultaneous pairings of CSs and USs. Experiment 3 utilized a backward conditioning procedure (US-->CS) to minimize effects of US anticipation. Again, blink reflexes were larger during CSs paired with valenced USs regardless of US valence implicating attention rather than anticipatory arousal or affect as the process modulating startle in this paradigm.     

Chemosensory anxiety signals augment the startle reflex in humans

The aim of the present study was to investigate whether chemosensory anxiety signals can activate behavioral withdrawal systems in humans. Twelve male university students donated their axillary sweat in two situations: right before an oral academic examination (anxiety condition) and during ergometric training (exercise condition). Subjective ratings revealed that the odor donors experienced significantly more anxiety and less pleasure during the anxiety condition than during the exercise condition. Seven subjects (three females) participated in the psychophysiological experiment. The chemosensory stimuli from pooled sweat samples of the donors, and from unused cotton pads (pad condition) were presented via a constant-flow olfactometer. Acoustic startle probes (100 dB (A)) were delivered during and between the presentations of the chemosensory stimuli. Only three subjects were able to discriminate the chemosensory stimuli of the human sweat samples from room air. However, the startle reflex amplitude (EMG of the eyeblink response) recorded in the context of chemosensory anxiety signals was increased, as compared to the amplitude recorded in the context of chemosensory stimuli from either exercise (p = 0.018) or cotton pad (p = 0.012). It is concluded that chemosensory anxiety signals may pre-attentively prime defensive behavior.     

Does emotion modulate the blink reflex in human conditioning? Startle potentiation during pleasant and unpleasant cues in the picture–picture paradigm

Emotional processes modulate the size of the eyeblink startle reflex in a picture-viewing paradigm, but it is unclear whether emotional processes are responsible for blink modulation in human conditioning. Experiment 1 involved an aversive differential conditioning phase followed by an extinction phase in which acoustic startle probes were presented during CS+, CS-, and intertrial intervals. Valence ratings and affective priming showed the CS+ was unpleasant postacquisition. Blink startle magnitude was larger during CS+ than during CS-. Experiment 2 used the same design in two groups trained with pleasant or unpleasant pictorial USs. Ratings and affective priming indicated that the CS+ had become pleasant or unpleasant in the respective group. Regardless of CS valence, blink startle was larger during CS+ than CS- in both groups. Thus, startle was not modulated by CS valence.     

Startle modulation and explicit valence evaluations dissociate during backward fear conditioning

Blink startle magnitude is linearly modulated by affect such that, relative to neutral stimuli, startle magnitude is inhibited during pleasant stimuli and potentiated during unpleasant stimuli. Andreatta, Mühlberger, Yarali, Gerber, and Pauli (2010), however, report a dissociation between startle modulation and explicit valence evaluations during backward conditioning, a procedure in which the unconditional stimulus precedes the conditional stimulus (CS). Relative to controls, startles elicited during the CS were inhibited, suggesting that the CS had acquired positive valence, but participants still evaluated the CS as unpleasant after the experiment. In Experiment 1, we aimed to replicate this dissociation using a trial‐by‐trial measure of CS valence to measure startle modulation and CS valence simultaneously during forward and backward differential fear conditioning. In Experiment 2, we examined whether early and late portions of the CS could acquire differential valence by presenting startle probes at early and late probe positions during the CS. In both experiments, the dissociation between startle modulation and explicit valence evaluations in backward conditioning replicated, with CS+ evaluated as less pleasant than CS‐, but startles elicited during CS+ inhibited relative to CS‐. In Experiment 2, we provide preliminary evidence that this inhibition was present early, but not late, during the CS+. The results replicate the dissociation between implicit and explicit CS valence reported by Andreatta et al. (2010) using a trial‐by‐trial measure of valence. We also provide preliminary evidence that this dissociation may occur because the implicit and explicit measures are recorded at different times during the CS presentation.     

The development of fear learning and generalization in 8-13 year-olds

The current study examined developmental changes in fear learning and generalization in 40 healthy 8–13 year‐olds using an aversive conditioning paradigm adapted from Lau et al. [Lau et al. [2008] Journal of the American Academy of Child and Adolescent Psychiatry 47:94–102]. In this task, the conditioned stimuli (CS+/CS?) are two neutral female faces, and the unconditioned stimulus is a fearful, screaming face. The second phase of the study also included a generalization stimulus (GS): a 50% blend of the CS± faces. The eye‐blink startle reflex was utilized to measure defensive responding. Patterns of fear learning and generalization were qualified by child age. Older children demonstrated greater fear learning (i.e., larger startle during CS+ than CS?) than younger children. In addition, older children exhibited the typical pattern of generalization observed in adults, whereas younger children did not. Finally, fear learning also related to contingency awareness; only children who correctly identified the CS+ demonstrated fear‐potentiated startle to the CS+. Clinical implications and future directions are discussed. © 2011 Wiley Periodicals,Inc. Dev Psychobiol 54: 675–684, 2012.