Electrophysiological and behavioral analysis of lip touch as a component of the food stimulus in the snail Lymnaea

Electrophysiological and video recording methods were used to investigate the function of lip touch in feeding ingestion behavior of the pond snail Lymnaea stagnalis. Although this stimulus was used successfully as a conditioning stimulus (CS) in appetitive learning experiments, the detailed role of lip touch as a component of the sensory stimulus provided by food in unconditioned feeding behavior was never ascertained. Synaptic responses to lip touch in identified feeding motoneurons, central pattern generator interneurons, and modulatory interneurons were recorded by intracellular electrodes in a semi-intact preparation. We showed that touch evoked a complex but characteristic sequence of synaptic inputs on each neuron type. Touch never simply activated feeding cycles but provided different types of synaptic input, determined by the feeding phase in which the neuron was normally active in the rhythmic feeding cycle. The tactile stimulus evoked mainly inhibitory synaptic inputs in protraction-phase neurons and excitation in rasp-phase neurons. Swallow-phase neurons were also excited after some delay, suggesting that touch first reinforces the rasp then swallow phase. Video analysis of freely feeding animals demonstrated that during normal ingestion of a solid food flake the food is drawn across the lips throughout the rasp phase and swallow phase and therefore provides a tactile stimulus during both these retraction phases of the feeding cycle. The tactile component of the food stimulus is strongest during the rasp phase when the lips are actively pressed onto the substrate that is being moved across them by the radula. By using a semi-intact preparation we demonstrated that application of touch to the lips during the rasp phase of a sucrose-driven fictive feeding rhythm increases both the regularity and frequency of rasp-phase motoneuron firing compared with sucrose applied alone.     

Fear conditioning, preparedness, and the contingent negative variation

Psychophysiological research on preparedness has previously focused on autonomic nervous system parameters. The present study used electrocortical indices of fear conditioning. Subjects (n= 10) were tested under fear-relevant and fear-irrelevant conditions (1 week apart, order of conditions counterbalanced). Each condition comprised acquisition and extinction sessions. The contingent negative variation (CNV) was elicited in anticipation of forewarned slides (fear relevant: small animals; fear irrelevant: landscapes). In acquisition, reinforced conditioned stimulus (CS+) (but not nonreinforced conditioned stimulus [CS-]) slides were followed by white noise as an unconditioned stimulus (UCS). In extinction, the UCS was omitted. In the fear-relevant condition, CNV amplitude was significantly larger for CS+ than CS- in both acquisition and extinction. In the fear-irrelevant condition, CNV differentiation between CS+ and CS- was weak in both sessions. CNV was significantly larger in the fear-relevant than in the fear-irrelevant condition, for CS+ but not CS-. The findings are consistent with a preparedness interpretation and suggest that CNV under such circumstances may represent an automatic affective response to fear-relevant stimuli. Electrocortical measures could be particularly useful in examining information processing mechanisms in phobia and cognition-affect relationships generally.     

Prevention of inhibitory effect of dorsal raphe nucleus lesions on ovulation and LH surge by 5-HT 2A/2C receptor agonists in female rats

The lip and tentacle nerves of the pond snail, Lymnaea stagnalis, were characterized using electrophysiological techniques. When the activity of those nerves was induced in lip-tentacle preparations, aversive taste signals were transmitted through all the lip and tentacle nerves, but appetitive signals could be recorded only through the superior lip nerve. In the CNS immersed in high Mg2+ -high Ca2+ saline, electrical stimuli applied to any of the nerves failed to induce action potentials in one of the regulatory neurons (cerebral giant cell: CGC) involved in feeding responses, implying that the signals are polysynaptically transmitted to the CGC. Intracellular recordings revealed that the CGCs in semi-intact half-body preparations received both appetitive and aversive taste signals not only through the superior lip nerve but also through the median lip nerve. In addition, an osphradium was ruled out as a candidate for appetitive reception. The present results, together with our preceding data arrived at by the histochemical analyses, indicate that the appetitive taste transduction responsible for generating feeding responses is performed through the superior lip nerve with some contribution of the median lip nerve. The data showing that the CGC can receive various taste signals suggests that it may play a crucial role in feeding behavior as demonstrated in the study of conditioned taste-aversion.     

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.     

Learning- and expectation-related changes in the human brain during motor learning

We have studied a simple form of motor learning in the human brain so as to isolate activity related to motor learning and the prediction of sensory events. Whole-brain, event-related functional magnetic resonance imaging (fMRI) was used to record activity during classical discriminative delay eyeblink conditioning. Auditory conditioned stimulus (CS+) trials were presented either with a corneal airpuff unconditioned stimulus (US, paired), or without a US (unpaired). Auditory CS- trials were never reinforced with a US. Trials were presented pseudorandomly, 66 times each. The subjects gradually produced conditioned responses to CS+ trials, while increasingly differentiating between CS+ and CS- trials. The increasing difference between hemodynamic responses for unpaired CS+ and for CS- trials evolved slowly during conditioning in the ipsilateral cerebellar cortex (Crus I/Lobule HVI), contralateral motor cortex and hippocampus. To localize changes that were related to sensory prediction, we compared trials on which the expected airpuff US failed to occur (Unpaired CS+) with trials on which it occurred as expected (Paired CS+). Error-related signals in the contralateral cerebellum and somatosensory cortex were seen to increase during learning as the sensory prediction became stronger. The changes seen in the ipsilateral cerebellar cortex may be due either to the violations of sensory predictions, or to learning-related increases in the excitability of cerebellar neurons to presentations of the CS+.     

Conditioned diminution of the unconditioned skin conductance response

During Pavlovian conditioning the expression of a conditioned response typically serves as evidence that an association between a conditioned stimulus (CS) and an unconditioned stimulus (UCS) has been learned. However, learning-related changes in the unconditioned response (UCR) produced by a predictable UCS can also develop. In the present study, we investigated learning-related reductions in the magnitude of the unconditioned skin conductance response (SCR). Healthy volunteers participated in a differential conditioning study in which one tone (CS+) was paired with a loud white-noise UCS and a second tone (CS-) was presented alone. In addition, probe trials that consisted of UCS presentations paired with the CS+ (CS + UCS) and CS- (CS - UCS), as well as presentations of the UCS alone were included to assess UCR diminution. SCR and participants' expectations of UCS presentation were monitored during conditioning. Greater diminution of the UCR was observed to the UCS when it followed the CS+ compared to when it followed the CS- or was presented alone. Further, UCR amplitude showed an inverse relationship with the participants' ratings of UCS expectancy. However, conditioned UCR diminution was also observed independent of differential UCS expectancies. Our findings demonstrate conditioned diminution of the unconditioned SCR. Further, these findings suggest that although UCR amplitude is modified by conscious expectations of the UCS, conditioned diminution of the UCR can be expressed independent of learning-related changes in these expectations.     

Costs or benefits of emotional conditioning on cognitive processing?

We report on two experiments using a transfer-of-control procedure in evaluating costs or benefits of emotionally relevant stimuli on the processing of a cognitive task. In differential conditioning the impact of Pavlovian conditioning usually is examined by contrasting instrumental responses in the presence of an excitatory conditioned stimulus (CS+) and an inhibitory conditioned stimulus (CS-). We expanded this comparison by introducing a neutral third condition. This additional neutral condition served as a within subject control for evaluating whether the CS+ increased responding and/or the CS- decreased responding. Both experiments yielded the same result; the CS- slowed down cognitive processing while the CS+ had no impact. Thus, the proposed transfer-of-control procedure may serve as a reliable and valid research tool in the evaluation of motivation and emotion in humans.     

Appetitive-aversive interactions in Pavlovian fear conditioning

The existence of value coding and salience coding neurons in the mammalian brain, including in habenula and ventral tegmental area, has sparked considerable interest in the interactions that occur between Pavlovian appetitive and aversive conditioning. Here we studied these appetitive-aversive interactions at the behavioral level by assessing the learning that occurs when a Pavlovian appetitive conditioned stimulus (conditional stimulus, CS) serves as a CS for shock in Pavlovian fear conditioning. A Pavlovian appetitive CS was retarded in the rate at which it could be transformed into a fear CS (counterconditioning), but the presence of the appetitive CS augmented fear learning to a concurrently presented neutral CS (superconditioning). Retardation of fear learning was not alleviated by manipulations designed to restore the associability of the appetitive CS before fear conditioning but was alleviated by manipulations designed to increase the aversive quality of the shock unconditioned stimulus (US). These findings are consistent with opponent interactions between the appetitive and aversive motivational systems and provide a behavioral approach for assessing the neural correlates of these appetitive-aversive interactions.     

Modification of auditory and somatosensory system activity during pupillary conditioning in the paralyzed cat.

The role of sensory systems in the development of behavioral conditioned responses was investigated by recording multiple-unit activity in the auditory and somatosensory pathways during Pavlovian conditioning of the pupillary-dilation responses of paralyzed cats. Establishment of conditioned pupillary-dilation responses to a white noise CS+, pupillary discrimination between the CS+ and a tone CS-, and subsequent discrimination reversal provided the behavioral foundation for examining neural changes related to behavioral learning. Multiple-unit responses to the acoustic CS+ were significantly enhanced in the auditory cortex, cochlear nucleus, and somatic cortex, but not in the cuneate nucleus. The possibility that these effects could be due to changes in stimulus intensity at the sensory receptor, to mo-ement artifacts, or to feedback from skeletal responses were ruled out because the animals were immobilized. Nor could these neural changes be attributable to sensitization, as those brain areas which showed conditioned enhancement to the CS+ exhibited significantly larger responses to the CS+ than to the CS-. Furthermore, the changes in neural activity followed the significance of the CS; after reversal of the reinforcement contingencies, the amount of multiple-unit activity evoked by the stimuli gradually reversed too. Although the somatic cortex showed conditioning and discrimination, greater stimulus specificity was found in the auditory system. Only in the somatic cortex was there a significant increase in responses to the CS- as well as the CS4. Furthermore, both somatosensory loci exhibited enhanced responses to those tactile probes presented during the acoustic CS, suggesting a phasic increase in neural excitability to all stimuli. Analysis of the number of trials required to attain an acquisition criterion indicated that the neural changes occurred first in the auditory cortex, then the cochlear nucleus, followed in turn by the somatic cortex, and finally the cuneate nucleus. However, none of these neural changes preceded acquisition of conditioned pupillary dilations. These results suggest that sensory system changes are not essential for the initial associative process. These findings indicate that the study of autonomic conditioned responses may prove beneficial in seeking the critical neural events which underlie the initial association between two stimuli. A hypothetical model, which explains the development of pupillary and sensory system conditioned responses, was also presented.     

Fear but not awareness predicts enhanced sensory processing in fear conditioning

It is not clear whether enhanced cortical processing of reinforced stimuli as reported by neuroimaging studies is due to expectancy of an aversive event alone or to activation of the fear system. The present study investigated cortical and autonomic responses of aware participants using an instructed fear conditioning design. Steady-state visual evoked fields (ssVEF) and heart rate change were recorded to assess sensory processing and activation of the fear system by reinforced (CS+) and nonreinforced stimuli (CS-). Participants who showed heart rate acceleration demonstrated increased ssVEFs in visual and parietal cortex during CS+ in acquisition trials. Heart rate decelerators did not show enhanced cortical activation with respect to the CS+. Participants in both groups reported awareness of CS-US contingencies. Awareness of stimulus contingency in fear conditioning seems not to be sufficient to elicit enhanced visual cortical processing.     

Chemosensory conditioning of Hermissenda crassicornis

Bite-strike responses of Hermissenda crassicornis, elicited by chemosensory stimulation of the lips, were found to be modified when food extracts were paired with rotation-produced stimulation of the statocysts. Animals that received repeated pairings of an extract of 1 food (conditioned stimulus, CS) with rotation exhibited suppressed bite-strike responses to that food for up to 48 hr after training. This suppression was usually specific to the trained food and was pairing-specific as well. Discriminative conditioning was also demonstrated. Animals trained with 1 CS paired with rotation and a second CS that was unpaired (CS-) showed suppressed bite-strike responses to the first CS. The results demonstrate that Hermissenda can learn to avoid foods that reliably signal an aversive event and may allow an analysis of higher order conditioning phenomena.     

Hemispheric Asymmetry in Conditioning to Facial Emotional Expressions

In the present experiment, we report a right hemisphere advantage for autonomic conditioning to facial emotional expressions. Specifically, angry, but not happy, facial expressions showed significantly more resistance to extinction when presented initially to the right as compared to the left hemisphere. Slides of happy and angry faces were used as conditioned stimuli (CS+ and CS-) with shock as the unconditioned stimulus (UCS). Half of the subjects (n = 15) had the angry face as CS+ (and the happy face as CS-), the other half had the happy face as CS+ (and the angry face as CS-). During acquisition, the CSs were presented foveally. During extinction, using the Visual Half-Field (VHF) technique, half of the CS+ and CS- trials were randomly presented in the right visual half-field (initially to the left hemisphere), and half of the trials were presented in the left half-field (initially to the right hemisphere). Stimuli were presented for 210 ms during acquisition, and for 30 ms during extinction. Bilateral skin conductance responses (SCRs) were recorded. The results showed effects of acquisition only for the angry CS+ group. During extinction, there was a significant Conditioning X Half-field interaction which was due to greater SCRs to the CS+ angry face when it was presented in the left half-field. It is concluded that the present results reveal hemisphere asymmetry effects in facial emotional conditioning.     

Effects of classical conditioning on identification and cortical processing of speech syllables

The present study was designed to examine effects of learned motivational significance on processing of speech syllables in adults using a classical conditioning paradigm. Aversive white noise (unconditioned stimulus) was paired with two exemplars of /ba/ (conditioned stimulus, CS+) occurring near the category boundary of a 10-item /ba/-da/ continuum. Two corresponding /da/ syllables served as the CS- and indicated absence of white noise. High-density electroencephalogram (EEG) was recorded while participants passively listened to the stimuli. Prior to the EEG session and after intermittent conditioning, participants were asked to identify the syllables in a categorical perception task. Analysis of time-locked electrocortical data revealed amplitude modulations of the N2 (248-312 ms) component as a function of acquired stimulus properties (CS+ versus CS-). Over right hemisphere regions, negativity was specifically enhanced for the CS+ during intermittent conditioning. During extinction, this conditioning effect was paralleled by findings in the time-frequency domain, showing greater oscillatory activity in the gamma-band (25-40 Hz) range, 80-120 ms following onset of the CS+ compared with CS-. A similar pattern emerged in a later time segment of 400-600 ms (30-45 Hz). Aversive conditioning was not reflected in superior categorical perception performance. Our data indicate that electrocortical correlates of speech syllable perception are susceptible to changes induced by contingencies. Physiological differences were not manifest in behavioral advantage for a specific stimulus, i.e., the CS+, however. We conclude that training speech categorization by merely enhancing motivational relevance is not effective for conveying behavioral improvement.     

Effects of conditioned stimulus preexposure on human electrodermal conditioning

Two experiments investigated the effects of conditioned stimulus (CS) preexposure on Pavlovian differential conditioning and extinction of the skin conductance response. In both experiments, half the subjects were exposed to 20 presentations each of the CS+ and CS-, and the other half were exposed to control stimuli. CS duration was 8 sec. The unconditioned stimulus in Experiment 1 (N = 48) was a 1000 Hz tone of 80 dB which signalled a reaction time requirement, and in Experiment 2 (N = 48), it was a 1 sec burst of white noise at 105 dB. The results of Experiment 1 indicated that no-preexposure groups displayed more CS+/CS- differentiation than preexposure groups during acquisition and more resistance to extinction, at least for the first interval anticipatory response. In addition, the results of Experiment 2 indicated that no-preexposure groups displayed more differentiation than preexposure groups in terms of the second interval anticipatory response. These data constitute a demonstration of the latent inhibition effect with human subjects, and imply that there is an intrinsic relationship between the orienting response and the conditioning process.     

Arousal-related associative response characteristics of dorsal lateral geniculate nucleus neurons during acoustic Pavlovian fear conditioning

This research determined whether fear-conditioned, acoustic stimuli induce thalamic arousal reflected in associative responses in dorsal lateral geniculate nucleus (dLGN) neurons. Rabbits received a Pavlovian discriminative fear conditioning procedure in which one tone conditioned stimulus (CS +) was always paired with an aversive unconditioned stimulus (US) and another tone (CS-) was never paired with the US. Responses of single dLGN neurons to random CS+ and CS- presentations were then recorded. Nine of 15 recorded neurons demonstrated significantly greater firing during the CS+ versus the CS-. Their spontaneous activity demonstrated tonic firing during increased neocortical arousal and burst firing during decreased neocortical arousal. The results demonstrate that dLGN neurons show associative responses to fear-conditioned, acoustic stimuli and present a model for investigating the neural circuits by which such stimuli affect sensory processing at the thalamic level.     

Sleep Promotes Generalization of Extinction of Conditioned Fear

Study Objective:

To examine the effects of sleep on fear conditioning, extinction, extinction recall, and generalization of extinction recall in healthy humans.

Design:

During the Conditioning phase, a mild, 0.5-sec shock followed conditioned stimuli (CS+s), which consisted of 2 differently colored lamps. A third lamp color was interspersed but never reinforced (CS-). Immediately after Conditioning, one CS+ was extinguished (CS+E) by presentation without shocks (Extinction phase). The other CS+ went unextinguished (CS+U). Twelve hours later, following continuous normal daytime waking (Wake group, N = 27) or an equal interval containing a normal night''s sleep (Sleep group, N = 26), conditioned responses (CRs) to all CSs were measured (Extinction Recall phase). It was hypothesized that the Sleep versus Wake group would show greater extinction recall and/or generalization of extinction recall from the CS+E to the CS+U.

Setting:

Academic medical center.

Subjects:

Paid normal volunteers.

Measurements and Results:

Square-root transformed skin conductance response (SCR) measured conditioned responding. During Extinction Recall, the Group (Wake or Sleep) × CS+ Type (CS+E or CS+U) interaction was significant (P = 0.04). SCRs to the CS+E did not differ between groups, whereas SCRs to the CS+U were significantly smaller in the Sleep group. Additionally, SCRs were significantly larger to the CS+U than CS+E in the Wake but not the Sleep group.

Conclusions:

After sleep, extinction memory generalized from an extinguished conditioned stimulus to a similarly conditioned but unextinguished stimulus. Clinically, adequate sleep may promote generalization of extinction memory from specific stimuli treated during exposure therapy to similar stimuli later encountered in vivo.

Citation:

Pace-Schott EF; Milad MR; Orr SP; Rauch SL; Stickgold R; Pitman RK. Sleep promotes generalization of extinction of conditioned fear. SLEEP 2009;32(1):19-26.     

Electrophysiology of appetitive taste and appetitive taste conditioning in humans

The present study examines two novel aspects of appetitive processing and conditioning: the electrophysiological response to (a) the appetitive taste of a sweet fluid and (b) appetitively/taste conditioned visual stimuli. Event-related potentials (ERPs) were recorded from 32 subjects while they performed a taste conditioning task, in which two symbols were paired with a sweet or a neutral fluid. The results show a clear P1/N1/P2 complex, and a late positive potential (P3) with maxima at right fronto-central electrode sites, in response to the taste stimuli. Of these components particularly the P3 showed robust differences between the sweet and neutral taste. In addition, the electrophysiological response to the taste conditioned stimuli (CS), also showed the expected differences for both P2 and P3 at frontocentral electrodes. The present data demonstrate that the employed paradigm is a useful methodology to study the electrophysiological responses to unconditioned appetitive stimuli. Since appetitive conditioning is assumed to be disturbed in many psychopathological conditions, such as substance dependence, depression and eating disorders, this paradigm could be employed to get more insight in these conditions in humans.     

Aversive Pavlovian conditioning in psychopaths: peripheral and central correlates

Differential aversive Pavlovian conditioning with a foul odor as unconditioned stimulus (US) and neutral faces as conditioned stimuli (CS) was compared between 9 noncriminal psychopaths as defined by the Hare Psychopathy Checklist Revised and 12 healthy controls. Event-related potentials (ERP), heart rate, skin conductance response, corrugator EMG, and startle response potentiation as well as valence, arousal, and contingency of the CS were assessed. Whereas the healthy controls (HC) showed significant CS +/CS- differentiation, the psychopaths (PP) failed to exhibit a conditioned response although unconditioned responses were comparable between the groups. N100, P200, and P300 to the CSs revealed that psychopaths were not deficient in information processing and showed even better anticipatory responding than the HC group indicated by the terminal contingent negative variation (tCNV), that lacked, however, CS+ and CS- differentiation. These data indicate a deficit in association formation in psychopaths that may be related to deficient interaction of limbic-subcortical and cortical structures.     

Dorsal hippocampal involvement in appetitive trace conditioning and interval timing

Involvement of the dorsal hippocampus (DHPC) in acquisition of Pavlovian trace conditioning and interval timing was examined in an appetitive preparation in which presentations of one conditioned stimulus (CS) were immediately followed by food (delay conditioning), and presentations of another CS were followed by food 15 seconds after its termination (trace conditioning). DHPC lesions did not disrupt acquisition of trace conditioning, but they selectively affected the distribution of conditioned responding over the course of the trace CS in the early stage of acquisition. In addition, lesions disrupted accuracy of timing the conditioned response (CR) for both delay and trace CSs: The control subjects showed maximum CR at the time of food delivery, but in the DHPC-lesioned subjects, the maximum CR was at an earlier time point. This timing deficit did not seem to be due to impulsive responding or deficits in response inhibition because, when the early portion of the delay CS was interrupted shortly by an empty interval, the difference in the time of maximum responding between the lesioned and control subjects was eliminated. Thus, although the involvement of the DHPC in appetitive trace conditioning was not found when a gross measure of conditioning was employed, it was revealed when the temporal distribution of conditioned responding was examined on a moment-by-moment basis as in eyeblink trace conditioning studies.     

Orienting and Defensive Reactions to Phobic and Conditioned Fear Stimuli in Phobics and Normals

The aim of this study was to compare cardiovascular and electrodermal responses from subjects conditioned to fear-relevant stimuli with responses from phobics selected to fear the same stimuli. In the first session of the conditioning group slides of either a snake or a spider were presented. One of these served as a CS+ reinforced by an electrical shock and the other as a CS-, which was never followed by shock. In addition the conditioning group was exposed to two neutral pictures (flowers and mushrooms). The phobic group viewed the same type of stimuli and were administered some electrical shocks uncorrelated with the slides. In the second session, randomly formed halves of each group were shown (a) slides of a spider and a snake, or (b) two neutral ones. There were reliable acquisition effects in the conditioning group with the CS+ evoking larger palmar than dorsal skin conductance responses (SCRs) whereas the reverse was true for the CS-. During extinction differential responding to CS+ and CS- was maintained but palmar/dorsal differentiation disappeared. In the phobic group, feared and non-feared cues elicited differential responding with larger palmar SCRs for the feared cue only. The conditioning group failed to evidence heart rate differentiation during acquisition when all heart rate responses (HRRs) were collapsed into a single trial block (TB). When HRRs were grouped into 2 TBs, responses shifted from deceleration to CS+ on TB1 to acceleration on TB2 whereas responses to CS- were unaffected. During extinction, HR exhibited deceleration to both cues but more to CS+ than to CS-. The phobic group's heart rate accelerated to the feared cue and decelerated to the unfeared cues in both sessions. There was no difference between groups' responses to neutral cues during the second session. The results partially validate a conditioning analogue for specific phobias and the inconsistent aspects are discussed in terms of different coping strategies.