Name: A Prediction Error-driven Retrieval Procedure for Destabilizing and Rewriting Maladaptive Reward Memories in Hazardous Drinkers (Video) | JoVE Journal
Uploaded: 2018-01-05T12:30:00.000+00:00
Duration: 8 min 5 s
Description: Learn a behavioral memory retrieval procedure to destabilize maladaptive reward memories in hazardous drinkers using prediction error. This method enables testing interventions during the reconsolidation window.

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5 minutes

This work was funded in its entirety by a UK Medical Research Council grant (grant reference: MR/M007006/1) awarded to Sunjeev Kamboj and Ravi Das. The refinement of the memory retrieval procedure and sourcing of cue images was performed by Grace Gale. We would like to thank Katie Walsh and Josie Hanniford for contributions towards piloting the procedure.

All authors have no intellectual, financial, or biomedical conflicts of interest to disclose.

The protocol in this paper describes an alcohol MRM retrieval procedure that explicitly generates prediction error in alcohol delivery (Retrieval + PE) to maximize the probability that naturalistic alcohol MRMs will destabilize. This retrieval procedure takes account of recent experimental work demonstrating the necessity of prediction error for memory destabilization and engaging the reconsolidation process60. It has been shown to produce the most profound changes in indices of alcohol MRMs from subsequent behavioral interventions (counterconditioning and cognitive reappraisal) in two papers28,46 and is currently being further validated in drug models. Since this effect appears to be independent of learning history, the procedure represents a significant improvement over other memory reactivation techniques, which do not incorporate a PE, and should be used as a platform for the assessment of post-destabilization interventions for reducing maladaptive drinking behaviour. Such refinement of techniques is particularly important in light of null findings in the reconsolidation field (see below). Moreover, the RET+PE method has considerable applied significance and may be applicable in combination with a variety of post-retrieval relearning methods. For example, we have demonstrated effects on semantic memory and craving using RET+PE followed by reappraisal, a prototypical adaptive emotion regulation technique commonly used in cognitive behavioural therapy46.
The procedure is easily modified to suit the characterisitics of the participant group. For example, the stimuli used as CS+s during counterconditioning can be changed to wine pictures, if the participants are primarily wine preferring. Alternatively, smoking-related stimuli can be used if the procedure is employed in tobacco smokers.
Effective counterconditioning relies on potent disgust cues. To ensure sufficient relearning during reconsolidation, select visual UCSs that are likely to elicit powerful disgust responses. Although these are visual cues, they should have relevance to gustatory disgust (e.g., contamination-related images; images of food spoilage). If an alternative gustatory UCS (other than denatonium benzoate at the concentrations used here) is used, we recommend a compound that strongly activates bitterness responses.
Although we use an expectancy violation, it is theoretically possible to generate PE using other methods, although their efficacy in destabilizing robustly encoded reward memories has yet to be demonstrated. We have trialed an alternative procedure involving unexpected devaluation of alcohol (using unexpectedly bitter tasting alcohol), although we did not demonstrate convincing memory modifications using this procedure46.
Randomization of participants to groups should prevent baseline differences in trait questionnaire measures. If this is the case, however, these differences should be modelled as random effects in mixed-effects analyses of outcomes. A supplementary approach, which the authors recommend, is to include an extra baseline day prior to the retrieval and counterconditioning day. This allows better assessment of pre-existing group differences, the reliability of these differences, and gives researchers the option of stratified randomization to groups to prevent such differences.
In order to generate an effective PE, it is essential that participants experience a relevant surprising occurrence during retrieval. If an expectancy violation is used, as described here, ensure that participants are observed during this step so that they do not inadvertently consume the alcohol at a point they are instructed not to. Related to this, it is essential that participants are not aware of the experimenter&#39;s intention to withhold alcohol after generating the expectation of alcohol reward. As such, the protocol involves a necessary degree of deception. Upon debriefing, ensure participants are aware that they should not discuss the protocol with others.
While there is evidence that the procedure robustly destabilizes memories and has been designed to maximize the probability of doing so, there is currently no independent means of assessing whether or not destabilization has occurred. This is a limitation of all memory destabilization procedures described to date and as such, null effects of post-destabilization interventions at test are difficult to interpret, as they may be due to low efficacy of the intervention or a failure to sufficiently destabilize MRMs in the first place.
The remarkable generalization of Ret+PE dependent counterconditioning effects to non-trained stimuli makes this approach highly promising for clinical implementation. However, our demonstration of efficacy is limited to an experimental (non-clinical) setting with problem drinkers, who do not have an alcohol use disorder.
Relatedly, although the procedure is easily adapted, it is unknown whether the RET+PE procedure is similarly efficacious (in a modified form) in destabilizing MRMs in different (illicit) substance using populations. We believe it would be at least as effective, as alcohol MRMs are likely to be more overlearned, robust, and cross-contextual than MRMs for most illicit drugs. This remains to be verified experimentally however, and it is unknown how sensitive the efficacy of the procedure is to variations in number of cues presented or their exact nature. We therefore encourage experimentation with the procedure in different drug-using populations and with different retrieval cues, as well as tailoring post-destabilization interventions to population-specific reward maladaptations.
To address the measurement issues outlined above - namely the reliance on efficacy of the post-destabilization intervention to infer effective memory destabilization - independent measures of destabilization are required. We are currently developing electroencephalographic and psychophysiological techniques for resolving this issue and will make these available upon request when completed.
Implementation across the range of severities of problem drinking is required to establish the clinical utility of this technique. Moreover, clinically-oriented studies are essential to determine whether this procedure will be acceptable to treatment seekers.

Building on seminal work of Lewis and colleagues1 over the last twenty years has highlighted an unprecedented potential for plasticity in established (consolidated) memories via the process of memory reconsolidation2,3. Reconsolidation occurs under certain circumstances when memories are retrieved4 and consists of two temporally5 and molecularly6 dissociable stages: initial destabilization7&#160;and subsequent restabilization8. Following retrieval-induced memory destabilization, memories enter an &#39;active&#39;, malleable state, in which they are susceptible to modification through incorporation of novel information9,10,11,12, or weakening through pharmacological interference with the molecular pathways required for restabilization13,14,15,16. This &#39;reconsolidation window,&#39; the interval between destabilization and restabilization, lasts 1 to 6 h and represents a unique opportunity to directly interfere with the maladaptive reward memories (MRMs) that are believed to play a key role in the aetiology17, progression18, and maintenance19,20 of substance use disorders (SUDs) like alcohol use disorder.
MRMS are associations formed through Pavlovian mechanisms that link environmental stimuli or &#39;drug cues&#39; to the availability, and intoxicating and rewarding effects of drugs21. MRMs imbue drug cues with enhanced value, salience22, and motivational properties such that these cues grab attention23, trigger craving and motivate drug seeking-and-using behaviour in people with SUDs. These MRMs are therefore thought to be responsible for triggering relapse when drug cues are encountered24. The primary challenge in the long-term clinical management of SUDs is therefore to reduce the ability of MRMs to promote drug-taking, which, by extension, should reduce the incidence of relapse21. The reconsolidation window offers an exciting opportunity to achieve this aim by updating MRMs while they are unstable to a form that is less likely to contribute to relapse.
Recent research has shown that this can be achieved using purely behavioral interventions10,25. These can produce profound reductions in craving and drug-related reward metrics in heroin addicts26, smokers27, and hazardous drinkers28. By giving a brief reminder of MRMs (typically brief exposure to drug cues) to destabilize them, and subsequently re-associating drug cues with lack of drug (extinction), or aversive outcomes (counterconditioning)29 while MRMs are unstable, the associations between drug cues and outcomes restabilize in an updated, more adaptive form. These rewritten MRMs no longer support relapse. The only difference between these and standard extinction or counterconditioning interventions is the retrieval and destabilization of MRMs - produced by brief exposure to drug cues - prior to the learning intervention. They are therefore typically referred to as &#39;retrieval-extinction&#39; or &#39;retrieval-counterconditioning&#39; interventions.
However, there is now a significant body of research showing inconsistent results with these interventions30,31,32,33. This may be partially explained by individual differences in the expression of plasticity-promoting proteins34. However, since retrieval is necessary but not sufficient4,35,36 for memory destabilization, there are also certain boundary conditions that determine whether a memory destabilizes or not. The extent to which these boundary conditions were operative likely also varied from study-to-study. Chief among these conditions are memory age (chronology)37 and strength (reinforcement history)38,39, duration of the reminder trial5, and the occurrence of mismatch35,40 between predicted and actual outcome during retrieval, known as prediction error (PE)41,42,43. These factors are inter-related, as PE is the central reward &#39;learning signal&#39;44,45 and diminishes over the course of learning. PE encodes the difference between expected outcome encoded in memory and actual outcome, reflecting the inaccuracy of learned predictions. It is by minimizing the occurrence of PE that learning accrues. Old, strongly reinforced memories like MRMs - which have formed over many years - have extremely good predictive power for outcomes across different situations. As such, little PE will be generated when MRMs are retrieved under normal conditions, conferring these traces with destabilization-resistance.
Leveraging this knowledge, we developed a retrieval and PE-generation (Retrieval + PE) procedure for alcohol MRMs that aims to maximize the likelihood of these memories destabilizing by maximizing PE at retrieval. We have shown that subsequent counterconditioning causes comprehensive updating of alcohol MRMs in hazardous drinkers28,46. This manuscript describes the implementation of the Retrieval + PE procedure for destabilizing alcohol MRMs, along with a multisensory counterconditioning technique that rewrites these memories.
The Retrieval + PE procedure involves the use of verbal instructions to maximize participants&#39; expectancy of alcohol reward, then preventing alcohol consumption at the last moment. Without knowledge of the exact learning history, this PE maximization putatively maximizes the likelihood of MRMs destabilization and provides a platform for testing the efficacy of post-destabilization behavioral interventions in improving indices of maladaptive reward memory. Here we describe how to implement this procedure with subsequent disgust-based counterconditioning, which we28 and others29 have shown to be an effective learning modality for updating MRMs. However, the Retrieval + PE procedure could also be used in combination with alternative post-retrieval drug interventions and many other forms of learning-based interventions25.

<table><tbody><tr><td>Multipipette M4 x 1</td><td>Eppendorf</td><td>4982 000.012</td><td>High-accuracy pipette for dosing Bitrex</td></tr><tr><td>Combitips advanced 0.2ml x 1000</td><td>Eppendorf</td><td>0030 089.413</td><td>Syringe tip for use with Micropipette for dosing Bitrex</td></tr><tr><td>Simax 100ml measuring cylinder</td><td>Phillip Harris</td><td>B8A73325</td><td>Any suitable measuring cylinder may be used</td></tr><tr><td>Bitrex (2.5% w/v aqueaous Denatonium Benzoate solution)</td><td>Mcfarlan Smith</td><td>N/A</td><td>Bitter solution for counterconditioning drink UCSs</td></tr><tr><td>International Affective Picture System images</td><td>International Affective Picture System (Lang et al, 2008)</td><td>(http://csea.phhp.ufl.edu/media/iapsmessage.html)</td><td>Upon request from Margaret Bradley&amp;nbsp;</td></tr><tr><td>Stimulus presentation monitor with 1024 x768 colour display minimum</td><td>Dell&amp;nbsp;</td><td>N/A</td><td></td></tr><tr><td>Eyelink 1000 Core System</td><td>SR Research</td><td>Desktop-type mount</td><td>Other eye-tracking systems can be used</td></tr><tr><td>Experiment Builder</td><td>SR Resesarch</td><td>N/A</td><td>Other systems (Presentation/MATLAB/ePrime/Psychopy etc) can be used</td></tr><tr><td>DataViewer</td><td>SR Resesarch</td><td>N/A</td><td>Use software as appropriate to eye tracker setup</td></tr><tr><td>SPSS 21.0 for Windows</td><td>IBM</td><td>Base Package</td><td>Use any appopriate statistical software</td></tr><tr><td>Lion Alcometer 500</td><td>Lion Laboratories</td><td>N/A</td><td>Any accurate alcohol breath testing device may be used</td></tr><tr><td>Terumo 5ml syringe</td><td>MediSupplies</td><td>PMCO595</td><td>Any 5ml syringe may be used</td></tr><tr><td>Mini Table Top Scale</td><td>On Balance</td><td>MTT-200</td><td>Any high-accuracy scales may be used</td></tr><tr><td>Non-alcoholic beer</td><td>Bavaria</td><td>N/A</td><td>Any brand non-acloholic beer may be used</td></tr></tbody></table>

a prediction error-driven retrieval procedure for destabilizing and rewriting maladaptive reward memories in hazardous drinkers

A Prediction Error-driven Retrieval Procedure for Destabilizing and Rewriting Maladaptive Reward Memories in Hazardous Drinkers

Maladaptive reward memories (MRMs) can become unstable following retrieval under certain conditions, allowing their modification by subsequent new learning. However, robust (well-rehearsed) and chronologically old MRMs, such as those underlying substance use disorders, do not destabilize easily when retrieved. A key determinate of memory destabilization during retrieval is prediction error (PE). We describe a retrieval procedure for alcohol MRMs in hazardous drinkers that specifically aims to maximize the generation of PE and therefore the likelihood of MRM destabilization. The procedure requires explicitly generating the expectancy of alcohol consumption and then violating this expectancy (withholding alcohol) following the presentation of a brief set of prototypical alcohol cue images (retrieval + PE). Control procedures involve presenting the same cue images, but allow alcohol to be consumed, generating minimal PE (retrieval-no PE) or generate PE without retrieval of alcohol MRMs, by presenting orange juice cues (no retrieval + PE). Subsequently, we describe a multisensory disgust-based counterconditioning procedure to probe MRM destabilization by re-writing alcohol cue-reward associations prior to reconsolidation. This procedure pairs alcohol cues with images invoking pathogen disgust and an extremely bitter-tasting solution (denatonium benzoate), generating gustatory disgust. Following retrieval + PE, but not no retrieval + PE or retrieval-no PE, counterconditioning produces evidence of MRM rewriting as indexed by lasting reductions in alcohol cue valuation, attentional capture, and alcohol craving.

Compare group means at baseline for subjective measures. Groups should be equivalent on baseline intake of alcohol, disgust sensitivity/propensity, SOCRATES and NAEQ scores, and momentary craving (ACQ-NOW). If there are differences in these measures at baseline, analysis should continue using mixed-effects models and these measures should be included as random-effects59. Performance of these analyses are not described here. Assess frequency of positive alcohol breathalyzer readings in each group and report this as an outcome.
Pleasantness ratings of CS+s and CS-s did not differ at baseline (before counterconditioning), but diverged over trials of counterconditioning, with pleasantness ratings for beer CS+s reducing significantly through repeated pairing with the pictorial and bitter drink UCSs. If CS+s and CS-s differ at baseline, use a mixed-effects model with random intercepts and slopes to analyze these data. The CS-s are never paired with UCSs, so do not show a decline in pleasantness. This is represented by a CS Type (CS+ vs. CS-) x Trial (Baseline, Trial 1, 2, 3 &amp; 4) interaction if counterconditioning has been successful, as indicated in Figure 2 (representative data from Das et al.28)
Evidence of rewriting of MRM networks is provided by a generalized reduction in pleasantness ratings to alcohol stimuli, but not neutral stimuli at follow up (Day 8). In the RET+PE group, reduced pleasantness ratings were evident in response to CS+s used in the retrieval/counterconditioning task (Beer CS+s), as well as to novel beer and wine images rated for the first time at follow up. Statistically this was evident in a CS Type (Beer CS+, Neutral CS-, Novel Beer, Novel Wine) x Group ( No RET+PE, RET+PE, RET no PE) interaction. Example results showing this pattern taken from Das et al.28 are shown in Figure 3.
Examine this pattern of results in the optional attentional bias task. Attentional bias is calculated as dwell time on a target alcohol image - dwell time on its composition-matched non-target pair image. A score of 0 indicates no bias, positive score attentional bias towards alcohol stimuli, and a negative score an oculomotor aversion to alcohol pictures. Representative results taken from Das et al.28 are shown in Figure 4. It can be seen that an oculomotor aversion to alcohol images at follow up (Day 8) was produced in the RET+PE group only, indicated by a significant Group x CS Type interaction.
<img alt="Counterconditioning experiment diagram; cue image rating, prediction error, distraction task flow." class="xfigimg" src="/files/ftp_upload/56097/56097fig1.jpg" data-altupdatedat="1788250015000" data-alt="Figure 1"> 
Figure 1: A Schematic of the three retrieval groups and prediction error procedures. 
Key to abbreviations: RET + PE = Retrieval + Prediction Error; the group who retrieve alcohol memories and have expected alcohol withheld. RET + No PE = Retrieval + No Prediction Error; the group who retrieve alcohol memories and consume alcohol as expected. No RET + PE = No retrieval + Prediction Error; this group does not retrieve alcohol memories and has expected orange juice withheld. In vivo drink: A 150 mL glass of beer in RET + PE and RET + No PE or a 150 mL glass of orange juice in the no RET group. 1) In vivo drink: The drink relevant to the particpants' group is placed in front of participants and they are told that they will consume this drink according to prompts they will see on screen; after rating a series of pictures for pleasantness. 2) Cue image rating: Participants in RET + PE and RET + No PE rate four images of beer and two images of soft drinks (coffee and cola) for pleasantness. Brief in vivo exposure to beer and rating of beer images triggers retrieval of drinking memories in these groups. The No RET group rates four images of orange juice and the soft drink images for pleasantness and thus do not retrieve alcohol memories. 3) Prediction error: As all participants are told that they will drink the in vivo drink at the start of the procedure, PE is engendered by preventing participants in RET + PE and No RET + PE from consuming the drink in front of them. In RET + No PE, the drink is consumed as expected, engendering no PE. 4) Distractors: High working-memory tasks are performed (prose recall, digit span, and category and verbal fluency) to distract participants and disengage them from the retrieval task. 5) Intervention: a counterconditioning task where beer images are paired with disgust-inducing images and very bitter drinks. This procedure aims to create an association between beer images and the experience of disgust that overwrites existing associations in the RET + PE group due to destabilization of alcohol memories. <a href="https://cloudflare.jove.com/files/ftp_upload/56097/56097fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Pleasantness rating vs. trial; CS+ and CS- comparison; line graph; psychological study analysis." class="xfigimg" src="/files/ftp_upload/56097/56097fig2.jpg" data-altupdatedat="1788250015000" data-alt="Figure 2"> 
Figure 2: Mean pleasantness ratings (± SEM) of neutral and beer cues during counterconditioning. Representative changes in pleasantness ratings of CS+s (beer images paired with disgusting outcomes) and CS-s (neutral images paired with neutral outcomes) over the course of counterconditioning, taken from Das et al.28 As counterconditioning aims to invoke conditioned disgust in response to beer images, perceived pleasantness of these images should reduce during counterconditioning. Ratings do not differ at baseline, but reduce to CS+s over the course of counterconditioning. ** = CS+ &gt; CS- at p&lt; 0.001. <a href="https://cloudflare.jove.com/files/ftp_upload/56097/56097fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Bar chart of pleasantness ratings for groups: No RET+PE, RET+PE, RET no PE; emotional study." class="xfigimg" src="/files/ftp_upload/56097/56097fig3.jpg" data-altupdatedat="1788250015000" data-alt="Figure 3"> 
Figure 3: Mean pleasantness ratings (± SEM) of counterconditioned, novel alcohol cues and neutral cues at test. Representative generalized reductions in pleasantness ratings (liking) of alcohol images seven days after counterconditioning is performed following retrieval (RET) with prediction error (PE) (RET +PE) from Das et al.28 This pattern is consistent with rewriting of reward associations relating to alcohol in RET + PE. Key: Neutral CS- = coffee and cola images paired with neutral images during counterconditioning. Beer CS+ = beer images paired with disgusting outcomes during counterconditioning. Novel Beer = previously unseen beer images. Wine = previousy unseen wine images. * = significant pairwise comparisons at p&lt;0.05, Bonferroni corrected. <a href="https://cloudflare.jove.com/files/ftp_upload/56097/56097fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Attentional bias bar graph; comparison across RET+PE groups; measures in ms dwell time." class="xfigimg" src="/files/ftp_upload/56097/56097fig4.jpg" data-altupdatedat="1788250015000" data-alt="Figure 4"> 
Figure 4: Mean attentional bias to counterconditioned, novel alcohol cues and neutral cues at test. Representative generalized abolition of attentional bias to all alcohol stimuli in the group that underwent counterconditioning after RET + PE from Das et al28. Bars indicate mean attentional bias scores, calculated as the averaged difference in total time looking at a 'target image' containing visible alcohol vs. a composition-matched 'non target' imagelacking visible alcohol. Error bars are SEMs. Neutral CS- = coffee and cola images paired with neutral images during counterconditioning. Beer CS+ = beer images paired with disgusting outcomes during counterconditioning. Novel Beer = previously unseen beer images. Wine = previously unseen wine images. * = significant pairwise comparisons at p&lt;0.05, Bonferroni corrected. <a href="https://cloudflare.jove.com/files/ftp_upload/56097/56097fig4large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>TRIAL NUMBER</td>
			<td colspan="2">CS TYPE</td>
			<td colspan="2">UCS TYPE</td>
			<td colspan="2">UCS FILE NAME</td>
		</tr>
		<tr>
			<td>1</td>
			<td colspan="2">Beer CS+1</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>2</td>
			<td colspan="2">Neutral CS-1</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7020.jpg</td>
		</tr>
		<tr>
			<td>3</td>
			<td colspan="2">Beer CS+1</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>4</td>
			<td colspan="2">Neutral CS-2</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7019.jpg</td>
		</tr>
		<tr>
			<td>5</td>
			<td colspan="2">Beer CS+4</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">rotten_foot.jpg</td>
		</tr>
		<tr>
			<td>6</td>
			<td colspan="2">Beer CS+3</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">9325.jpg</td>
		</tr>
		<tr>
			<td>7</td>
			<td colspan="2">Beer CS+2</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>8</td>
			<td colspan="2">Beer CS+4</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">9325.jpg</td>
		</tr>
		<tr>
			<td>9</td>
			<td colspan="2">Beer CS+3</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">rotten_foot.jpg</td>
		</tr>
		<tr>
			<td>10</td>
			<td colspan="2">Neutral CS-1</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7020.jpg</td>
		</tr>
		<tr>
			<td>11</td>
			<td colspan="2">Beer CS+1</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>12</td>
			<td colspan="2">Neutral CS-2</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7019.jpg</td>
		</tr>
		<tr>
			<td>13</td>
			<td colspan="2">Neutral CS-1</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7019.jpg</td>
		</tr>
		<tr>
			<td>14</td>
			<td colspan="2">Beer CS+2</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>15</td>
			<td colspan="2">Beer CS+4</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">9405.jpg</td>
		</tr>
		<tr>
			<td>16</td>
			<td colspan="2">Beer CS+1</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>17</td>
			<td colspan="2">Beer CS+4</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">9301.jpg</td>
		</tr>
		<tr>
			<td>18</td>
			<td colspan="2">Neutral CS-2</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7019.jpg</td>
		</tr>
		<tr>
			<td>19</td>
			<td colspan="2">Beer CS+3</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">9405.jpg</td>
		</tr>
		<tr>
			<td>20</td>
			<td colspan="2">Beer CS+2</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>21</td>
			<td colspan="2">Beer CS+3</td>
			<td colspan="2">PICTORIAL</td>
			<td colspan="2">9301.jpg</td>
		</tr>
		<tr>
			<td>22</td>
			<td colspan="2">Neutral CS-2</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7019.jpg</td>
		</tr>
		<tr>
			<td>23</td>
			<td colspan="2">Beer CS+2</td>
			<td colspan="2">GUSTATORY</td>
			<td colspan="2">DRINK_SCREEN.PNG</td>
		</tr>
		<tr>
			<td>24</td>
			<td colspan="2">Neutral CS-1</td>
			<td colspan="2">NEUTRAL</td>
			<td colspan="2">7020.jpg</td>
		</tr>
	</tbody>
</table>
Table 1: Pseudorandomized trial order for counterconditioning task used in Das et al.28 
CS Type = Conditioned stimulus type. There are four beer images used in the counterconditioning. These are arbitrarily numbered Beer CS+ 1 to 4. Two neutral images of coffee and cola are also used and these are arbitrarily numbered Neutral CS-1 and Neutral CS-2. UCS Type = Unconditioned Stimulus type. Gustatory unconditioned stimuli are the words "DRINK NOW" appearing on-screen, and participants consuming 15 mL 0.067% Denatonium Benzoate solution. Pictorial UCSs are unpleasant/disgusting images sourced from the International Affective Picture System (IAPS) database and internet. Neutral UCSs are two images sourced from the IAPS database that are rated neutrally. UCS file name: this column gives the specific images to be displayed as outcomes on each trial. Numeric file names refer to IAPS database numbers. These images are available upon request to the IAPS database. Other images are available from the authors on request.

This manuscript describes a memory retrieval procedure for destabilizing robust reward memories and rewriting them with counterconditioning prior to their reconsolidation.

Maladaptive reward memories (MRMs) can become unstable following retrieval under certain conditions, allowing their modification by subsequent new ...

The overall goal of this behavioral memory retrieval procedure is to reactivate and destabilize maladaptive reward memories pertaining to drinking in heavy beer drinkers. The procedure aims to open the memory reconsolidation window to allow the testing of subsequent interventions. This method can help answer key questions in the memory reconsolidation field, such as what is the role of prediction error in destabilizing naturalistic alcohol memories, and what procedures following memory destabilization are most effective for targeting maladaptive reward memories.The main advantage of this technique is that it appears to allow destabilization of highly entrenched alcohol memories despite a lack of information about the learning history of those memories. This allows us to assess subsequent interventions within the reconsolidation window. This technique may allow us to develop effective reconsolidation-based therapies for alcohol use disorders, as it appears to circumvent a key obstacle in the field.In particular, it allows us to destabilize an entrenched maladaptive memory without knowledge of the learning history. Begin by using a micropipette to add 80 microliters of denatonium benzoate solution into a mixing glass. Use a graduated cylinder to carefully measure 120 milliliters of clean water into the mixing glass containing the denatonium benzoate solution.Stir well to mix, then evenly divide the 0.067%denatonium benzoate solution into eight clean, identical opaque cups. Lastly, to prepare for the participant, arrange the eight cups of bitter drink behind a screen such that they are easily reached by the experimenter but will be out of sight of the participant. For participants in the retrieval plus PE and retrieval no PE groups, pour 150 milliliters of chilled alcohol-free beer into a half-pint glass and place this on the table between the participant and the display screen.Note, for those in the no retrieval PE group, pour 150 milliliters of chilled orange juice into the glass and place this on the table in front of the participant. Then, present instructions about this stimulus-rating procedure. Show examples of the on-screen instructions that the participant will see when they are required to consume the drink in front of them.Instruct the participant to drink only when the green text drink now is presented. Then, using the software begin the reactivation task in which images are presented one at a time for 10 seconds each in pseudo-randomized order. Finally, record the participants pleasantness urge ratings in response to four beer or orange juice cues depending on the group and two neutral cues.After all images have been rated, direct the participants attention to the drink in front of them. Then present the first two screens that read pick up drink and prepare to drink, respectively, for five seconds each. Then for those in retrieval PE and no retrieval PE groups present the final screen that reads stop do not drink.Note, for those in the retrieval no PE group, present the final screen that reads drink now for five seconds and have the participant consume the beer. Then, after consuming or not consuming the drink, show the screen that reads put down the drink and press the space bar to continue. Remove the glass from the sight of the participants.Finally, have the participants rate out loud how expected or unexpected the preceding instructions were from minus five, completely unexpected, to five, completely expected, and write down the response. Immediately prior to starting the counter-conditioning task, have the participant in the no retrieval PE group rate the four beer CS for pleasantness once. Then, once the participant understands that they are required to make two ratings per trial, one for the initial picture and one for the outcome, begin the counter-conditioning task by pressing the space bar.Take the first UCS drink from behind the screen and place this in front of the participant in preparation for the drink now instructions. Present the first CS image in the sequence for six seconds. Have the participant rate how pleasant they find the image within the six seconds, from extremely unpleasant to extremely pleasant, using marked number keys on the keyboard.Then, present the drink now instructions for six seconds and have the participant pick up the cup and consume the liquid. After they have consumed the entire 15 milliliters, have the participant rate the pleasantness of the gustatory UCS outcome on the same minus five to plus five scale using the response keys. Remove the empty cup and replace with the next 15 milliliter cup of denatonium benzoate solution.Present the next CS in the sequence, a neutral image, for six seconds and have the participant rate how pleasant they find the image within this period using the response keys. Complete the remaining 22 trials. Finally, after all 24 trials have been completed, provide the participant with two squares of milk chocolate to remove the residual bitter taste of the denatonium benzoate solution.Before counter-conditioning, pleasantness ratings of beer and neutral CS images did not differ at baseline. However, they diverged over trials of counter-conditioning with pleasantness ratings for reinforced beer images reducing significantly through repeated pairing with the pictorial and bitter drink UCS's. In the retrieval prediction error group, reduced pleasantness ratings were evident in response to beer CS images used in the counter-conditioning task, as well as to novel beer and wine images rated for the first time at follow-up.Once mastered, this technique can be completed in a few minutes if it is performed properly. While attempting this procedure, it's important to remember that participants must expect to be able to drink the drink that they're given. Following this procedure, various memory-targeting methods like extinction, counter-conditioning, or drug intervention can be performed to assess their efficacy in weakening or updating maladaptive reward memories.After watching this video, you should have a good understanding of how to use a memory-retrieval and prediction error procedure to attempt to destabilize maladaptive reward memories. Don't forget that working with hazardous drinkers requires care and proper ethical approval, and this procedure should always be carried out in a setting that puts participants safety and comfort first.

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Research

JoVE Journal

Neuroscience

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Learn a behavioral memory retrieval procedure to destabilize maladaptive reward memories in hazardous drinkers using prediction error. This method enables testing interventions during the reconsolidation window.

A Prediction Error-driven Retrieval Procedure for Destabilizing and Rewriting Maladaptive Reward Memories in Hazardous Drinkers

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