An Animal Model of Genetic Vulnerability to Behavioral Disinhibition and Responsiveness to Reward-Related Cues: Implications for Addiction methods
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rat
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Response to quinpirole
reagent used in the protocol.
- Use
- Rats from the S17 generation received three doses of quinpirole (counterbalanced; 0.1, 0.3, and 1.0 mg/kg, i.p.) or three injections of vehicle (0.9% NaCL, i.p.). Behavior was videotaped during habituation and the post-injection period, and Clever Sys (Reston, VA) Drug Scan software was used to analyze video-r...
Dopamine regulation in bHR and bLR rats
reagent used in the protocol.
- Use
- Dopamine D2 receptor binding: Dopamine D2 receptor binding was measured by the competition of dopamine with [ 3 H]domperidone ( ). Brains were collected from bHRs and bLRs from the S17 generation under basal conditions, and the dorsal striatum (caudate-putamen) was dissected and rapidly frozen. This tissue was later...
Cocaine US
reagent used in the protocol.
- Use
- shows the effect of pairing presentation of a lever-CS with an intravenous injection of cocaine (US) in bHR ( n =8) and bLR ( n =8) rats. With training, bHR rats developed a sign-tracking CR to the cocaine-paired CS, whereas bLR rats did not. It is important to note that with a drug US there is no measurable goal-tr...
Response to Quinpirole
reagent used in the protocol.
- Use
- The frequency of head movements was determined by dividing the total number of lateral head movements by the time spent in place, providing an index of the vigor of drug-induced head movements when the animals are not engaged in other competing behaviors, such as locomotion (; ). There was no significant effect of...
Impulsivity
reagent used in the protocol.
- Use
- It has been suggested that robust sign-tracking behavior may reflect a lack of inhibitory control over behavior (; ), which is one attribute of impulsivity. It is interesting, therefore, to compare differences in the selected lines in sign-tracking behavior with differences on various tests of 'impulsivity'....
Impulsivity
reagent used in the protocol.
- Use
- When 'impulsive action' was assessed using a DRL task, bHRs were more impulsive-they were less able to withhold responding to receive reward. These latter results are consistent with those recently reported in outbred HR/LR rats ( ). However, another recent report using outbred rats and a different measu...
Dopamine
reagent used in the protocol.
- Use
- In summary, bHRs exhibit a number of characteristics related to addiction vulnerability and 'externalizing disorders' in humans. We had previously reported their increased propensity to risk-taking behavior (; ) and to drug self-administration ( ). In this study we show that relative to bLR rats, bHR rats als...
MATERIALS AND METHODS
Each generation of rats was screened for locomotor activity around 60 days of age (see; ). Novelty-induced locomotor data were also obtained for bHRs and bLRs from the S17 generation using an automated behavioral analysis system (CleverSys, Reston, VA; ).
- Use
- Each generation of rats was screened for locomotor activity around 60 days of age (see; ). Novelty-induced locomotor data were also obtained for bHRs and bLRs from the S17 generation using an automated behavioral analysis system (CleverSys, Reston, VA; ).
Pavlovian conditional approach
Food-unconditional stimulus: Goal- and cue-directed Pavlovian approach responses were assessed in rats from generations S13-S16 using equipment and procedures described previously (, ). Each Pavlovian training session consisted of 25 trials, in which an illuminated lever (conditional stimulus, CS) was i...
- Use
- Food-unconditional stimulus: Goal- and cue-directed Pavlovian approach responses were assessed in rats from generations S13-S16 using equipment and procedures described previously (, ). Each Pavlovian training session consisted of 25 trials, in which an illuminated lever (conditional stimulus, CS) was i...
Response to quinpirole
Rats from the S17 generation received three doses of quinpirole (counterbalanced; 0.1, 0.3, and 1.0 mg/kg, i.p.) or three injections of vehicle (0.9% NaCL, i.p.). Behavior was videotaped during habituation and the post-injection period, and Clever Sys (Reston, VA) Drug Scan software was used to analyze video-r...
- Use
- Rats from the S17 generation received three doses of quinpirole (counterbalanced; 0.1, 0.3, and 1.0 mg/kg, i.p.) or three injections of vehicle (0.9% NaCL, i.p.). Behavior was videotaped during habituation and the post-injection period, and Clever Sys (Reston, VA) Drug Scan software was used to analyze video-r...
Pavlovian Conditional Approach
The effect of Pavlovian training using food as the US (ie, pairing a lever-CS with food delivery) was examined in four independent experiments using bHR and bLR rats from generations S13-S16. The results were qualitatively and statistically similar across generations, and therefore, for the sake of simplicity,...
- Use
- The effect of Pavlovian training using food as the US (ie, pairing a lever-CS with food delivery) was examined in four independent experiments using bHR and bLR rats from generations S13-S16. The results were qualitatively and statistically similar across generations, and therefore, for the sake of simplicity,...
Pavlovian Conditional Approach
Taken together, these data indicate that bHR rats are primarily 'sign-trackers' ( ) because their CR consists of approach toward and engagement with the cue, or 'sign', that predicts reward delivery. On the other hand, bLR rats are primarily 'goal trackers' ( ) because upon CS presentation their CR...
- Use
- Taken together, these data indicate that bHR rats are primarily 'sign-trackers' ( ) because their CR consists of approach toward and engagement with the cue, or 'sign', that predicts reward delivery. On the other hand, bLR rats are primarily 'goal trackers' ( ) because upon CS presentation their CR...
Cocaine US
shows the effect of pairing presentation of a lever-CS with an intravenous injection of cocaine (US) in bHR ( n =8) and bLR ( n =8) rats. With training, bHR rats developed a sign-tracking CR to the cocaine-paired CS, whereas bLR rats did not. It is important to note that with a drug US there is no measurable goal-tr...
- Use
- shows the effect of pairing presentation of a lever-CS with an intravenous injection of cocaine (US) in bHR ( n =8) and bLR ( n =8) rats. With training, bHR rats developed a sign-tracking CR to the cocaine-paired CS, whereas bLR rats did not. It is important to note that with a drug US there is no measurable goal-tr...
Impulsivity
It has been suggested that robust sign-tracking behavior may reflect a lack of inhibitory control over behavior (; ), which is one attribute of impulsivity. It is interesting, therefore, to compare differences in the selected lines in sign-tracking behavior with differences on various tests of 'impulsivity'....
- Use
- It has been suggested that robust sign-tracking behavior may reflect a lack of inhibitory control over behavior (; ), which is one attribute of impulsivity. It is interesting, therefore, to compare differences in the selected lines in sign-tracking behavior with differences on various tests of 'impulsivity'....
Impulsivity
The fact that the different measures of impulsivity used here were dissociable is consistent with a number of other studies on impulsive action vs impulsive choice (;;; ). It is not clear why manipulations (selective breeding in this case) that increase impulsive action may sometimes also decrease impulsive choic...
- Use
- The fact that the different measures of impulsivity used here were dissociable is consistent with a number of other studies on impulsive action vs impulsive choice (;;; ). It is not clear why manipulations (selective breeding in this case) that increase impulsive action may sometimes also decrease impulsive choic...
Phasic dopamine activity
Software used for acquisition, scoring, statistics, or reporting.
- Use
- Fast-scan cyclic voltammetry was used to evaluate spontaneous and food-evoked phasic dopamine release in the core of the nucleus accumbens of bHR ( n =5) and bLR ( n =5) rats. The core of the nucleus accumbens was targeted because it has been implicated in a number of the behaviors reported here (for review see ). T...
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MATERIALS AND METHODS
A total of 260 male Sprague-Dawley rats from our selective breeding colony ( ) were used. Procedures were approved by the University Committee on the Use and Care of Animals. Unless otherwise indicated, rats were pair-housed and kept on a 12 h light/12 h dark cycle (lights on 0600 hours) with controlled temperature and humidity. Food and water were available ad libitum.
MATERIALS AND METHODS
Each generation of rats was screened for locomotor activity around 60 days of age (see; ). Novelty-induced locomotor data were also obtained for bHRs and bLRs from the S17 generation using an automated behavioral analysis system (CleverSys, Reston, VA; ).
General Statistics
Linear mixed-effects models ( ) were used to analyze the behavioral data. The covariance structure was explored and modeled appropriately for each dependent variable and when significant main effects or interactions were detected, Bonferroni post hoc comparisons were carried out. Independent t -tests were used to examine phenotypic differences in acquisition of behavior for the delay-discounting task and for the neurobiological data.
Impulsive behavior
DRL: The same rats used for the delay-discounting task were also used for the DRL task (adapted from. Test chambers were arranged such that there was just one lever located to either the right or left (counterbalanced) of the food cup and the lever remained extended for the duration of the sessions. Each 45 min session began with illumination of the house light and lever presentation. Rats were initially tested on a DRL-5-s schedule, in which a lever press resulted in illumination of the lever and pellet delivery only if at least 5 s had elapsed since the previous press. Following 5 days of training at DRL-5-s, the schedule was changed to DRL-10-s for 5 days, then to DRL-20-s for 5 days, and the last 5 days at DRL-30-s. The total number of lever presses and the total number of reinforcers (ie, successful lever presses) were recorded.
Response to quinpirole
Rats from the S17 generation received three doses of quinpirole (counterbalanced; 0.1, 0.3, and 1.0 mg/kg, i.p.) or three injections of vehicle (0.9% NaCL, i.p.). Behavior was videotaped during habituation and the post-injection period, and Clever Sys (Reston, VA) Drug Scan software was used to analyze video-recorded behavior (; ).
Cocaine US
shows the effect of pairing presentation of a lever-CS with an intravenous injection of cocaine (US) in bHR ( n =8) and bLR ( n =8) rats. With training, bHR rats developed a sign-tracking CR to the cocaine-paired CS, whereas bLR rats did not. It is important to note that with a drug US there is no measurable goal-tracking response-that is, there is no specific location associated with delivery of the cocaine. Moreover, when cocaine is used as the US, animals rarely contact the lever-CS (as they do with a food US). Instead, the sign-tracking CR consists of orientation to the lever followed by approach and exploration (often sniffing) in the immediate vicinity of the lever, similar to that described when rewarding electrical brain stimulation is used as the US (see; ). Thus, CS-directed approach behavior is illustrated in as the probability that a rat approached the lever during...
Response to Quinpirole
The behavioral response to quinpirole, a direct agonist at D2/D3 dopamine receptors ( ), was assessed in bHR ( n =10-12/group) and bLR ( n =12/group) rats. Initial analysis of the time course of the behavioral effects indicated that the peak response fell between 25 and 55 min following injection, and therefore statistical analyses were conducted using data from this 30-min period. The effect of a saline injection was assessed in an independent group of animals and therefore these data were not included in the dose-effect analyses. However, independent t -tests were performed to determine whether the response at a specific dose significantly differed from that of the saline controls.
Response to Quinpirole
illustrates the dose-response functions for locomotor distance and the frequency of head movements following quinpirole administration. There was a significant effect of phenotype for quinpirole-induced locomotor activity (F (1,23) =9.99; P =0.01) and this effect was most pronounced at the lowest dose examined ( ). bHR rats exhibited a large increase in locomotor activity in response to 0.1 mg/kg of quinpirole relative to bLR rats and it was only at this lowest dose that bHR rats significantly differed from bHR saline control rats ( P =0.005). bLRs did not differ significantly from their saline control counterparts at any dose, but there was a trend level effect at 0.3 mg/kg ( P =0.09). There was also a trend for a phenotype × dose interaction (F (2,39) =2.67, P =0.08), but no overall effect of dose. These results were likely affected by the large amount of vari...
Measurement outputs
What raw and processed outputs should exist?
Each generation of rats was screened for locomotor activity around 60 days of age (see; ). Novelty-induced locomotor data were also obtained for bHRs and bLRs from the S17 gene...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Linear mixed-effects models ( ) were used to analyze the behavioral data. The covariance structure was explored and modeled appropriately for each dependent variable and when si...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Food-unconditional stimulus: Goal- and cue-directed Pavlovian approach responses were assessed in rats from generations S13-S16 using equipment and procedures descri...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Cocaine-unconditional stimulus: The propensity to approach a cocaine-paired cue was examined using rats from the S13 generation. After catheters were implanted (; ), Pavl...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
How should the outputs become interpretable results?
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
Linear mixed-effects models ( ) were used to analyze the behavioral data.
from paperScoring or quantification
Quantify the primary readouts for this experiment: Each generation of rats was screened for locomotor activity around 60 days of age (see; ). Novelty-induced locomotor data were also obtained for bHRs and bLRs from the S17 gene...; Linear mixed-effects models ( ) were used to analyze the behavioral data. The covariance structure was explored and modeled appropriately for each dependent variable and when si...; Food-unconditional stimulus: Goal- and cue-directed Pavlovian approach responses were assessed in rats from generations S13-S16 using equipment and procedures descri...; Cocaine-unconditional stimulus: The propensity to approach a cocaine-paired cue was examined using rats from the S13 generation. After catheters were implanted (; ), Pavl....
from paperStatistical comparison
Linear mixed-effects models ( ) were used to analyze the behavioral data. The covariance structure was explored and modeled appropriately for each dependent variable and when si...; The effect of Pavlovian training using food as the US (ie, pairing a lever-CS with food delivery) was examined in four independent experiments using bHR and bLR rats from genera...; To better assess the rate of learning in bHR and bLR rats, we directly compared each of the three measures using analyses of variance, in which session was treated as a continuo...; Finally, in one additional analysis we compared the difference in the probability of approaching the lever-CS vs the food cup during the CS period across training sessions (ie,...
from paperReporting output
Report representative outputs alongside summary comparisons for Each generation of rats was screened for locomotor activity around 60 days of age (see; ). Novelty-induced locomotor data were also obtained for bHRs and bLRs from the S17 gene..., Linear mixed-effects models ( ) were used to analyze the behavioral data. The covariance structure was explored and modeled appropriately for each dependent variable and when si..., Food-unconditional stimulus: Goal- and cue-directed Pavlovian approach responses were assessed in rats from generations S13-S16 using equipment and procedures descri..., Cocaine-unconditional stimulus: The propensity to approach a cocaine-paired cue was examined using rats from the S13 generation. After catheters were implanted (; ), Pavl....
inferred from protocolStructured statistical methods
Linear mixed-effects models ( ) were used to analyze the behavioral data. The covariance structure was explored and modeled appropriately for each dependent variable and when si...; The effect of Pavlovian training using food as the US (ie, pairing a lever-CS with food delivery) was examined in four independent experiments using bHR and bLR rats from genera...; To better assess the rate of learning in bHR and bLR rats, we directly compared each of the three measures using analyses of variance, in which session was treated as a continuo...; Finally, in one additional analysis we compared the difference in the probability of approaching the lever-CS vs the food cup during the CS period across training sessions (ie,...
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Evidence quotes (8)
A total of 260 male Sprague-Dawley rats from our selective breeding colony ( ) were used. Procedures were approved by the University Committee on the Use and Care of Animals. Unless otherwise indicated, rats were pair-housed and kept on a 12 h light/12 h dark cycle (lights on 0600 hours) with controlled temperature and humidity. Food and water were available ad libitum.
Each generation of rats was screened for locomotor activity around 60 days of age (see; ). Novelty-induced locomotor data were also obtained for bHRs and bLRs from the S17 generation using an automated behavioral analysis system (CleverSys, Reston, VA; ).
Linear mixed-effects models ( ) were used to analyze the behavioral data. The covariance structure was explored and modeled appropriately for each dependent variable and when significant main effects or interactions were detected, Bonferroni post hoc comparisons were carried out. Independent t -tests were used to examine phenotypic differences in acquisition of behavior for the delay-discounting task and for the neurobiological data.
DRL: The same rats used for the delay-discounting task were also used for the DRL task (adapted from. Test chambers were arranged such that there was just one lever located to either the right or left (counterbalanced) of the food cup and the lever remained extended for the duration of the sessions. Each 45 min session began with illumination of the house light and lever presentation. Rats were initially tested on a DRL-5-s schedule, in which a lever press resulted in illumination of the lever and pellet delivery only if at least 5 s had elapsed since the previous press. Following 5 days of training at DRL-5-s, the schedule was changed to DRL-10-s for 5 days, then to DRL-20-s for 5 days, and the last 5 days at DRL-30-s. The total number of lever presses and the total number of reinforcers (ie, successful lever presses) were recorded.
Rats from the S17 generation received three doses of quinpirole (counterbalanced; 0.1, 0.3, and 1.0 mg/kg, i.p.) or three injections of vehicle (0.9% NaCL, i.p.). Behavior was videotaped during habituation and the post-injection period, and Clever Sys (Reston, VA) Drug Scan software was used to analyze video-recorded behavior (; ).
shows the effect of pairing presentation of a lever-CS with an intravenous injection of cocaine (US) in bHR ( n =8) and bLR ( n =8) rats. With training, bHR rats developed a sign-tracking CR to the cocaine-paired CS, whereas bLR rats did not. It is important to note that with a drug US there is no measurable goal-tracking response-that is, there is no specific location associated with delivery of the cocaine. Moreover, when cocaine is used as the US, animals rarely contact the lever-CS (as they do with a food US). Instead, the sign-tracking CR consists of orientation to the lever followed by approach and exploration (often sniffing) in the immediate vicinity of the lever, similar to that described when rewarding electrical brain stimulation is used as the US (see; ). Thus, CS-directed approach behavior is illustrated in as the probability that a rat approached the lever during the CS period in a given trial block (, the number of approaches the rat made toward the lever ( ) and the latency with which the rat approached the lever ( ). For detailed statistics on these measures, see Supplementary Table 2. Relative to bLRs, with training bHRs increased their probability of a...
The behavioral response to quinpirole, a direct agonist at D2/D3 dopamine receptors ( ), was assessed in bHR ( n =10-12/group) and bLR ( n =12/group) rats. Initial analysis of the time course of the behavioral effects indicated that the peak response fell between 25 and 55 min following injection, and therefore statistical analyses were conducted using data from this 30-min period. The effect of a saline injection was assessed in an independent group of animals and therefore these data were not included in the dose-effect analyses. However, independent t -tests were performed to determine whether the response at a specific dose significantly differed from that of the saline controls.
illustrates the dose-response functions for locomotor distance and the frequency of head movements following quinpirole administration. There was a significant effect of phenotype for quinpirole-induced locomotor activity (F (1,23) =9.99; P =0.01) and this effect was most pronounced at the lowest dose examined ( ). bHR rats exhibited a large increase in locomotor activity in response to 0.1 mg/kg of quinpirole relative to bLR rats and it was only at this lowest dose that bHR rats significantly differed from bHR saline control rats ( P =0.005). bLRs did not differ significantly from their saline control counterparts at any dose, but there was a trend level effect at 0.3 mg/kg ( P =0.09). There was also a trend for a phenotype × dose interaction (F (2,39) =2.67, P =0.08), but no overall effect of dose. These results were likely affected by the large amount of variance for the bHRs at 0.3 mg/kg, which was largely due to one animal that showed an unusually large locomotor response. Nevertheless, these data show that bHRs are more sensitive to a low dose of quinpirole than bLRs.
Machine-readable layer
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"text": "DRL: The same rats used for the delay-discounting task were also used for the DRL task (adapted from. Test chambers were arranged such that there was just one lever located to either the right or left (counterbalanced) of the food cup and the lever remained extended for the duration of the sessions. Each 45 min session began with illumination of the house light and lever presentation. Rats were initially tested on a DRL-5-s schedule, in which a lever press resulted in illumination of the lever and pellet delivery only if at least 5 s had elapsed since the previous press. Following 5 days of training at DRL-5-s, the schedule was changed to DRL-10-s for 5 days, then to DRL-20-s for 5 days, and the last 5 days at DRL-30-s. The total number of lever presses and the total number of reinforcers (ie, successful lever presses) were recorded."
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"name": "An Animal Model of Genetic Vulnerability to Behavioral Disinhibition and Responsiveness to Reward-Related Cues: Implications for Addiction methods",
"item": "https://replicatescience.com/experiments/an-animal-model-of-genetic-vulnerability-to-behavioral-disinhibition-and-responsiveness-to-reward-related-cues-implications-for-addiction-methods-shelly-b-flagel-pmc2794950/an-animal-model-of-genetic-vulnerability-to-behavioral-disinhibition-and-responsiveness-to-reward-re-mlph2tei"
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