Amphetamine Sensitization Protocol
Objective: To determine whether amphetamine sensitization enhances cue-triggered reward wanting in a drug-free state, and to assess the role of nucleus accumbens in this effect
Gather these items before starting the experiment. Check off items as you prepare.
Equipment3
Materials5
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Protocol Steps
Lever pressing training
Rats learned to lever press for sucrose pellets in operant conditioning chambers
Note: Initial training phase to establish instrumental behavior
View evidence from paper
“Rats learned to lever press for sucrose pellets”
Pavlovian cue association training
Rats separately learned to associate sucrose pellets with Pavlovian auditory cues lasting 30 seconds
Note: Establishes conditioned stimulus (CS+) association with reward
View evidence from paper
“they separately learned to associate sucrose pellets with Pavlovian cues (30 sec auditory cues)”
Amphetamine sensitization induction
Six daily injections of amphetamine (3 mg/kg, i.p.) administered to experimental group; control group received saline injections
Note: Intraperitoneal injection route; controls receive equivalent saline injections
View evidence from paper
“Amphetamine sensitization was induced by six daily injections of amphetamine (3 mg/kg, i.p.; controls received saline)”
Waiting period
Rats allowed to rest after sensitization induction before testing
Note: Drug-free period before behavioral testing
View evidence from paper
“Rats were tested for lever pressing under extinction conditions 10 d later”
Intra-accumbens microinjection
Bilateral microinjection into nucleus accumbens of either vehicle or amphetamine (5 μg/0.5 μl per side)
Note: Performed immediately before behavioral testing; counterbalanced across groups
View evidence from paper
“after a bilateral microinjection of intra-accumbens vehicle or amphetamine (5 μg/0.5 μl per side)”
Extinction testing with cue presentation
Rats tested for lever pressing under extinction conditions with intermittent presentations of free conditioned stimulus (CS+) sucrose cue
Note: Measures cue-triggered pursuit of reward; baseline pressing and CS- (negative cue) also assessed
View evidence from paper
“Rats were tested for lever pressing under extinction conditions 10 d later... Cue-triggered pursuit of sucrose reward was assessed by increases in pressing on the sucrose-associated lever during intermittent presentations of a free conditioned stimulus (CS+) sucrose cue”
Behavioral observation
Assessment of intrusive cue-triggered pursuit behaviors such as investigatory sniffing during testing
Note: Observed in sensitized rats receiving intra-accumbens amphetamine microinjection
View evidence from paper
“The amphetamine microinjection also interacted synergistically in sensitized rats to produce intrusive cue-triggered pursuit behaviors (e.g., investigatory sniffing)”