Inhibitory Avoidance Conditioning
Objective: Examine the involvement of cannabinoid CB1 receptor in the basolateral amygdala in inhibitory avoidance conditioning and extinction, and test whether cannabinoid activation reverses stress effects on these memory processes
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Equipment2
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Protocol Steps
Microinjection of WIN55,212-2 into basolateral amygdala
Microinject WIN55,212-2 into the basolateral amygdala prior to stress exposure or behavioral testing
Note: Dose: 5 μg/0.5 μl. This treatment had no effect on IA conditioning or extinction by itself
View evidence from paper
“The synthetic full agonist of the CB1/CB2 receptor WIN55,212-2 (5 μg/0.5 μl) microinjected into the BLA had no effect on IA conditioning or extinction by itself”
Stress exposure
Expose rats to a stressor following WIN55,212-2 microinjection
Note: WIN55,212-2 microinjected before stressor exposure reversed the enhancing effects of stress on IA conditioning
View evidence from paper
“microinjecting WIN55,212-2 into the BLA before exposing the rats to a stressor reversed the enhancing effects of the stressor on IA conditioning”
Inhibitory avoidance conditioning
Conduct inhibitory avoidance conditioning task to measure acquisition of inhibitory avoidance memory in response to aversive stimulus
Note: WIN55,212-2 reversed stress-induced enhancement of IA conditioning
View evidence from paper
“inhibitory avoidance (IA) conditioning and extinction and to test whether cannabinoid activation would reverse the effects of stress on these memory processes”
Inhibitory avoidance extinction
Conduct extinction trials for inhibitory avoidance memory
Note: WIN55,212-2 reversed stress-induced impairment of IA extinction. AM251 blocked extinction when microinjected into BLA
View evidence from paper
“its impairing effects on IA extinction. Importantly, WIN55,212-2 microinjected into the BLA reduced stress-induced elevations in corticosterone levels”
Microinjection of AM251 into basolateral amygdala
Microinject CB1 receptor antagonist AM251 into the basolateral amygdala to test CB1 receptor involvement in extinction
Note: Dose: 6 ng/0.5 μl. AM251 significantly blocked extinction of IA
View evidence from paper
“the CB1 receptor antagonist AM251 (6 ng/0.5 μl) microinjected into the BLA significantly blocked extinction”
Sensorimotor control testing
Test sensorimotor parameters to ensure WIN55,212-2 effects are not due to sensorimotor deficits
Note: Sensorimotor parameters remained unchanged by WIN55,212-2 microinjection into BLA
View evidence from paper
“the effects of WIN55,212-2 could not be attributed to sensorimotor deficits, because these parameters seemed unchanged by WIN55,212-2 microinjected into the BLA”
Corticosterone level measurement
Measure corticosterone levels to assess stress-induced hormonal changes
Note: WIN55,212-2 reduced stress-induced elevations in corticosterone levels
View evidence from paper
“WIN55,212-2 microinjected into the BLA reduced stress-induced elevations in corticosterone levels”