Source Paper
Ventral pallidum GABA neuron inhibition augments context-appropriate defensive responses to learned threat cues
Ramirez EM, Martinez MX, Rokerya RK, Vera VA, Ruiz CM et al.
Neurobiol Stress • 2026
Auditory Cue Fear Conditioning
Objective: To assess fear responses to auditory cues that were previously paired with foot shock, and examine the effects of VP GABA inhibition on cue fear expression
This is a Auditory Cue Fear Conditioning protocol using Rat as the model organism. The procedure involves 4 procedural steps, 3 equipment items, 5 materials. Extracted from a 2026 paper published in Neurobiol Stress.
Model and subjects
Rat • Long Evans GAD1:Cre rats and Cre-negative wildtype littermates • Male and female • 75+ days old when experiments commenced • 85
Study window
~3 minute study window | ~48.5 hours hands-on
Core workflow
Auditory cue fear conditioning training • Wait period • Pre-test drug injection
Primary readouts
- Freezing behavior during cue presentations
- Extinction of cue responses within the session
Key equipment and reagents
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Protocol Steps
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Auditory cue fear conditioning training
Place rats in novel chamber with peppermint scent. After 3min baseline, deliver 20sec white noise cue followed immediately by foot shock. Repeat 3 times with 90sec intervals.
View evidence from paper
“rats were placed into the novel chamber, and a house light turned on for a 3min baseline period. Then, a 20sec auditory cue (continuous white noise: 80 dB) was played, which was followed immediately by either a low (0.3 mA) or high (0.75 mA) intensity foot shock, delivered for 2sec. Three cue/shock pairings were made during this training session, with 90sec elapsing between each”
Wait period
Allow 48 hours between training and testing sessions.
View evidence from paper
“48hrs later”
Pre-test drug injection
Inject rats with either vehicle or CNO 30 minutes before testing session.
View evidence from paper
“rats received either VEH or CNO injection, then they were placed into a different Med Associates chamber”
Auditory cue fear expression test
Place rats in different chamber with orange scent and distinct floor. After 3min baseline, present the shock-associated white noise cue 24 times with 90sec intervals. No shocks delivered.
View evidence from paper
“placed into a different Med Associates chamber with a distinct metal grid floor, and an orange scent (McCormick) for a 47min auditory cue fear expression session. After a 3min baseline period, the 20sec shock-associated white noise cue was played a total of 24 times, with 90sec elapsing between each. No shocks were delivered during this session”