Source Paper
Auditory Thalamus, Dorsal Hippocampus, Basolateral Amygdala, and Perirhinal Cortex Role in the Consolidation of Conditioned Freezing to Context and to Acoustic Conditioned Stimulus in the Rat
Benedetto Sacchetti, Carlo Ambrogi Lorenzini, Elisabetta Baldi, Giovanna Tassoni, Corrado Bucherelli
Journal of Neuroscience • 1999
Freezing Duration Measurement
Objective: Measure context and conditioned stimulus freezing durations during retention testing at 48 and 72 hours after tetrodotoxin administration to assess brain structure involvement in fear memory consolidation
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Protocol Steps
Fear conditioning training
Adult male Wistar rats undergo conditioned stimulus (CS) and context fear training prior to tetrodotoxin administration
Note: This is the training phase that precedes TTX injection
View evidence from paper
“fully reversible tetrodotoxin (TTX) inactivation was performed on adult male Wistar rats having undergone CS and context fear training”
Anesthesia and stereotaxic injection
Anesthetized animals receive stereotaxic injection of tetrodotoxin at increasing post-acquisition delays into target brain structures (auditory thalamus, dorsal hippocampus, basolateral amygdala, or perirhinal cortex)
Note: TTX dose and volume adjusted according to site dimensions
View evidence from paper
“Anesthetized animals were injected stereotaxically with TTX (either 5 or 10 ng in 0.5 or 1.0 µl of saline, according to site dimensions) at increasing post-acquisition delays”
Retention testing at 48 hours
Measure context and conditioned stimulus freezing durations during retention testing 48 hours after tetrodotoxin administration
Note: First retention test timepoint
View evidence from paper
“Context and CS freezing durations were measured during retention testing, always performed 48 and 72 hr after TTX administration”
Retention testing at 72 hours
Measure context and conditioned stimulus freezing durations during retention testing 72 hours after tetrodotoxin administration
Note: Second retention test timepoint
View evidence from paper
“Context and CS freezing durations were measured during retention testing, always performed 48 and 72 hr after TTX administration”