Conditioned Fear Behavior Test
Objective: Assessment of fear-induced behaviors including freezing/crouching in rats exposed to a context previously paired with footshock, and mapping neural substrates of conditioned fear through c-fos expression analysis
Gather these items before starting the experiment. Check off items as you prepare.
Equipment2
Materials1
Not specified • Not specified • Not specified • Not specified
Software1
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Protocol Steps
Conditioning procedure
Rats received footshock in a specific environment to establish classical conditioning
Note: This establishes the aversively conditioned contextual cues
View evidence from paper
“the environment in which they had previously received footshock”
Test session - Exposure to conditioned context
Rats were exposed to the environment in which they had previously received footshock
Note: Behavioral analysis was performed during this exposure to assess fear-induced behaviors
View evidence from paper
“Exposure to the environment in which they had previously received footshock significantly increased the number of Fos-like immunoreactive neurons”
Behavioral observation
Analysis of behaviors emitted by rats during test session, specifically measuring frequency of crouching (freezing) behavior
Note: Provided strong evidence that conditioning procedure was effective
View evidence from paper
“Analysis of the behaviors emitted by the rats during the test session provided strong evidence that the conditioning procedure was effective”
Diazepam pretreatment experiment
In a second experiment, rats received pretreatment with diazepam at doses of 2.5, 5.0, or 10 mg/kg prior to exposure to conditioned context
Note: Evaluated effects of anxiolytic drug on fear-induced behaviors and c-fos expression
View evidence from paper
“In a second experiment, the effects of pretreatment with the anxiolytic drug diazepam (2.5, 5.0, or 10 mg/kg) were evaluated”
Behavioral measurement with diazepam
Measurement of crouching (freezing) frequency in diazepam-pretreated rats exposed to conditioned context
Note: Benzodiazepine produced dose-related decreases in freezing behavior
View evidence from paper
“The benzodiazepine produced dose-related decreases in the frequency of crouching (freezing) elicited by the aversively conditioned contextual cues”
Immunohistochemistry and neural mapping
Analysis of Fos-like immunoreactive neurons in brain tissue to map neural substrates of conditioned fear and effects of diazepam
Note: Examined approximately 60 brain structures including cortical and subcortical regions
View evidence from paper
“The synthesis of Fos, the protein product of the immediate early gene c-fos, was used to map metabolically some of the neural substrates of conditioned fear in the rat”