Fear Conditioning IMPC_FEA_001

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Purpose

This procedure is used for measuring aversive learning and memory. A neutral conditioned stimulus (CS) such as steady tone is paired with an aversive unconditioned stimulus (US) such as one or more mild foot shocks. After conditioning, the spatial context or the CS (tone) elicits a central state of fear in the absence of the US (shock) that is expressed as reduced locomotor activity or total lack of movement (freezing). Immobility time is used as a measure of learning/memory performances.

Experimental Design

 

Equipment

  • Fear conditioning chambers
  • Arenas and components for cue and conditioning phases
  • Computer with recording software
  • Olfactory substance

Procedure

  1. Day 1: Conditioning
    1. Transport animals to the testing area.
    2. Set up conditioning arenas in the chambers ensuring that shocker, tone delivery and camera / beam break system are working correctly. If using video tracking equipment, ensure camera lenses are free of dust.
    3. Remove the animals from their cages and place in the arenas. Animals are conditioned using the training protocol which consists of a baseline period, audible tone, a single foot shock that co-terminates with the tone, and a period with no stimuli at the end.
    4. Remove animals from arenas and place back in home cage. Clean arenas.
  2. Day 2: Context
    1. To be performed 24 hours after conditioning.
    2. Transport animals to the testing area.
    3. Set up conditioning arenas in the chamber as on day 1 except for shocker and tone delivery system which are not used.
    4. Remove animals from their cages and place in the arena. No CS or US stimuli are presented.
    5. Remove animals from arenas and place back in home cage. Clean arenas.
  3. Day 2: Cue
    1. To be performed performed from 2 - 24 hours after the context protocol.
    2. Set up cue arenas using different floors, walls, olfactory cues and / or lighting from the conditioning context. Ensure that tone delivery and camera / beam break system are working correctly.
    3. Remove animals from their cages and place in the arena. Animals are exposed to a baseline period without any stimulus followed by the presentation of the tone.
    4. Remove animals from arenas and place back in home cage. Clean arenas.
  4. At the end of the cue protocol animals are placed back into their holding room and data is analyzed by collecting the number of freezing episodes and the total freezing/immobility time.

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