Aug 28, 2024

Public workspaceFear Conditioning

DOI
dx.doi.org/10.17504/protocols.io.261ge56bdg47/v1
  • Sabina Marciano1,2,
  • Roberta Marongiu1,2
  • 1Department of Neurological Surgery, Weill Cornell Medical College, New York, NY 10065;
  • 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Jacquelyn Haytayan
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DOI: dx.doi.org/10.17504/protocols.io.261ge56bdg47/v1
Protocol Citation: Sabina Marciano, Roberta Marongiu 2024. Fear Conditioning. protocols.io https://dx.doi.org/10.17504/protocols.io.261ge56bdg47/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 04, 2024
Last Modified: September 23, 2024
Protocol Integer ID: 97806
Keywords: ASAPCRN, motor impairment, mouse, behavior
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: 020608
Abstract
Behavioral assay to measure fear learning and memory in mice.
Materials
  • MedAssociates fear conditioning chambers
  • white floor inserts
  • black ceiling inserts
  • pepperminet and limonene spray
Fear Acquisition
Fear Acquisition
Habituate mice to the room. Set up fear conditioning chambers as well as protocols (listed below) in VideoFreeze software.
Place individual mice in a lit fear conditioning apparatus scented with peppermint spray.
Protocol:
  • Duration00:02:00 baseline followed by 5 tone-shock pairings where the shock follows the tone
  • shock = 0.5mA, Duration00:00:01 is delivered after the end of the tone (75dB, 2kHz, Duration00:00:19 )
  • empty trace intervals between the shocks last Duration00:01:00
3m 20s
After the protocol runs for each mouse, place the mouse back in the home cage and clean the fear conditioning chamber thoroughly.
Contextual test is performed Duration24:00:00 after the fear acquisition test. Cued Test is performed at least Duration02:00:00 after the contextual test.

1d 2h
Contextual Test
Contextual Test
Goal: Measure the subjects' fear response to the environment where fear conditioning was received
24 hours after fear acquisition: Mice are placed in a lightened FC apparatus scented with peppermint spray.
Protocol: Neither cue nor shock are delivered during this test session (Duration Duration00:07:30 )

7m 30s
Cued Test
Cued Test
Goal: Measure the subjects fear response to the CS, and ideally no other features of the training environment. The key is to alter the chamber so the subjects are exposed to a new set of environmental cues.
Mice are placed in a lightened FC apparatus which is altered by placing a smooth floor insert on shock grid and black triangles on top.

Limonene spray is used, instead of peppermint scent.
Protocol:
  • Baseline: 2 min followed by five tone-cues
  • No shock is delivered after the end of the tone (75dB, 2 kHz, Duration00:00:19 )

19s
Primary Analyses
Primary Analyses
Fear conditioning acquisition
  • % Component freezing for baseline, each tone (if used), shock, and interstimulus interval
  • Graphs for each component should be graphed separately
  • Average motion
Context
  • % Component freezing per minute and total
  • Per minute and total will be on separate graphs
  • Average motion per minute and total
Cued
  • %Component freezing for baseline, each tone, and interstimulus interval
  • Graphs for each component should be graphed separately
  • Average motion for baseline, each tone, and interstimulus interval