Trace Fear Conditioning: Procedure for Assessing Complex Hippocampal Function in Mice

doi:10.21769/BioProtoc.2475

. 2018 Aug 20;8(16):e2475.

doi: 10.21769/BioProtoc.2475.

Trace Fear Conditioning: Procedure for Assessing Complex Hippocampal Function in Mice

Vijendra Sharma¹, Noah Cohen¹, Rapita Sood¹, Hadile Ounallah-Saad^{2

3}, Shunit Gal-Ben-Ari², Kobi Rosenblum^{2

3}

Affiliations

¹ Department of Biochemistry, McGill University, Montréal, Québec, Canada.
² Sagol Department of Neurobiology, University of Haifa, Haifa, Israel.
³ Center for Gene Manipulation in the Brain, University of Haifa, Haifa, Israel.

PMID: 34395771
PMCID: PMC8328640
DOI: 10.21769/BioProtoc.2475

Trace Fear Conditioning: Procedure for Assessing Complex Hippocampal Function in Mice

Vijendra Sharma et al. Bio Protoc. 2018.

. 2018 Aug 20;8(16):e2475.

doi: 10.21769/BioProtoc.2475.

Authors

Vijendra Sharma¹, Noah Cohen¹, Rapita Sood¹, Hadile Ounallah-Saad^{2

3}, Shunit Gal-Ben-Ari², Kobi Rosenblum^{2

3}

Affiliations

¹ Department of Biochemistry, McGill University, Montréal, Québec, Canada.
² Sagol Department of Neurobiology, University of Haifa, Haifa, Israel.
³ Center for Gene Manipulation in the Brain, University of Haifa, Haifa, Israel.

PMID: 34395771
PMCID: PMC8328640
DOI: 10.21769/BioProtoc.2475

Abstract

The trace fear conditioning protocol is designed to measure hippocampal function in mice. The protocol includes a neutral conditioned stimulus (tone) and an aversive unconditioned stimulus (shock), separated in time by a trace interval. The trace interval between the tone and the shock critically involves the hippocampus and could be used to evaluate hippocampal-dependent learning and memory. In this protocol, we presented mice with five pairings of tone and shock separated by a 20 sec trace interval. Freezing was measured 24 h after conditioning to evaluate contextual memory by placing mice in the conditioned chamber. In addition, 48 h after conditioning, freezing was measured in a dark chamber, which served as a different context. This method enables precise detection of hippocampal-dependent learning and memory following pharmacological and genetic manipulations that impair or enhance hippocampal function.

Keywords: Contextual memory; Hippocampus function; Learning and memory deficits; Memory enhancement; Trace fear conditioning (TFC).

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Conflict of interest statement

Competing interestsThe authors declare that there are no conflicting and/or competing interests.

Figures

**Video 1.. Trace Fear Conditioning Protocol.**
This video describes the trace fear conditioning protocol to assess hippocampal function in mice (Animals were handled according to approved protocols and animal welfare regulations of the University of Haifa Institutional Ethics Committee).

**Figure 1.. The screenshots from FreezeFrame 3.0 software showing settings used in the TFC protocol.**
A. Conditioning trial; B. Contextual test; C. Tone and trace test.

**Figure 2.. Mice tested in the TFC protocol show evidence of contextual fear and freezing during tone and trace interval.**
A. Experimental design; B. TFC protocol used for the conditioning; C. Freezing response of mice during the conditioning, Base (% freezing during first 120 sec), Tone (average % freezing during the tones), Trace (average % freezing during the traces) and ITI (average % freezing during the ITI); D. Contextual memory test (% freezing during 300 sec in context); E. Tone and trace fear response, Base (% freezing during first 120 sec), Tone (average % freezing during the tones), Trace (average % freezing during the traces) and ITI (average % freezing during the ITI); F-G. The mice show normal extinction of fear response during tone and trace interval.

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References

1. Bangasser D. A., Waxler D. E., Santollo J. and Shors T. J.(2006). Trace conditioning and the hippocampus: the importance of contiguity. J Neurosci 26(34): 8702-8706. - PMC - PubMed
1. Beeman C. L., Bauer P. S., Pierson J. L. and Quinn J. J.(2013). Hippocampus and medial prefrontal cortex contributions to trace and contextual fear memory expression over time. Learn Mem 20(6): 336-343. - PubMed
1. Esclassan F., Coutureau E., Di Scala G. and Marchand A. R.(2009). A cholinergic-dependent role for the entorhinal cortex in trace fear conditioning. J Neurosci 29(25): 8087-8093. - PMC - PubMed
1. Gilmartin M. R., Kwapis J. L. and Helmstetter F. J.(2012). Trace and contextual fear conditioning are impaired following unilateral microinjection of muscimol in the ventral hippocampus or amygdala, but not the medial prefrontal cortex. Neurobiol Learn Mem 97(4): 452-464. - PMC - PubMed
1. Heise C., Taha E., Murru L., Ponzoni L., Cattaneo A., Guarnieri F. C., Montani C., Mossa A., Vezzoli E., Ippolito G., Zapata J., Barrera I., Ryazanov A. G., Cook J., Poe M., Stephen M. R., Kopanitsa M., Benfante R., Rusconi F., Braida D., Francolini M., Proud C. G., Valtorta F., Passafaro M., Sala M., Bachi A., Verpelli C., Rosenblum K. and Sala C.(2017). eEF2K/eEF2 pathway controls the excitation/inhibition balance and susceptibility to epileptic seizures. Cereb Cortex 27(3): 2226-2248. - PMC - PubMed

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[1] Bangasser D. A., Waxler D. E., Santollo J. and Shors T. J.(2006). Trace conditioning and the hippocampus: the importance of contiguity. J Neurosci 26(34): 8702-8706. - PMC - PubMed

[2] Bangasser D. A., Waxler D. E., Santollo J. and Shors T. J.(2006). Trace conditioning and the hippocampus: the importance of contiguity. J Neurosci 26(34): 8702-8706. - PMC - PubMed

[3] Beeman C. L., Bauer P. S., Pierson J. L. and Quinn J. J.(2013). Hippocampus and medial prefrontal cortex contributions to trace and contextual fear memory expression over time. Learn Mem 20(6): 336-343. - PubMed

[4] Beeman C. L., Bauer P. S., Pierson J. L. and Quinn J. J.(2013). Hippocampus and medial prefrontal cortex contributions to trace and contextual fear memory expression over time. Learn Mem 20(6): 336-343. - PubMed

[5] Esclassan F., Coutureau E., Di Scala G. and Marchand A. R.(2009). A cholinergic-dependent role for the entorhinal cortex in trace fear conditioning. J Neurosci 29(25): 8087-8093. - PMC - PubMed

[6] Esclassan F., Coutureau E., Di Scala G. and Marchand A. R.(2009). A cholinergic-dependent role for the entorhinal cortex in trace fear conditioning. J Neurosci 29(25): 8087-8093. - PMC - PubMed

[7] Gilmartin M. R., Kwapis J. L. and Helmstetter F. J.(2012). Trace and contextual fear conditioning are impaired following unilateral microinjection of muscimol in the ventral hippocampus or amygdala, but not the medial prefrontal cortex. Neurobiol Learn Mem 97(4): 452-464. - PMC - PubMed

[8] Gilmartin M. R., Kwapis J. L. and Helmstetter F. J.(2012). Trace and contextual fear conditioning are impaired following unilateral microinjection of muscimol in the ventral hippocampus or amygdala, but not the medial prefrontal cortex. Neurobiol Learn Mem 97(4): 452-464. - PMC - PubMed

[9] Heise C., Taha E., Murru L., Ponzoni L., Cattaneo A., Guarnieri F. C., Montani C., Mossa A., Vezzoli E., Ippolito G., Zapata J., Barrera I., Ryazanov A. G., Cook J., Poe M., Stephen M. R., Kopanitsa M., Benfante R., Rusconi F., Braida D., Francolini M., Proud C. G., Valtorta F., Passafaro M., Sala M., Bachi A., Verpelli C., Rosenblum K. and Sala C.(2017). eEF2K/eEF2 pathway controls the excitation/inhibition balance and susceptibility to epileptic seizures. Cereb Cortex 27(3): 2226-2248. - PMC - PubMed

[10] Heise C., Taha E., Murru L., Ponzoni L., Cattaneo A., Guarnieri F. C., Montani C., Mossa A., Vezzoli E., Ippolito G., Zapata J., Barrera I., Ryazanov A. G., Cook J., Poe M., Stephen M. R., Kopanitsa M., Benfante R., Rusconi F., Braida D., Francolini M., Proud C. G., Valtorta F., Passafaro M., Sala M., Bachi A., Verpelli C., Rosenblum K. and Sala C.(2017). eEF2K/eEF2 pathway controls the excitation/inhibition balance and susceptibility to epileptic seizures. Cereb Cortex 27(3): 2226-2248. - PMC - PubMed

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Trace Fear Conditioning: Procedure for Assessing Complex Hippocampal Function in Mice

Affiliations

Trace Fear Conditioning: Procedure for Assessing Complex Hippocampal Function in Mice

Authors

Affiliations

Abstract

Conflict of interest statement

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References

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