Glucocorticoid receptor-mediated amygdalar metaplasticity underlies adaptive modulation of fear memory by stress
- University of Toyama, Japan
- Cairo University, Egypt
- Jichi Medical University, Japan
- The University of Tokyo, Japan
Figures
Figure 1 with 1 supplement
Generation and characterization of Grp-iCre mice.
(A) Schematic diagram of the gene targeting strategy. iCre and pgk-neo cassettes flanked by two FRT sites were inserted into the gastrin-releasing peptide gene (Grp) locus. Met is the translation …
Figure 1—figure supplement 1
Generation of floxed GR mice.
(A) Strategy for targeting the GR (Nr3c1) gene locus. The coding regions of GR exons are indicated by closed boxes. The construct included two loxP sites and the pgk-neo genes flanked by two FRT …
Figure 2 with 1 supplement
Generation and characterization of lateral amygdala (LA)-selective glucocorticoid receptor (GR) knockout (LAGRKO) mice.
(A) The LAGRKO mouse line (GRflox/flox, Grp-iCre+/-) was established by crossing floxed GR (GRflox/flox) and Grp-iCre mice. Double immunofluorescence staining of GR (green, left panels) and NeuN …
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Figure 2—source data 1
Raw data for generating Figure 2B.
- https://doi.org/10.7554/eLife.34135.006
Figure 2—figure supplement 1
Expression of GR in the LA and cerebral cortex of floxed GR and LAGRKO mice.
(A) Double immunofluorescence staining of GR (green, left panels) and NeuN (magenta, middle panels) in the LA. The overlap of green and magenta signals (white, right panels) indicates the expression …
Figure 3 with 2 supplements
Genetic disruption of LAGR impairs auditory-cued fear memory following strengthened conditioning.
(A) Experimental protocol for fear conditioning. Mice were trained with a protocol using six CS × US pairings. Contextual and auditory-cued fear memory were tested 24 and 48 hr after training, …
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Figure 3—source data 1
Raw data for generating Figure 3B–E.
- https://doi.org/10.7554/eLife.34135.010
Figure 3—figure supplement 1
LAGRKO mice did not exhibit fear memory deficits with the moderate fear conditioning protocol.
(A) Experimental design for fear conditioning. Mice were trained with three CS × US pairing protocol. Contextual and auditory fear memories were tested 24 and 48 hr after training, respectively. …
Figure 3—figure supplement 2
LAGRKO mice exhibited no changes in locomotor activity and anxiety level as determined using the open field test.
Distance of travel throughout the apparatus (A) and in the center zone of the open field (B) and time spent in the center of the open field (C) did not differ between the two genotypes. Floxed GR, n …
Figure 4
Rescue of the auditory-cued fear memory deficit in LAGRKO mice by restoring GR expression levels in the LA.
(A) The experimental protocol for adeno-associated virus (AAV) vector injection and fear conditioning. (B) Representative images of the expression of green fluorescent protein (GFP, green) and GR …
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Figure 4—source data 1
Raw data for generating Figure 4B–E.
- https://doi.org/10.7554/eLife.34135.012
Figure 5 with 1 supplement
Effect of acute prior stress on auditory-cued fear conditioning.
(A) The experimental design for prior restraint stress (RS) exposure and fear conditioning. Floxed GR and LAGRKO mice were exposed to a 20 min RS and fear-conditioned 1 hr later. Contextual and …
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Figure 5—source data 1
Raw data for generating Figure 5B–D.
- https://doi.org/10.7554/eLife.34135.015
Figure 5—figure supplement 1
Restraint stress- and auditory fear conditioning-induced release of corticosterone was comparable between floxed GR and LAGRKO mice.
(A) Plasma corticosterone levels 1 hr after RS exposure (n = 5). (B) Plasma corticosterone levels 90 min after RS exposure followed by AFC (n = 5). Data are presented as mean ± S. E. M.
Figure 6 with 1 supplement
Optogenetic manipulation of LTP or LTD at auditory inputs to the LA alters the effect of acute stress on auditory-cued fear memory.
(A) Experimental schedule for AAV injection, RS exposure, auditory-cued fear conditioning, and optical LTP or LTD induction. (B) Representative fluorescence image of areas expressing …
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Figure 6—source data 1
Raw data for generating Figure 6C–E.
- https://doi.org/10.7554/eLife.34135.018
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Figure 6—source data 2
Raw data for generating Figure 6F,G.
- https://doi.org/10.7554/eLife.34135.019
Figure 6—figure supplement 1
A putative model of LAGR-dependent mechanisms underlying alterations in synaptic plasticity during fear conditioning in basal condition or 1 hr after RS exposures.
In floxed GR mice, RS-induced release of corticosterone activates G protein-coupled postsynaptic membrane glucocorticoid receptors (mGRs), leading to the synthesis and release of endocannabinoid. …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers |
|---|---|---|---|
| Gene (Mus musculus) | Nr3c1 | Mouse Genome Informatics http://www.informatics.jax.org/marker/MGI:95824 | MGI:95824; NCBI Gene: 14815 |
| Gene (Mus musculus) | Grp | Mouse Genome Informatics http://www.informatics.jax.org/marker/MGI:95833 | MGI:95833; NCBI Gene: 225642 |
| Strain (C57BL/6-Tg(CAG-flpe) 36Ito/ItoRbrc) | CAG-FLPe mouse | RIKEN BioResource Center | RRID:IMSR_RBRC01834 |
| Strain (C57BL/6) | CAG-CAT-Z mouse | Proc Natl Acad Sci U S A 92: 160–164,1995 | |
| ES cell line | RENKA | Eur. J. Neurosci. 24: 2177–2190, 2006 | |
| Antibody | Rabbit anti-GR antibody | Santa Cruz Biotechnology Cat# sc-8992 | RRID:AB_2155784 |
| Antibody | Anti-NeuN antibody | Millipore Cat# MAB377 | RRID:AB_2298772 |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.34135.020
