Cerebral Cortex October 2010;20:2433--2441
doi:10.1093/cercor/bhp311
Advance Access publication January 15, 2010
Stress and Amygdala Suppression of Metaplasticity in the Medial Prefrontal Cortex
Gal Richter-Levin1,2,3 and Mouna Maroun1
1
Department of Neurobiology and Ethology, Faculty of Sciences, 2Department of Psychology and 3Institute for the Study of Affective
Neuroscience, University of Haifa, Haifa 31905, Israel
Address correspondence to Mouna Maroun, PhD. Email: mmaroun@psy.haifa.ac.il, mouna.maroun@gmail.com.
The term ‘‘metaplasticity’’ refers to the modulation of the ability to
induce synaptic plasticity of the form of long-term potentiation (LTP)
or long-term depression (LTD) following prior activation of the
synapses. While often electrophysiological manipulations are used to
demonstrate this phenomenon, prior behavioral manipulations such
as exposure to stress were also found to affect the ability to induce
LTP and LTD. Interestingly, amygdala stimulation was found to have
effects on subsequent LTP induction that resemble those of stress.
Here, we report that exposure to stress or basolateral amygdala
(BLA) stimulation induces a form of metaplasticity, which prevents
the ability of a second episode of stress or BLA activation to
suppress LTP in the ventral hippocampus-medial prefrontal cortex
(mPFC) pathway. This form of metaplasticity is N-methyl-D-aspartic
acid (NMDA)-dependent since the injection of the NMDA partial
agonist D-cycloserine prevented the inhibition of LTP induced by prior
exposure of stress or BLA activation. Furthermore, blocking NMDA
receptors by MK801 before the exposure to stress prevented the
ability of the emotional manipulation to inhibit the subsequent
modulation of plasticity, resulting in impaired LTP in the mPFC. Taken
together, these findings demonstrate a new form of NMDA-
dependent emotional metaplasticity in the ventral hippocampus-
mPFC pathway.
Keywords: D-cycloserine, hippocampus, LTP, MK801, priming
Introduction
‘‘Metaplasticity’’ was coined to describe modulation of the
ability to induce synaptic plasticity by a history of activity
within the involved network (Abraham and Bear 1996).
Employing different protocols of prior synaptic activation
affected the likelihood of inducing long-term potentiation
(LTP) or long-term depression (LTD) in the hippocampus
(Abraham et al. 2001; Zelcer et al. 2006) and cortex (Froc and
Racine 2004).
Heterosynaptic-induced metaplasticity was also described in
the hippocampus (Abraham and Goddard 1983; Doyère et al.
1997; Abraham et al. 2007). Specifically, prior amygdala
activation was found to affect the induction of LTP in the
hippocampus. Priming the basolateral amygdala (BLA) reduced
the threshold for LTP induction (Ikegaya et al. 1995), altered
the maximal level of potentiation (Akirav and Richter-Levin
1999; Vouimba and Richter-Levin 2005; Vouimba et al. 2007),
and promoted late-phase LTP (Korz and Frey 2003). These
effects are region specific since amygdala priming was found to
enhance LTP in the dentate gyrus (DG) while suppressing it in
CA1 (Vouimba and Richter-Levin 2005).
Exposure to stress was suggested to induce ‘‘behavioral’’
metaplasticity similar to that observed after amygdala priming
(Abraham and Tate 1997; Kim and Yoon 1998). Exposure to
stress suppressed the ability to induce LTP in the CA1 (Foy
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et al. 1987; Xu et al. 1997; Maroun and Richter-Levin 2003) and
enhanced LTP in the DG, similar to the differential effects of
amygdala priming on hippocampal subregions (Kavushansky
et al. 2006). Taken together, these observations raise the
possibility that the activation of the BLA mediates, at least in
part, the metaplastic effects of stress in the hippocampus.
Most studies addressing the metaplastic effect of stress on
LTP have mainly focused on the hippocampus. However,
plasticity such as LTP can easily be induced in the medial
prefrontal cortex (mPFC; Jay et al. 1995; Gurden et al. 2000;
Maroun and Richter-Levin 2003), and exposure to stress
suppresses LTP in the mPFC (Maroun and Richter-Levin 2003;
Rocher et al. 2004).
Several lines of evidence suggest that emotional arousal
activates the BLA and that this activation results in modulation
of memory-related processes in the hippocampus (for review,
see Cahill and McGaugh 1998; McGaugh 2000; Richter-Levin
and Akirav 2000; Roozendaal 2000; Abe 2001; Packard and
Cahill 2001).
The purposes of this study were to assess whether BLA
activation will mimic stress effects on LTP in the mPFC and to
examine the mechanisms by which BLA activation or exposure
to stress modulates LTP in the ventral hippocampus-mPFC
pathway.
We first established that both stress and BLA priming suppress
the ability to induce LTP in the mPFC. We further show that
exposure to behavioral stress or BLA stimulation induces a
form of metaplasticity, which prevents the ability of a second
episode of stress or BLA activation to suppress LTP in mPFC. We
finally show the dependency of this form of metaplasticity on
N-methyl-D-aspartic acid (NMDA) receptor activation.
Materials and Methods
Electrophysiology
Male Sprague Dawley rats (280--380 g) were anesthetized (with 40%
urethane and 5% chloral hydrate in saline, 0.5 ml/100 g, intraperito-
neally [i.p.]) and placed in a stereotaxic frame, with body temperature
maintained at 37 ± 0.5 °C. The procedures were performed in strict
accordance with University of Haifa regulations and National Institute
of Health (NIH) guidelines (NIH publication number 8023). In brief,
small holes were drilled into the skull to allow the insertion of
electrodes into the brain. A single recording microelectrode (glass, tip
diameter of 2--5 lm, filled with 2M NaCl, resistance of 1--4 M) was
slowly lowered into the mPFC (anteroposterior, 3.0--3.3 mm anterior to
bregma, 0.7--1.0 mm lateral, 3.8--4.8 mm below pial surface).
A bipolar 125-lm stimulating electrode was implanted in the area of
the CA1/subicular region of the ventral hippocampus (6.3--6.8 posterior
to bregma, 5.5 mm lateral, 4.0--5.8 mm below pial surface). An additional
stimulating electrode was lowered to the BLA (anteroposterior, –3 mm
relative to bregma, 5.0 mm lateral, –7.6 mm ventral). The evoked
responses were digitized (10 kHz) and analyzed using the Cambridge
Electronic Design (CED, Cambridge, UK) 1401+ and its Spike2 software.
Offline measurements were made of the amplitude of field postsynaptic
potentials (fPSPs) using averages of 5 successive responses to a given
stimulation intensity applied at 0.1 Hz. Test stimuli (monopolar pulses,
100 ls duration) were delivered at 0.1 Hz. After positioning the
electrodes, the rats were left for 30 min before commencing the
experiment.

LTP Induction
Theta burst stimulation (TBS) protocol: theta-like high-frequency
stimulation at 100 Hz to the ventral hippocampus (3 sets of 10 trains,
each train consisted of 10 pulses at 100 Hz, intertrain interval of 200

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ms, interset interval of 1 min).
For all the experiments, baseline responses were established by
means of delivering stimulation intensity (50--150 lA) sufficient to elicit
a response representing 25--30% of the maximal amplitude of the
evoked field potentials.
TBS was delivered at the same intensity and pulse duration as the test
stimuli during establishment of the baseline responses. Evoked field
potentials at the baseline intensity were recorded from the mPFC for
up to 60 min following the application of TBS. LTP was measured as an
increase in fPSP amplitude. Changes in the fPSP amplitude were
measured as a percentage change from the baseline.
TBS was applied in all groups within 1 hr 30 min following
anesthesia.

Amygdala Priming
The BLA was primed according to previous work (1 V, 50-ls pulse
duration, 10 trains of 5 pulses at 100 Hz, intertrain interval of 200 ms;
Vouimba and Richter-Levin 2005) either 30 s or 1 h before TBS was
applied to the ventral hippocampus.
Behavioral Stress Protocol
Stress was evoked by placing the rats on an elevated platform (EP) (12
3 12 cm) in a brightly lit room for 30 min (Xu et al. 1997; Maroun and
Richter-Levin 2003).
After the termination of the stressor, rats were immediately
anesthetized and taken for electrophysiological testing.
For the 2 episodes of exposure to stress, rats were placed on the EP
for 30 min and then taken to home cage for 30 min before being
reexposed to the EP stress for additional 30 min.
Drugs
D-cycloserine (DCS: a partial NMDA receptor agonist, 15mg/kg i.p) was
from Sigma (St Louis, MO). This dose was based on a previous work
showing facilitation of extinction of fear following systemic injection
(Ledgerwood et al. 2005).
The NMDA receptor antagonist, [1]-5-methyl-10,11-dihydro-5H-
dibenzo-[a,d]-cyclohepten-5,10-imine hydrogen maleate (MK801; Bio-
trend GmbH, Cologne, Germany), was dissolved in saline (0.1 mg/kg,
i.p.). This dose of MK801 is based on our previous work showing that it
does not impair the induction of LTP (Rosenblum et al. 1999).
The drugs were injected 30--40 min before the exposure to stress
(when applicable) or the application of TBS. Controls received
physiological saline at the same time as the drug groups.
Histology
Histological verification of the locations of the stimulating electrode in
the BLA was performed. After the electrophysiological testing, marking
lesions was made by passing anodal current (10 mA for 3 s). Only
animals that had their stimulating electrode in the BLA were included
(Fig. 1).
Statistical Analysis
Differences were determined using one-way or repeated measures
analysis of variance (ANOVA). All post hoc comparisons were made
using the least significant difference multiple comparison test.
Figure 1. A diagram depicting a coronal section of the rat brain (3.00 mm posterior
to bregma; Paxinos and Watson, 1998) showing electrode placements in the BLA.
Results
The Effects of Behavioral Stress and Amygdala Activation
on the Induction of LTP in the Ventral Hippocampus-
mPFC
All the groups received TBS to the ventral hippocampus, and
specifically, 4 groups were tested in this experiment: rats that
only received TBS (TBS only, n = 7), rats that were exposed to
EP and received TBS (EP, n = 5), rats with BLA priming 30 s
before TBS (BLA-30sec, n = 9), and rats with BLA priming 1 h
before the application of TBS (BLA-1hr, n = 6).
One-way ANOVA showed similar stimulus intensities applied
for all the groups (F3,23 = 1.01, not significant [n.s.]), and
a comparison between the groups using ANOVA with repeated
measures for the time points before the application of TBS did
not reveal a significant difference in fPSP amplitude (F3,23 =
0.77, n.s.) indicating a similar baseline. The average of the
amplitudes during baseline recording is presented in Table 1.
ANOVA with repeated measures on the levels of potentiation
at the different time points following the application of TBS
revealed that the 4 groups significantly differed (F3,23 = 7.12,
P = 0.001; Fig. 2) without a significant effect of post-TBS time or
significant interaction between treatment and time, indicating
that there was no difference between groups in this respect.
Post hoc analysis revealed that the TBS group was
significantly different from the other groups. The TBS group
had intact levels of LTP in the mPFC as a response to TBS
stimulation to the ventral hippocampus (133.6 ± 5.7%)
compared with the EP group (95.5 ± 8.4%, P < 0.002) to
the BLA-30sec (92.7 ± 6. 3%, P < 0.0001) and to the BLA-1hr
(105.1 ± 7.6%, P < 0.001).
Our data suggest that exposure to the EP stressor or
activation of the amygdala 30 s or 1 h before the application
of TBS to the ventral hippocampus inhibited the ability to
induce LTP in the mPFC. The data indicate that both EP stressor
and the specific protocol used in this study of electrical
activation of the BLA prior to TBS to the ventral hippocampus

2434 Emotional-Induced Metaplasticity in the Prefrontal Cortex d

Richter-Levin and Maroun
resulted in a form of metaplasticity that impaired LTP induction
in the mPFC (Fig. 2).
Stress and Amygdala Activation Effects on Metaplasticity
of Prior Experience in the Ventral Hippocampus-mPFC
Here, we examined how exposure to stress or electrical
activation of the BLA would affect the response of the mPFC to
a subsequent emotional experience.
Five groups were tested all of which received TBS: a group
that underwent 2 consecutive episodes of stress (each episode
of 30 min on the EP, separated by 30 min at home cage, EP-EP,
n = 5) and then received TBS, a group that underwent 2
consecutive amygdala priming episodes and then received TBS
(1 h and 30 s before TBS, BLA-BLA, n = 6), a group exposed to
the EP stress and BLA priming 30 s before TBS (EP + priming,
n = 6), a control group that received only TBS (TBS only, n = 6),
and an additional group that was exposed to a single episode of

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EP (EP, n = 4).
One-way ANOVA for the analysis of the stimulus intensities
Table 1 that were applied showed that there was no significant
The baseline amplitude of the fPSP in the mPFC in the different groups difference between the groups (F4,24 = 0.52, n.s.).
Group Baseline amplitude (mV) Comparison between the groups before the application of
TBS only 4.23 ± 0.63
TBS with repeated measures ANOVA did not reveal a significant
BLA-30sec 4.08 ± 1.06 difference in fPSP amplitude at any time point pre-TBS (F4,22 =
BLA-1hr 3.89 ± 0.37 1.24, n.s.) indicating a similar baseline. The average of the
EP 4.21 ± 0.97
EP-EP 3.89 ± 0.75 amplitudes during baseline recording is presented in Table 1.
BLA-BLA 3.88 ± 0.50 ANOVA with repeated measures following the application of
EP þ priming 3.62 ± 0.63 the TBS revealed significant differences between the groups
DCS 4.92 ± 0.83
DCS þ EP 4.33 ± 0.98 (F4,22 = 6.85, P = 0.001; Fig. 3). Post hoc analysis showed that
DCS þ BLA 4.21 ± 0.88 while the EP-EP, BLA-BLA, and the EP + priming groups
MK801 4.54 ± 0.72
MK801 þ EP þ priming 3.63 ± 0.65
exhibited normal and intact levels of LTP that were comparable
with the levels of the TBS-only group (TBS only: 129.7 ± 6.6%,
Note. Table 1 summarizes the amplitude of the fPSP of the different groups and shows that the EP-EP: 136.6 ± 7.2%, EP + priming: 129.1 ± 6.5%, BLA-BLA:
groups did not differ in their baseline amplitude, suggesting that the different manipulations did 138.14 ± 6.3%), the group that underwent a single episode of
not affect baseline transmission. The comparison between the relevant groups for each
experiment is detailed in the text.
stress had impairment of LTP (EP: 99.16 ± 6.1%, P < 0.001

Figure 2. Overall, 4 groups were tested: rats that only received TBS (TBS only, n 5 7), rats that were exposed to EP and received TBS (EP, n 5 5), rats with BLA priming 30 s
before TBS (BLA-30sec, n 5 9), and rats with BLA priming 1 h before the application of TBS (BLA-1hr, n 5 6). ANOVA analysis with repeated measures on post-TBS time points
revealed that the 4 groups significantly differed in the levels of potentiation following the application of TBS (F3,23 5 7.12, P 5 0.001). Exposure to behavioral stress or activation
of the BLA 30 s or 1 h before TBS to the ventral hippocampus inhibited LTP in the mPFC. Inset: representative average waveforms. Horizontal bar 5 10 ms, vertical bar 5 0.2 mV.
Arrow denotes the application of TBS.

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Figure 3. Five groups were tested: a group that underwent 2 consecutive episodes of stress (each episode of 30 min on the EP, separated by 30 min at home cage, EP-EP, n 5
5) and then received TBS, a group that underwent 2 consecutive amygdala priming episodes and then received TBS (1 h and 30 s before TBS, BLA-BLA, n 5 6), a group exposed
to the EP stress and the amygdala BLA priming 30 s before TBS (EP þ priming, n 5 6), a group that was exposed to a single episode of EP (EP, n 5 4), and a control group that
received only TBS (TBS only, n 5 6). ANOVA analysis with repeated measures on the post-TBS time points revealed significant differences between the groups (F4,22 5 6.85,
P 5 0.001). The group of a single exposure to EP had impairment in LTP, and the other groups with 2 episodes of BLA/EP or combined manipulations had intact levels of LTP.
Inset: representative average waveforms. Horizontal bar 5 10 ms, vertical bar 5 0.2 mV. Arrow denotes the application of TBS.

compared with all groups). There was no effect of time (post-
TBS) and the interaction between treatment and time was not
significant, indicating that there was no difference between
groups in this respect.
In the second experiment, we aimed to examine whether
blocking the NMDA receptors by injecting the NMDA
receptors antagonist MK801 would prevent the combined
effect of EP and BLA activation on the induction of LTP
(combined EP + BLA results in intact levels of LTP; Fig. 3).

The Role of the NMDA Receptor in Metaplasticity of
Emotional Modulation of LTP in the Ventral
Hippocampus-mPFC
The results of Experiments 1 and 2 show that while a single
episode of either activation of the BLA or exposure to the EP
stress inhibited LTP in the mPFC, 2 episodes of BLA activation
or 2 exposures to the EP, or the combination of both EP and
BLA activation resulted in intact levels of LTP. These results
suggest that electrical activation of the BLA or exposure to the
EP stress induced a form of metaplasticity that prevented the
ability of a subsequent experience—episode of stress or BLA
activation—to suppress LTP in the mPFC.
LTP in the mPFC was previously found to be NMDA
dependent (Jay et al. 1995; Maroun and Richter-Levin 2003),
and the involvement of the NMDA receptors in metaplasticity is
well established (Mockett et al. 2002; MacDonald et al. 2007).
Based on this, in the following experiment, we examined
whether this novel form of metaplasticity described above is
also NMDA dependent.
Two mirror experiments were conducted. In the first
experiment, the partial NMDA agonist DCS was used to assess
whether systemic activation of the NMDA receptors by the
injection of DCS could mimic the first episode either of the EP
stress or of BLA priming. To that end, rats were injected with
DCS and were either exposed to the EP or had the BLA activated.
The Effects of Activating the NMDA Receptors
Six groups were tested: rats that were exposed to the EP and
received TBS (EP, n = 7), rats that were BLA primed 30 s before
TBS to the ventral hippocampus (BLA-30sec, n = 6), rats with
the NMDA partial agonist (DCS, n = 5), rats that were injected
with DCS and placed on the EP and then TBS was applied
(DCS + EP, n = 5), rats that were injected with DCS and then
the BLA was primed 30 s before TBS to the ventral
hippocampus (DCS + BLA, n = 6), and a control group that
received only TBS (TBS only, n = 4).
The control groups received physiological saline injections
at the same time as the drug groups.
The behavior of saline- and DCS-treated rats on the EP (i.e.,
urination, defecation, freezing) was not different (data not
shown).
One-way ANOVA showed that similar stimulus intensities
were applied (F5,27 = 1.3, n.s.). Using ANOVA with repeated
measures on the fPSP amplitude before the application of TBS
did not reveal significant differences in fPSP amplitude (F5,27 =
0.26, n.s.) indicating a similar baseline. The average of the
amplitudes during baseline recording is presented in Table 1.
ANOVA with repeated measures on the time points
following the application of TBS showed significant differences
between the groups (F5,27 = 12.13, P < 0.0005; Fig. 4). The
interaction between treatment and time (post-TBS) was not

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Richter-Levin and Maroun
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Figure 4. Six groups were tested: rats that were exposed to the EP and received TBS (EP, n 5 7), rats that were BLA primed 30 s before TBS to the ventral hippocampus (BLA-
30sec, n 5 6), rats with the NMDA partial agonist (DCS, n 5 5), rats that were injected with DCS and placed on the EP and then TBS was applied (DCS þ EP, n 5 5), rats that
were injected with DCS and then the BLA was primed 30 s before TBS to the ventral hippocampus (DCS þ BLA, n 5 6), and a control group that received only TBS (TBS only,
n 5 4). The control groups received physiological saline injections at the same time as the drug groups. ANOVA with repeated measures analysis revealed significant differences
between the groups (F5,27 5 12.13, P \ 0.0005). The injection of DCS reversed the impairing effect of the EP or amygdala priming on LTP since the DCS þ EP and the DCS þ
BLA had intact levels of LTP comparable with those of TBS group. The DCS-alone group did not differ from the TBS group, indicating that DCS alone did not cause a further
enhancement in the potentiation levels beyond the control levels. The EP and the BLA-primed groups failed to induce LTP. Inset: representative average waveforms. Horizontal
bar 5 10 ms, vertical bar 5 0.2 mV. Arrow denotes the application of TBS.

significant, indicating that there was no significant difference
between groups in this respect.
The EP and the BLA-primed groups did not express LTP,
confirming the data from the previous experiment that both
stress and BLA priming impair the induction of mPFC LTP
(97.4 ± 6.5% and 90.3 ± 7%, respectively; P < 0.005 for both
compared with the TBS group). In contrast, the DCS reversed
the impairing effect of the EP or BLA priming since the DCS + EP
(122.5 ± 7.6%) and the DCS + BLA (144.5 ± 7%) did not differ in
the levels of potentiation from TBS-only group (137.79 ± 8.5%).
Similarly, the DCS group did not differ from the TBS-only
group (148.7 ± 7.7%), indicating that the DCS alone did not
cause a further enhancement in the potentiation levels beyond
the control levels.
The Effects of Blocking the NMDA Receptors
Four groups were tested: rats that received MK801 at a dose of
0.1 mg/kg (i.p.); we have previously shown that this dose does
not affect LTP in the DG of the hippocampus (Rosenblum et al.
1999; MK801, n = 5); rats that were exposed to the EP and then
had the amygdala primed 30 s before TBS to the ventral
hippocampus (EP + BLA, n = 6); rats that were injected with
MK801 and exposed to the EP and had their BLA primed 30 s
before TBS to the ventral hippocampus (MK801 + EP + BLA, n =
5), and control rats that had TBS only (TBS only, n = 5).
The control group received physiological saline injections at
the same time as the drug groups.
The behavior of saline- and MK801-treated rats on the EP (i.e.,
urination, defecation, freezing) was not different (data not
shown).
Similar stimulus intensities were applied (F3,17 = 0.8, n.s.), and
a comparison between the groups before the application of TBS
did not reveal a significant difference in fPSP amplitude (F3,17 =
1.70, n.s.) indicating a similar baseline. The average of the
amplitudes during baseline recording is presented in Table 1.
ANOVA with repeated measures on the time points
following TBS showed that the groups significantly differed
(F3,17 = 9.13, P < 0.005; Fig. 5), without significant differences
in time post-TBS (F < 1) or interaction between treatment and
time (F < 1).
Similar to previous data (Rosenblum et al. 1999), the
injection of the MK801 at this dose did not block the induction
of LTP, and its level was not statistically different from control
levels of LTP (122.9 ± 5.5% and 131.8 ± 5.4%, respectively, n.s.).
However, the injection of MK801 prior exposure to the EP
blocked stress-induced metaplasticity since following MK801,
the EP stress failed to block the impact of BLA priming on
mPFC LTP, and this group had impaired levels of LTP and was
significantly different from the 2 other groups (95.9 ± 5.5%
different from both groups, P < 0.005).
Discussion
The ventral hippocampus is reciprocally connected with the
BLA and mPFC (Groenewegen et al. 1990; Jay and Witter 1991),
suggesting an intensive interaction between these structures.
The BLA was found to modulate plasticity in the hippocampus
(Richter-Levin 2004). However, the ability of the BLA to
modulate plasticity in the ventral hippocampus-mPFC pathway
was not yet studied.

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Figure 5. Four groups were tested: rats that received MK801 at a dose of 0.1 mg/kg (i.p., MK801, n 5 5), rats that were exposed to the EP and then had the amygdala primed
30 s before TBS to the ventral hippocampus (EP þ BLA, n 5 6), rats that were injected with MK801 and exposed to the EP and had their BLA primed 30 s before TBS to the
ventral hippocampus (MK801 þ EP þ BLA, n 5 5), and control rats that had TBS only (TBS only, n 5 5). The control group received physiological saline injections at the same
time as the drug groups. ANOVA with repeated measures revealed that the groups significantly differed (F3,17 5 9.13, P \ 0.005). The injection of the MK801 at this dose did
not block the induction of LTP, and its level was not statistically different from control levels of LTP in the TBS group. However, the blockade of NMDA receptors prevented the EP
stressor from blocking the impact of BLA priming on mPFC LTP, resulting in impaired levels of LTP. Inset: representative average waveforms. Horizontal bar 5 10 ms, vertical
bar 5 0.2 mV. Arrow denotes the application of TBS.

The present report demonstrate that similar to behavioral
stress, electrical BLA activation suppressed the ability to
induce LTP in the mPFC (Maroun and Richter-Levin 2003;
Rocher et al. 2004). Priming the BLA (30 s prior to TBS to the
ventral hippocampus) or spaced activation (1 h prior to TBS
to the ventral hippocampus, a time interval similar to that of
the exposure to stress) suppressed LTP in the mPFC,
indicating that suppression of LTP is not due to the interval
between the activation of the BLA or exposure to stress and
TBS but rather the fact that the amygdala was activated. These
results further strengthen the view that the effects of stress
on plasticity are mediated, at least in part, by amygdala
activation (Richter-Levin 2004). Furthermore, they show that
this pathway expresses opposite effects to those observed in
the DG (Ikegaya et al. 1995; Akirav and Richter-Levin 1999)
and similar to those observed in the CA1 subregion (Maroun
and Richter-Levin 2003; Vouimba and Richter-Levin 2005;
Vouimba et al. 2007).
The impairment of mPFC-LTP following stress is congruent
with data showing that exposure to stress has detrimental
effect on mPFC-dependent behaviors, such as working memory
tasks (e.g., Murphy et al. 1996; Mizoguchi et al. 2000).
Interestingly, adrenalectomy (Mizoguchi et al. 2004),
acute or chronic corticosterone treatment (Bardgett et al.
1994; Roozendaal et al. 2004), or infusion of a glucocorticoid
receptor agonist directly into certain subregions in the
mPFC (Roozendaal et al. 2004) also impaired performance on
working memory tasks. These effects are reminiscent of the
findings that adrenalectomy or corticosterone administra-
tion or exposure to stress produced dendritic retraction in
mPFC (Wellman 2001; Izquierdo et al. 2006; Cerqueira et al.
2007).
Furthermore, other mPFC-dependent memory tasks, like
extinction of fear conditioning were also found to be impaired
following exposure to behavioral stress (Izquierdo et al. 2006;
Akirav and Maroun 2007; Akirav et al. 2009). Interestingly, the
suppression of LTP in the PFC by stress was suggested to underlie
stress-induced impairment of extinction (Herry and Garcia 2002;
Maroun and Richter-Levin 2003; Maroun 2006). Stress-induced
impairment is suggested to be of relevance to anxiety disorders,
such as post-traumatic stress disorder (Izquierdo et al. 2006;
Akirav and Maroun 2007; Akirav et al. 2009).
The main question of the current study was whether prior
activation would include an additional type of metaplasticity or
in other words whether electrical stimulation of the BLA or
behavioral exposure to the stressor could induce changes that
would affect at later stages the ability to induce further changes
in the mPFC.
Indeed, the results show that prior exposure to the EP stress
suppressed the ability of a subsequent stress exposure to
inhibit LTP induction. Similarly, prior BLA activation also
suppressed the ability of subsequent BLA priming to inhibit
LTP induction in the mPFC. Interestingly, prior stress exposure
also prevented the ability of amygdala priming to inhibit LTP
induction in the mPFC, suggesting that similar mechanisms
were activated by prior stress exposure and prior electrical
activation of the BLA (the effects are summarized in Table 2).
The results clearly demonstrate the formation of a type of an
emotional metaplasticity that inhibits the response of the mPFC
to a subsequent emotional event for at least 40 min.

2438 Emotional-Induced Metaplasticity in the Prefrontal Cortex d

Richter-Levin and Maroun
Table 2
The effect of the different manipulations on the ability to induce LTP in the ventral hippocampus-
mPFC pathway
Group LTP
TBS only Normal LTP
BLA-30sec
BLA-1hr
EP
EP-EP Normal LTP
BLA-BLA Normal LTP
EP þ priming Normal LTP
DCS Normal LTP
DCS þ EP Normal LTP
DCS þ BLA Normal LTP
MK801 Normal LTP
MK801 þ EP þ priming
The manipulations of prior activation either behaviorally by
exposure to stress or the electrical activation of the BLA did
not affect baseline transmission as no effect was observed on
the fPSP amplitude. Furthermore, Mockett and Hulme (2008)
suggest that the induction of a metaplastic state through
synaptic activation, while most easily observed when no
change in basal synaptic transmission, is induced and the
critical element is the modification of the response to later
synaptic stimulation compared with that of nonprimed path-
ways.
Caution is, however, appropriate. Although there were no
obvious alterations in baseline transmission, with our present
sample sizes (ranging from 4--9 per group for each compari-
son), we cannot exclude the possibility that differences may
have occurred following the different manipulations.
The finding that prior exposure to stress would ‘‘protect’’
from subsequent aversive effects of stress is at first glance
counterintuitive. However, the emotional manipulations ap-
plied here cannot be considered traumatic. The current
findings should be looked at in the context of responses to
emotional challenges within the range of a normative experi-
ence. We have previously demonstrated that exposure of rats
to a mild stress resulted in reorganization of maps of activation
of relevant brain areas, including the amygdala and hippocam-
pus (Akirav et al. 2001; Kogan and Richter-Levin 2008). Such
remapping of activation was not associated with impaired
performance in a learning task but rather with alterations of the
characteristics of the formed memory that could be described
as shifts in learning strategies (Kogan and Richter-Levin 2008).
The exposure to a single mild stress experience may be seen as
potentially threatening and thus leading to the suppression of
LTP in the mPFC and a parallel increased activation of the
amygdala. Repeated exposure to such a mild stressor without
significant aversive outcomes should mark this event as less
relevant and thus may lead to a shift to a less emotional map of
activation that includes the full involvement of mPFC plasticity
in the learning.
In line with this view, it is expected that a traumatic
experience will suppress LTP in the mPFC even when it
precedes an additional emotional experience. The current
findings should thus be seen within the context of emotional
learning. The form of metaplasticity described here contrib-
utes to our understanding of the regulation of adaptive
behavior like extinction of fear conditioning (Morgan et al.
1993; Milad and Quirk 2002; Akirav et al. 2006; Akirav and
Maroun 2007).
The Role of NMDA Receptors in Metaplasticity
The involvement of the NMDA receptor in some forms of LTP
and metaplasticity is well established (Mockett et al. 2002;
No LTP MacDonald et al. 2007).
Here, we examined whether NMDA activation is also
Inhibition of LTP required for this emotional type of metaplasticity. This was
Inhibition of LTP examined in 2 mirror sets of experiments; activation of
Inhibition of LTP
NMDA receptors by DCS induced metaplastic effects similar
Downloaded from https://academic.oup.com/cercor/article/20/10/2433/317108 by UNIVERSITAT DE LLEIDA user on 05 September 2023
to those of prior exposure to stress or to BLA priming, that is,
DCS blocked the ability of exposure to behavioral stress or
BLA manipulations to inhibit LTP induction in the mPFC.
Complementary to that, blocking of the NMDA receptors by
MK801 before the exposure to stress prevented the ability of
Inhibition of LTP
this emotional manipulation to inhibit subsequent emotional
modulation of plasticity and hence resulting in impaired LTP
in the mPFC. When the same dose of MK801 was
administered alone, it did not affect LTP in the mPFC,
consistent with our previous work that this dose does not
affect LTP in the DG of the hippocampus (Rosenblum et al.
1999).
These results show that MK801 prevented the ability of the
emotional manipulation to inhibit subsequent modulation of
plasticity, resulting in impaired LTP in the mPFC. (The main
effects of the different manipulations are summarized in Table
2.) The results may suggest that the threshold of affecting
metaplasticity might be lower than synaptic plasticity as was
previously suggested that metaplasticity could also occur when
priming stimulations fail to induce persistent changes in
synaptic plasticity in the hippocampus (Mockett and Hulme
2008; Xu et al. 2009).
The facilitatory effect of DCS has been previously reported.
Specifically, it was shown that a single dose of DCS injected 24
h post head injury prevented impairment in LTP in the
hippocampus and rescued the cognitive impairments associ-
ated with the trauma (Yaka et al. 2007). Consistent with that,
NMDA receptor activation through its glycine-binding site
rescued age-related deficits in LTP (Billard and Rouaud 2007).
Furthermore, it was previously shown that DCS administra-
tion enhanced the extinction of conditioned fear (Walker et al.
2002; Ledgerwood et al. 2003, 2005; Akirav et al. 2009) and
facilitated fear extinction in patients with anxiety disorders
when it is combined with exposure therapy but not when it is
given alone (Ressler et al. 2004; Hofmann et al. 2006). In
contrast, administration of DCS with no exposure therapy is
not effective (Heresco-Levy et al. 2002). This latter finding is
consistent with our finding that DCS alone did not cause
further enhancement of the magnitude of LTP. We have
recently found that when DCS before the exposure to the EP
stressor prevents the impairing effect of stress on extinction
(Akirav et al. 2009).
Taken together, these findings report a novel form of
emotional metaplasticity in the hippocampus-PFC and show
that the activation of this type of emotional metaplasticity is
NMDA dependent.
These mechanisms of metaplasticity might be potential
targets to treat cognitive impairments associated with expo-
sure to stress.
Funding
National Institute for Psychobiology to M.M.
Cerebral Cortex October 2010, V 20 N 10 2439
Notes
Conflict of Interest : None declared.
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