Reconsolidation of declarative memory in humans

Cecilia Forcato,
1
Valeria L. Burgos,
2
Pablo F. Argibay,
2
Victor A. Molina,
3
Mara E. Pedreira,
1,4
and Hector Maldonado
1,4,5
1
Laboratorio de Neurobiologa de la Memoria, Departamento de Fisiologa y Biologa Molecular y Celular, IFIBYNE-CONICET,
Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina;
2
Unidad de Ciencias
Cognitivas, Instituto de Ciencias Bsicas y Medicina Experimental, Hospital Italiano de Buenos Aires, C1181ACH Buenos Aires,
Argentina;
3
Departamento de Farmacologa Facultad de Ciencias Qumicas, Universidad Nacional de Crdoba,
5000 Cordoba, Argentina
The reconsolidation hypothesis states that a consolidated memory could again become unstable and susceptible to
facilitation or impairment for a discrete period of time after a reminder presentation. The phenomenon has been
demonstrated in very diverse species and types of memory, including the human procedural memory of a motor
skill task but not the human declarative one. Here we provide evidence for both consolidation and reconsolidation in
a paired-associate learning (i.e., learning an association between a cue syllable and the respective response syllable).
Subjects were given two training sessions with a 24-h interval on distinct verbal material, and afterward, they
received at testing two successive retrievals corresponding to the first and second learning, respectively. Two main
results are noted. First, the first acquired memory was impaired when a reminder was presented 5 min before the
second training (reconsolidation), and also when the second training was given 5 min instead of 24 h after the first
one (consolidation). Second, the first retrieval proved to influence negatively on the later one (the retrieval-induced
forgetting [RIF] effect), and we used the absence of this RIF effect as a very indicator of the target memory
impairment. We consider the demonstration of reconsolidation in human declarative memory as backing the
universality of this phenomenon and having potential clinical relevance. On the other hand, we discuss the
possibility of using the human declarative memory as a model to address several key topics of the reconsolidation
hypothesis.
The reconsolidation hypothesis states that a consolidated
memory could again become susceptible to facilitation or impair-
ment for a discrete period of time after retrieval (Nader et al.
2000a; Sara 2000a). It was initially supported by results obtained
with rodents and then confirmed with animals belonging to very
different species, such as the crab, chick, fish, and freshwater
snail (Anokhin et al. 2002; Eisenberg et al. 2003; Pedreira and
Maldonado 2003; Sangha et al. 2003). The range was extended to
humans, including the procedural memory of a motor skill task
(Walker et al. 2003); however, the reconsolidation hypothesis
has not yet been demonstrated for the declarative memory. This
demonstration would substantially back the universality of the
phenomenon, but what is more, would open the possibility of
discussing the reconsolidation hypothesis from the viewpoint of
the declarative memory, that is, from a type of memory that is
the hallmark of man (Dudai 2002). Contentious or still conjec-
tural topics of the reconsolidation hypothesis such as its func-
tionality (Dudai and Eisenberg 2004; Debiec et al. 2006; Sara and
Hars 2006), or the reminders requirements (Pedreira et al. 2004),
would surely gain a new perspective of analysis.
In most of the precedent studies with diverse animal species,
memory reconsolidation is demonstrated by the amnesic effects
induced by the administration of blockers, such as the protein
synthesis inhibitors or -blockers (Przybyslawski et al. 1999;
Nader et al. 2000a), or also by the learning of a new memory
(Walker et al. 2003; Boccia et al. 2005), after the presentation of
a reminder. The current method to demonstrate such amnesic
effects is by disclosing at testing a defective retrieval of the target
memory. The common sense of William James had already
pointed to the fact that the way to study memory is through its
retrieval, saying that the only proof of its retention is that it can
be recalled (James 1890; Sara and Hars 2006). However, this di-
rect method of evaluation may be sometimes misleading or in-
applicable, mainly when the declarative memory is involved. In
fact, memories are not stored in isolation from other memories
but integrated into complex associative networks (Levy and
Anderson 2002; Berman et al. 2003; Debiec et al. 2006), and then
the activation of related traces may interfere with the expression
of the desired retrieval, as has been documented over a century of
research (McGeoch 1932; Postman 1971; Anderson and Neely
1996). In other words, a faulty retrieval at testing may be due to
either problems in encoding storage or simultaneous retrieval of
related information (Mayes and Downes 1997), and then this
lack of specificity robs this direct method of its value to disclose
deficits in memory.
In the present study, we are proposing an alternative
method, based on the forgetting effect that the retrieval of the
target memory could have on the recall of related memories. This
effect, termed retrieval-induced forgetting (RIF) (Anderson et al.
1994; MacLeod and Macrae 2001), shows that the act of remem-
bering can temporarily block a late retrieval of other memory, or
more specifically, the expression of its retrieval. For this effect to
be possible, the inducing memory must be intact, and therefore,
the absence of RIF might become a good indicator of a defective
target memory. That is, we are proposing that memory deficits
were not revealed by the flaw of its own retrieval but by the good
retrieval of a related memory.
Here we demonstrate consolidation and reconsolidation in
paired-associate learning (i.e., an association between a cue syl-
lable and the respective response syllable). Subjects were given
4
These authors contributed equally to this work.
5
Corresponding author.
E-mail [email protected]; fax 54-11-45763384.
Article is online at http://www.learnmem.org/cgi/doi/10.1101/lm.486107.
Research
14:295303 2007 by Cold Spring Harbor Laboratory Press ISSN 1072-0502/07; www.learnmem.org Learning & Memory 295
www.learnmem.org
two training sessions, with an interval of 24 h, during which they
learned two distinct verbal materials. Afterward, they received at
testing two successive retrievals: the first one corresponding to
the first learning, and the second retrieval to the second one. The
first memory was impaired when the second learning occurred
immediately after the first one (consolidation) or up to 6 h after
the reminder presentation (reconsolidation). In both cases, the
memory impairment was disclosed not because the first retrieval
was faulty but because there was no RIF (Levy and Anderson
2002).
Results
The paired-associate memory
This first section of results deals with paired-associate learning
experiments (i.e., an association between a cue syllable and the
respective response syllable).
An analysis of the interaction between retrievals at testing: The
L2-training given 24 h after L1-training fails to impair consolidation
In the first experiment subjects were trained on day 1 with L1
and on day 2 with L2, to be finally tested on Day 3, either with
L1-testing (TL1) followed by L2-testing (TL2) or the other way
around (Fig. 1). The purpose of this experiment was focused on
the study of possible interferences between L1 and L2 retrievals at
testing session and on evaluating their use as a method to dis-
close deficits in the L1 memory, i.e., the memory acquired during
the first training with L1 (for term definitions, see Table 1).
The expression of the L1-retrieval at the first phase of testing is reduced
by the interference of simultaneous recruitment of items from the L2 memory
and/or by a loss of the L1-memory impairment
Results shown in Figure 1 (right panels) correspond to six retriev-
als at the testing session of the respective groups A, B, L1-CTL,
and L2-CTL, analysis of variance (ANOVA); F
(5,54)
= 10.546,
P < 0.001. If the analysis is confined to phase 1 of groups A and
B, we find that the L1-retrieval at phase 1 (group A) had a number
of errors significantly higher than that of the respective control
L1-CTL (P < 0.001), whereas the performance for the L2-retrieval
at phase 1 (group B) proved to be as good as that for the L2-CTL
(P = 0.546). The poor performance during the L1-retrieval at
phase 1 could be explained in terms of an interference from
retrieval of L2 on the expression of the L1-retrieval (L2 L1),
namely, the retrieval of the L1-memory simultaneously recruits
items from the L2-memory and these items interfere with the
expression of the L1-retrieval (retrieval interference) (McGeoch
1932). On the other hand, the good performance during the
L2-retrieval at phase 1 indicates absence of interference in the
opposite direction (L1 L2). This disparity is attributed to a dif-
ference between the memory strength for L1 and that for L2, due
to the fact that the former was acquired 48 h and the latter 24 h
before the test session (Ebbinghaus 1885; Wixted and Ebbesen
1997). Therefore, the strongest memory interferes with the re-
trieval of the weaker one but not vice versa.
However, an alternative explanation of the poor perfor-
mance of L1-retrieval at testing could be offered. The poor ex-
pression of the L1-retrieval at first phase of testing may be due
not only to the simultaneous recruitment of the L2 memory but
also (and/or) by a loss of the L1-memory, that is, by an impair-
ment of the L1-memory consolidation because of the second
training with L2.
The expression of the L2-retrieval at phase 2 of testing is negatively influenced
by the previous L1-retrieval at phase 1
If the analysis of results from Experiment 1 (Fig. 1) is now con-
fined to those of the L2-retrieval, we find a faultless expression of
L2-retrieval at phase 1 (group B) but a poor one at phase 2 (group
A), in comparison with L2-CTL (P = 0.516 and P < 0.001, respec-
tively). This difference indicates that regardless of whether the
retrieval of a memory is preceded by that of a related memory,
the first retrieval influences negatively the expression of the sec-
ond one, which represents an instance of RIF (Anderson et al.
1994; Levy and Anderson 2002). Since this effect depends on the
integrity of the memory first recalled (Anderson et al. 1994), the
result indicates that the L1 memory is intact, although its expres-
sion was reduced by the interference of the simultaneous re-
trieval of L2 memory. Therefore, we conclude that the second
training given 24 h after the first one does not impair the con-
solidation of the L1-memory.
The absence of the RIF effect is a specific indicator of deficits in the target
memory
The preceding analysis of results was based on the postulation
that the L1-retrieval at the first phase of testing not only inter-
feres with its own expression due to the simultaneous recruit-
ment of L2 items, but it also interferes, provided that the L1-
memory was intact, with the expression of the later L2-retrieval
(RIF effect). In the cases of L1-memory impairment, as is expected
to be found in the following series of experiments aimed at
studying impairments of L1-memory consolidation and recon-
solidation, a double effect should be expected: (1) a defective
expression of the L1-retrieval at phase 1 and (2) a reduction or
elimination of the RIF. The first effect cannot be distinguished
from that of the interference due to the simultaneous recruit-
ment of L2 items, whereas the second one specifically reveals
memory impairment. That is, a poor per-
formance for L1-retrieval at testing may
be interpreted as loss of L1-memory and/
or interference with its own expression
by simultaneous recall of L2 items; in-
stead, the absence of RIF is only regarded
as an expression of deficits in the L1-
memory. Namely, the absence of RIF,
but not the failure of L1-retrieval, is spe-
cific of L1-memory impairment.
Keeping in mind the above consid-
erations, results of the next series of ex-
periments will be evaluated by the RIF
effect from the L1-retrieval at phase 1 on
the expression of L2-retrieval at phase 2.
A number of errors for L2-retrieval at
phase 2 (panel A) higher than that of the
L2-CTL would indicate no impairment
Figure 1. Analysis of interaction between retrievals at testing. (Left) Groups (n = 10): L1-CTL, L1-
control; L2-CTL, L2-control; A, Group A, B, Group B; L1, L1-training; L2, L2-training; TL1, L1-testing;
TL2, L2-testing; 5m, the 5-min interval between testings. Phase 1, the first list tested (TL1 or TL2);
Phase 2, the second list tested (TL1 or TL2). L1-CTL and L2-CTL were evaluated on Day 3 with TL1 or
TL2, respectively. Group A was evaluated with TL1 on phase 1 and TL2 on phase 2; Group B was
evaluated with TL2 on phase 1 and TL1 on phase 2. (Right) Mean of total errors SEM on Day 3. Black
bars, TL1 performance; white bars, TL2 performance. ***, P < 0.001.
Forcato et al .
296 Learning & Memory
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of the L1-memory. In contrast, a performance for L2-retrieval at
phase 2 similar to that of the L2-CTL would indicate a deficit of
the L1-memory.
The L2-training given 5 min after L1-training impairs consolidation
of L1-memory
The purpose of the next experiment was to explore whether the
consolidation of the target memory (the L1-memory) could be
affected by giving the L2-training on Day 1 immediately after the
L1-training. Therefore, the protocol of this experiment (Fig. 2)
was similar to above except for the interval between trainings,
which in this case was only 5 min. Results of the testing session
on Day 3 are exhibited in Figure 2 (right panels) (ANOVA,
F
(5,54)
= 11.5, P < 0.001). The analysis of data corresponding to
group A discloses a significant difference between mean errors of
L1-retrieval at phase 1 and that of the L1-CTL (P < 0.002) but not
between L2-retrieval at phase 2 and the L2-CTL (P = 0.323). This
picture of results is indicative of an absent or insufficient RIF
effect as a consequence of deficits in the L1-memory. On the
other hand, the mean errors of L2-retrieval at phase 1 (group B)
was similar to that of L2-CTL (P = 0.710), a result that was ex-
pected since the impairment of L1-memory rules out the possi-
bility of an interference from the L1-retrieval (ANOVA,
F
(5,54)
= 5.091, P < 0.001).
In summary, the L2-training given 5 min after the L1-
training, impairs the consolidation of the memory previously
acquired. The deficit of L1-memory consolidation is specifically
disclosed by the absence or reduction of the RIF effect.
Study of the effect of L2-training given at diverse intervals
after reminder on reconsolidation of L1-memory
The following experiments were aimed at testing the reconsoli-
dation phenomenon (Fig. 3). In Experiment 1, subjects were
trained with L1 on Day 1 and with L2 on Day 2, 5 min after the
reminder presentation. The reminder trial included the specific
context of L1-training plus the presentation of one cue syllable,
which was abruptly interrupted after 2 sec of exposure, thus not
allowing the subject to write down the response syllable. Results
of test session on Day 3 (ANOVA, F
(5,54)
= 10.546, P < 0.001) were
closely similar to those obtained above as to memory consolida-
tion. The analysis of data of group A reveals a significant differ-
ence between mean errors of L1-retrieval at phase 1 and that of
the L1-CTL (P < 0.001) but not between L2-retrieval at phase 2
and the L2-CTL (P = 0.255). This picture of results is indicative of
an absent or insufficient RIF effect as a consequence of deficits in
L1-memory. On the other hand, results in group B repeat those
obtained throughout this paper, namely, the mean errors of L2-
retrieval at phase 2 were as that of the L2-CTL (P = 0.568) and the
performance of L1-retrieval at phase 2 was higher than that of the
L1-CTL (P < 0.001).
The remaining two experiments of this series were carried
out with the goal of pointing out the time window of the L1-
memory reconsolidation. The results obtained with L2-training
given 6 h after reminder (Fig. 3, Experiment 2) (ANOVA,
F
(5,54)
= 13.038, P < 0.001) showed a significance difference
(P < 0.002) for L1-retrieval at phase 1 vs. L1-CTL, and no differ-
ence (P = 0.214) for L2-retrieval at phase 2 vs. L2-CTL. These find-
ings were similar to those obtained with L2-training tested 5 min
after reminder (Fig. 3, Experiment 1), disclosing no RIF effect and
then a weakened L1-memory. Instead, results with the L2-
training given 10 h after reminder (Fig. 3, Experiment 3), showed
a significant difference for L1-retrieval at phase 1 vs. L1-CTL
(P < 0.001), and also for L2-retrieval at phase 2 vs. L2-CTL
(P < 0.004), indicating an RIF effect and L1-memory intact.
Therefore, the L2-training given 5 min or 6 h after present-
Table 1. Definitions of terms
Term Definition
Retrieval-induced
forgetting (RIF)
Refers to the phenomenon in which the
act of remembering some material
disrupts temporally the retrieval
expression of related material (Anderson
et al. 1994).
Retrieval interference Refers to the phenomenon in which the
act of remembering some material
recruits simultaneously items from
related material producing an
impairment in the expression of the
desired retrieval (McGeoch 1932).
Paired-associate memory A memory generated by a verbal learning
consisting of the association between a
cue syllable and a response syllable.
Prediction memory A memory generated by the association
between a specific context and the
verbal learning.
Context period Initial period of time of each trial where
the context is formed. It consists of a
fixed sequence of light, image, and
sound.
Specific context Combination of light, image, and sound
that is always followed by the verbal
learning.
Syllable period Period of time where the nonsense
syllables are presented.
Cue-response syllables Pairs of nonsense syllables of the verbal
learning.
Actual trial It includes the specific context followed by
the syllable presentation.
Fake trial It includes a context that is never followed
by the syllable presentation.
L1-training (or L1) It consists of 10 actual trials interspersed
with 22 fake trials separated by a 3-sec
intertrial interval. Each actual trial
includes the List 1 presented with the
respective specific context.
L2-training (or L2) It consists of 10 actual trials interspersed
with 22 fake trials separated by a 3-sec
intertrial interval. Each actual trial
includes the List 2 presented with the
respective specific context.
L1-memory Memory acquired by the L1-training (the
target memory).
L2-memory Memory acquired by the L2-training.
Reminder Trial that includes the specific context of
L1-training plus the presentation of one
cue syllable, which is abruptly
interrupted.
TL1 (or L1-Testing) Test of the L1 memory consisting of four
actual trials interspersed with eight of
the respective fake trials.
TL2 (or L2-Testing) Test of the L2 memory consisting of four
actual trials interspersed with eight of
the respective fake trials.
Phase 1 Period of the test session where one of the
two lists (L1 or L2) is first retrieved.
Phase 2 Period of the test session where one of the
two lists (L1 or L2) is retrieved 5 min
after phase 1.
L1-retrieval Retrieval of the prediction and
paired-associated memory
corresponding to the L1 memory.
L2-retrieval Retrieval of the prediction and
paired-associated memory
corresponding to the L2 memory.
Group A (A) Experimental groups that are tested for L1
in phase 1 and for L2 in phase 2.
Group B (B) Experimental groups that are tested for L2
in phase 1 and for L1 in phase 2.
L1-CTL (or L1-control) Control group that is only trained and
tested with L1 and TL1, respectively.
L2-CTL (or L2 control) Control group that is only trained and
tested with L2 and TL2, respectively.
Memory reconsol i dati on i n humans
Learning & Memory 297
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ing the reminder impairs the memory of L1 acquired 24 h before.
On the contrary, the same training given 10 h after the reminder
leaves the first memory intact, suggesting a period of reconsoli-
dation longer than 6 h and shorter than 10. This period appears
longer than that generally estimated for consolidation but the
comparison is not possible here since we have not investigated
the temporal window for consolidation. The deficit of L1-
memory in the two former experiments is specifically disclosed
by the absence or reduction of the RIF effect.
The impairing effect of the L2-training
on the target memory reconsolidation lasts
at least for 48 h
In order to test the disruptive effect on
memory reconsolidation, 48 h after the
L2-training, the following two experi-
ments were performed (Fig. 4). The pro-
tocol of Experiment 1 with groups A, L1-
CTL, a nd L2- CTL, ANOVA,
F
(3,36)
= 10.355, P < 0.001, was similar to
the one shown above in Figure 1; how-
ever, the test session was carried out 72 h
rather than 48 h after L1-training. Results
of group A (Fig. 4, Experiment 1) again show a RIF effect since the
number of errors for L1-retrieval at phase 1 and L2-retrieval at phase
2 were higher than those of their respective control groups, L1-CTL
(P < 0.001) and L2-CTL (P < 0.002). Thus, the RIF is disclosed at a
test session carried out 72 h after the first training which denotes an
enduring L1-memory.
The protocol of Experiment 2 (Fig. 4) includes group A, L1-
CTL, and L2-CTL and a reminder placed 5 min before the L2-
training. The pattern of results, (ANOVA, F
(3,36)
= 3.869,
P < 0.017) proved to be closely similar to
that of Figure 3, Experiment 1, since the
mean error of L2-retrieval at phase 2 was
similar to that of the L2CTL (P = 0.153),
whereas that of L1-retrieval at phase 1
resulted higher than the mean error of
the respective L1-CTL (P < 0.001). There-
fore, the absence of RIF is shown not
only at the test session given at 48 h, but
also 72 h after the first training. This last-
ing absence of RIF effect discloses an en-
during deficiency in the L1-memory,
namely, the demonstration that the dis-
rupting effect on the memory reconsoli-
dation persists no only for 24 h, but for 48
h after post-reminder L2-training.
The impact of the context period
on the paired-associate learning
and the uniformity of trainings
The control groups, L1-CTL and L2-CTL,
evidenced very good retention of the
paired-associate memory up to at least 4
d after the verbal material was acquired.
The range of mean errors per retrieval
was between 0.8 and 1.0 out of 20. We
attempt here to determine to what ex-
tent the inclusion of a period of context
formation before syllables presentation
could have improved the performance
of the subjects. A first group of 10 sub-
jects (the context group) was given 10
L1-actual trials, each with 20 sec of con-
text period (red light + New York
City + jazz) followed by the syllables pe-
riod, and separated by a 3-sec intertrial
interval. A second group of 10 subjects
(the no-context group) started the syl-
lables period 20 sec after the beginning of
the trial but with no context formation;
therefore, they were illuminated only
from the keyboard lamp during the entire
trial. Although both groups were given
Figure 2. Effect of an interfering training on memory consolidation. (Left) Groups (n = 10): controls,
A, B, L1, L2, TL1, TL2, Phase 1, and Phase 2, as in Figure 1; 5m, the 5 min between trainings or between
testings. (Right) Mean of total errors SEM on Day 3. **, P < 0.01.
Figure 3. Effect of an interfering training on memory reconsolidation. (Experiment 1) (Left) Groups
(n = 10): controls, A, B, L1, L2, TL1, TL2, Phase 1, and Phase 2, as in Figure 1; R, the reminder trial; 5m,
the 5 min between the reminder trial and L2, or between testings. (Right) Mean of total errors SEM
on Day 3. ***, P < 0.001. (Experiment 2) (Left) Groups (n = 10): controls, A, B, L1, L2, TL1, TL2, Phase
1, and Phase 2, as in Figure 1; 6h, the 6 h between the reminder trial and the L2 training; 5m, the 5
min between testings; R, as in Experiment 1. (Right) Mean of total errors SEM on Day 3. **, P < 0.01.
(Experiment 3) (Left) Groups (n = 10): controls, A, B, L1, L2, TL1, TL2, Phase 1, and Phase 2, as in Figure
1; 10h, the 10 h between the reminder trial and the L2 training; 5m and R, as in Experiment 1. (Right)
Mean of total errors SEM on Day 3. **, P < 0.01; ***, P < 0.001.
Forcato et al .
298 Learning & Memory
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the same paired-associate training (i.e., the same pairs of cue syl-
lable and response syllable), there was a significant difference in the
number of response errors at L1-testing between both groups
(ANOVA, F
(1,18)
= 6.015, P = 0.027) (Fig. 5A).
Moreover, we try to explore the degree of uniformity of the
performances at paired-associate trainings throughout the experi-
ments of this study. For this purpose, we compare the mean errors
for the last four actual trials of the training tail (Fig. 5B) correspond-
ing to the control groups of the seven experiments (L1-CTL and
L2-CTL). An ANOVA of these values across experiments and type of
training (L1, L2), indicated that there was no experiment or type of
training effect or interaction (P = 0.961, 0766, and 0.998, respec-
tively). That is, there was a noticeable uniformity in the acquisition
course of the paired-associate memory through experiments and
type of training.
The prediction memory
Over training, subjects learn to identify the specific context,
namely, the only light-image-sound sequence that is followed by
the syllables display. The level of the prediction memory was
assessed by the number of proper responses in pressing the ex-
pectancy keys (YES or NO) in both actual and fake trials. We
compare the learning curves of prediction with those of learning
to respond to a cue syllable. In Figure 5B, we show this contrast
for the L1-training of the experiment shown above in Figure 1.
The successive regression analysis (Lozada et al. 1990) revealed
different values for both asymptote and slope for either the pre-
diction or the paired-associate learning (asymptotic trial: 8 and
21, respectively; slope value: 12.867 and 4.898, respectively).
This result is consequently shown for all the 38 training curves of
the paper.
The mean of accumulative errors corresponding to either
the training tail or the retrieval (four actual trials + eight fake
trials) is practically equal to zero for the prediction memory
throughout this paper. In short, subjects learn quickly to distin-
guish the specific context and predict the immediate appearance
of the syllables, as well as to retrieve practically without errors the
response syllables, even in those experiments that included
memory impairments.
Discussion
The main finding of this study is the
demonstration, in humans, that previ-
ously consolidated declarative memory
returns to a labile state and becomes sub-
ject to stabilization again. This process
of labilization-reconsolidation is trig-
gered by the presentation of a reminder
and characterized by the possibility that
a second training can impair the declara-
tive memory within a time window.
The demonstration backs the uni-
versality of the reconsolidation phe-
nomenon, which is consistent with the
idea that general principles of memory
organization, as well as basic compo-
nents of the mechanisms serving
memory, would be used across evolution
by phylogenetically very disparate ani-
mals (Carew 2000; Pedreira and Mal-
donado 2003). Moreover, the conclusion
that a consolidated declarative memory
can be actually disrupted after a reminder
has potential clinical relevance as has
been suggested several times (Bustos et al.
2006; Debiec et al. 2006; Foa 2006).
In order to analyze how the reminder triggers reconsolida-
tion, it seems pertinent to deal separately with the two mnemic
processes that are involved in the present study: the paired-
associate memory, and the one we term the prediction memory.
The reminder of the former memory consisted of the presenta-
tion of the L1 context (red light + New York City + jazz) and
immediately then, one cue syllable on the left side of the moni-
tors screen and the response box on the right. However, 2 sec
later the trial was abruptly interrupted. All the contextual param-
eters and the cue syllable vanished and only the notice on the
monitors screen announcing the unexpected end of the trial
remained. Thus, the reminder included also in this case what we
consider the double requirement for producing the memory la-
bilization and its following reconsolidation; namely, the initial
retrieval of the target memory by presenting the cue syllable and
the termination of the reminder without the appearance of the
correct response syllable (Pedreira et al. 2004). The memory re-
trieved by the cue syllable implies, as every memory, an expec-
tation (Dudai 2002), specifically, the subject here expects the
reinforcement of the correct response syllable. Therefore, the re-
minder closure marks the irreversible mismatch between what
was expected and what actually occurred. In this framework, we
discuss the possible functionality of the reconsolidation process
as an answer to such frustration. A wide range of memory flaws
could account for a failed prediction, ranging from outdated to
faulty or incomplete information. Therefore, it seems reasonable
to suppose (Nader et al. 2000b; Sara 2000b) that labilization-
reconsolidation plays a repair role by enabling the system to
integrate new information on the background of the past. This
memory updating mechanism would not entail an obligatory
phase of every retrieved memory but a mechanism of exception,
triggered only if the presentation of the unreinforced reminder
was terminated and if its duration was sufficiently short (Pedreira
et al. 2004). The demonstration that the process of memory re-
consolidation also occurs with declarative memory in humans
offers the remarkable opportunity of testing straightforwardly
the role of this process and its functional value. Experiments may
be performed aimed at testing that the reminder necessarily in-
cludes an episode of frustration, and also, that the updating of
Figure 4. The effect of an interfering training on memory reconsolidation lasts at least 48 h. (Ex-
periment 1) (Left) Groups (n = 10): controls A, L1, L2, TL1, TL2, Phase 1, and Phase 2, as in Figure 1;
5m, the 5 min between testings. (Right) Mean of total errors SEM on Day 4. **, P < 0.01; ***,
P < 0.001. (Experiment 2) (Left) Groups (n = 10): controls A, L1, L2, TL1, TL2, Phase 1, and Phase 2, as
in Figure 1; 5m, the 5 min between the reminder trial and L2-training, or for the 5 min between
testings; R, as in Figure 3. (Right) Mean of total errors SEM on Day 4. **, P < 0.01.
Memory reconsol i dati on i n humans
Learning & Memory 299
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information occurs during the reconsolidation period but not
when the memory is stable again. However, although results of
the present experiments as well as others performed with other
animals (Pedreira et al. 2002; Debiec and LeDoux 2004; Suzuki et
al. 2004) suggest that reconsolidation is engaged when unex-
pected events occur, it is pertinent to note that demonstrations
of reconsolidation have also been shown using the identical
stimuli as was employed during training (Duvarci and Nader
2004; Eisenberg and Dudai 2004).
The reconsolidation of the paired-associate memory was in-
terrupted by the learning of other verbal material (Mller and
Pilzecker 1900). The use of a training session as an amnesic agent,
instead of some invasive agents such as chemical blockers or
physical interferences, has the advantage of being free of unde-
sirable metabolic side effects or physical perturbations. More-
over, although both trainings were given to acquire different
verbal materials, they share several parameters of the experimen-
tal context as well as the pattern of syllables presentation, thus
involving clearly related memories. This gave the possibility of
disclosing retrieval interferences during testing and allowed to
use the effect of the RIF as a measure of a deficit in the target
memory. The RIF effect was detected in all the experiments of
this study except in those that included memory impairments.
The memory impairment is not disclosed by failures of its own
retrieval but the faultlessness in the retrieval of a related memory.
Thus, the absence or defect of RIF, from L1-retrieval on the later
L2-retrieval, proved to be an indicator of an impairment of the
paired-associate memory. A study that takes into account the
retrieval interaction and evaluates through them the deficits in
memory storage, seems to be more in keeping with the view that
memories are not stored in isolation of other memories but in-
tegrated into complex networks (Levy and Anderson 2002; Ber-
man et al. 2003; Debiec et al. 2006), a proposal especially valid in
the case of the human declarative memory. Thus, the RIF effect
has been used throughout this paper for testing long-term
memory and post-reactivation long-term memory, but if it is
used at a very short time after learning it will allow us to study
intriguing questions of the reconsolidation theory as to howlong
the memory remains labile but intact after reactivation (Pedreira
et al. 2004), and also to test the specificity of the RIF effect (Nader
et al. 2000a).
In contrast with what has just been shown, the association
between the specific context and the following display of the cue
syllables (the prediction memory) was never disrupted by the
second training. The prediction memory level was assessed by
the number of incorrect responses in pressing the expectancy
keys in all of the trials (actual and fake trials). The learning curves
fell abruptly, and subjects showed no mistakes after seven to
eight trials. At test sessions, all the subjects demonstrated near
100% of correct responses, and no statistical differences were
disclosed for any comparison between groups. Thus, the block-
ade of the consolidation (or reconsolidation) by a second train-
ing is found for paired-associate memory but not for the predic-
tion one. This contrast could be explained by the difference in
number of trials between prediction and paired-associate learn-
ing (10 and 32 trials, respectively), or by the difference in com-
plexity between them, which makes one memory weaker than
the other. Alternatively, another account may be offered, at least
concerning reconsolidation, based on the hypothesis that for this
to take place, the predicted reinforcement should not be deliv-
ered during the entire reminder presentation (Pedreira et al.
2004). According to the experimental protocol, however, the re-
minder trial ends after a 2-sec exposure of the cue syllable. Thus,
for the paired-associate memory, the promised reinforcement
failed to occur, while for the prediction memory, it was fulfilled.
Therefore, the same reminder fails to produce reconsolidation if
the recalled memory includes promises that are fulfilled, and
triggers reconsolidation if it includes predictions that fail.
At this juncture, we would like to note two new perspectives
opened by the present results. First, it is of great value from a
clinical point of view, that a traumatic or pathogenic memory
could be reactivated by selective reminders and disrupted not
only by a pharmacological agent but also by other learning (Bus-
tos et al. 2006; Debiec et al. 2006; Foa 2006). Second, the possi-
bility is very promising for future reconsolidation studies of using
the human declarative memory as a model to address conten-
tious or still conjectural topics about reconsolidation, such as its
functionality (Frenkel et al. 2005; Debiec et al. 2006) or its role in
updating information (Sara and Hars 2006). It is pertinent to add
that in a recent work, published when the present manuscript
was under revision, it was reported that human episodic memo-
ries can undergo reconsolidation (Hupbach et al. 2007); however,
the omission of the RIF analysis deprives this work of relevant
information that would allow discarding alternative explana-
tions for the reminder effect.
Figure 5. (A) Impact of the context phase on the paired-associated
learning. (Upper panel) Diagram of the training session. The context
group received the list of syllables after the presentation of the specific
context (5-sec red light, 5-sec red light plus New York City image, and 10
sec of light, image, and jazz music). The No-Context Group, was trained
in the same basic context (same dark room, personal computer, ear-
phones) but before the syllable presentation received a 20-sec interval
without any light, image, or sound stimulation. (Lower panel) Groups
(n = 10). Gray bar, the Context Group; white bar, the No-Context Group.
Mean of total errors SEM on testing, *, P < 0.05. (B) Training syllable
and prediction. Mean percentage errors SEM per trial during training.
Dotted line, syllable errors; solid line, prediction errors; black circles, ac-
tual trials; white circles, fake trials. The gray rectangle corresponds to the
training tail trails (last four syllable training trails and last 12 prediction
training trails). The training sessions (L1-training or L2-training) com-
prised 10 actual trials interspersed in 22 fake trials (total: 32 trials).
Forcato et al .
300 Learning & Memory
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Materials and Methods
Subjects
Two-hundred-eighty healthy undergraduate and graduate stu-
dents from Buenos Aires University volunteered for the study
(60% women, 40% men). Before their participation in the experi-
ment, subjects signed an informed consent approved by the
Comit independiente de tica para Protocolos de Investigacin
(CEPI) Hospital Italiano de Buenos Aires. Their ages ranged from
18 to 35 yr, with a mean of 25. The participants were randomly
assigned to one of 40 groups.
Procedure
Context formation and syllables presentation
Experiments took place in a dark roomand were conducted using
a personal computer. Each subject was provided with earphones,
and seated in front of a monitor placed ahead of a large screen on
the back wall (Fig. 6A). Each training trial included a first period
during which the context was formed (the context period), fol-
lowed by a second one during which a series of nonsense syl-
lables were presented as paired associates (the syllable period).
The context period comprised a fix sequence of three accumula-
tive steps: a first step of light alone projected on the large screen
for 5 sec; a second step of light plus image for 5 sec, with the
addition of an image displayed on the monitors screen; and a
third step of light plus image plus sound for 10 sec, with the
addition of a sound coming through the earphones. The specific
context persisted during the syllable presentation (Fig. 6B). The
syllable period that follows the former, started with the presen-
tation of a cue syllable on the left side of the monitors screen and
an empty response box on the right. Each cue syllable was taken
at random and successively from a list of five. Subjects were re-
quired to write down the corresponding response syllable before
5 sec. After finishing that period three situations were possible:
first, if no syllable was written, the correct one was shown for 4
sec; second, if an incorrect syllable was written, it was replaced by
the correct one and it was shown for 4 sec; and third, if the
correct response was given, it stayed for a further 4 sec. Immedi-
ately after, another cue syllable was shown and the process was
repeated until the list was over. All together, an actual trial lasted
65 sec (20 sec for the context period and 45 sec for the syllable
presentation).
The L1-training session
A trial of the List 1 (L1)-training had the three steps of the con-
text formation, each with two possible options: the light could be
red or blue; the image, a picture of Ravello or one of New York
City; the sound, a jazz song or an Italian one. Only one combi-
nation of these options (the specific context) was followed by the
syllables presentation of List 1. The trial that includes specific
context followed by the syllables presentation is termed the ac-
tual trial, while the others with only context (i.e., without syl-
lables presentation) are called the fake trials. In L1-training, the
specific context of the actual trial was: red light + picture of New
York City + jazz melody.
The L1-training consisted of 10 actual trials interspersed
with 22 fake trials (total: 32 trials), separated by a 3-sec intertrial
interval. List 1 was constituted by five pairs of nonsense cue-
response syllables in rioplatense Spanish: ITE-OBN, ASP-UOD,
FLI-AIO, NEB-FOT, COS-GLE (bold
type: cue syllable; regular type: response
syllable) (Fig. 6C). Subjects that failed to
reach 70% of correct responses dur-
ing the last four actual trials were ex-
cluded.
To evaluate the prediction memory
(the association between the specific
context and the syllable presentation),
subjects were instructed to press the YES
or NO button (the expectancy keys) on
the keyboard 3 sec after the last step of
the lightimagesound sequence has
started. The subject has to press YES if he
thinks that the syllable presentation is to
come about, NO in the opposite case.
The L2-training session
A trial of the L2-training had the three
steps of the context formation, each
with two possible options: the light
could be green or yellow; the image, a
picture of a forest or one of Waikiki
beach; the sound, a symphony or a blues
song. In the L2-training, the specific
context of the actual trial was: green
light + picture of the forest + symphony.
The L2-training consisted of 10 ac-
tual trials interspersed with 22 fake trials
(total: 32 trials), separated by a 3 sec in-
tertrial interval. List 2 was constituted by
five pairs of nonsense cue-response syl-
lables in rioplatense Spanish: OEN-SRO,
DRI-CRE, AIC-POA, TIU-PLA, KEC-
CLO (bold type: cue syllable; normal
type, response syllable) (Fig. 6C). Sub-
jects that failed to reach 70% of correct
responses during the last four actual tri-
als were excluded.
To evaluate the prediction memory,
subjects were instructed to press the YES
or NO button, as above.
Figure 6. Experimental protocol. Subjects were submitted to two different training sessions (L1-
training and L2-training), separated by a time interval, in a 3-d experiment. During each training trial,
the subject was seated in front of a monitor placed ahead of a large screen. (A) Diagram of the
experimental room. Symbols are explained in the right panel. (B) An actual trial included a sequence
of three accumulative steps termed the specific context: first step, light (color illumination of the large
screen); second step, image (a picture on the monitor); third step, sound (music melody from ear-
phones). Ten seconds after presentation of the sound, five pairs of cue-response syllables were pre-
sented successively and at random order (each pair consisted of two squares superimposed on the
monitors image: the left square was the cue box, which includes a nonsense syllable of three letters,
and the right one, the empty response box, was where the subject was required to write down the
corresponding syllable). Expectancy keys: Participants were told that as soon as the sound presentation
was started they should press one of the two keys. Yes, I expect the cue-response display comes:
No, I dont. (C) Lists: L1 and L2 differed each other in cue-response pairs as well as in color, light,
image, and sound that precede them or those of the respective fake trials.
Memory reconsol i dati on i n humans
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Testing
The testing session consisted of two retrievals of verbal material
presented in alternative order: TL1 corresponding to the first
training (L1) followed by TL2 corresponding to the second train-
ing (L2) (groups A) or TL2 followed by TL1 (groups B). Each
retrieval (TL1 or TL2) consisted of four actual trials of list L1 or
list L2, respectively, interspersed with eight of the respective fake
trials (total: 12 trials each). Thus, each retrieval had a distribution
in number and types of trials similar to the last 12 trials of the
respective training (termed the training tail). Subjects had to
press the key YES or NO (once the last step of the context period
was started).
Reminder
The reminder trial included the specific context of L1. Immedi-
ately after completing it, and as expected, a cue syllable appeared
on the left side of the monitors image and the response box on
the right. However, 2 sec later a notice displayed on the monitor
announced that the session has to being suspended, thus not
allowing the subject to write down the response syllable in the
box.
Demo
Before the training session, participants were confronted with a
demo program to receive all the instructions and to understand
the goal of the task. The program consisted of four trials, similar
in structure to but with a context and two pairs of nonsense
syllables different from those of L1- or L2-training.
Statistics
Results are reported as mean number of errors for each retrieval
during test session. Data from each experiment were first ana-
lyzed with a one-way ANOVA with a number of levels equal to
the number of retrievals. It was followed by a priori planned
comparisons (FISHER, = 0.05) between the number of errors of
L1-retrieval at phase 1 (or at phase2) of testing vs. the respective
control L1CTL; and L2-retrieval at phase 2 (or phase 1) of testing
vs. the respective control L2-CTL.
The choice of this statistical approach was on line with the
purpose of evaluating the mutual influence at testing between
the retrievals of two different verbal materials acquired under
different training conditions. Consequently, the effect was prop-
erly revealed by contrasting the performance for L1-retrieval of a
group that received both trainings with the performance of a
control group that received only L1, and separately, by contrast-
ing the performance for L2-retrieval of a group that received both
trainings with a group that received only L2.
For training curves the analysis was conducted by using the
method of successive regression (Lozada et al. 1990).
Acknowledgments
We thank our subjects for their cooperation, S. Maldonado for
advice in linguistic matter, M. Camorino for assistance in illumi-
nation, and A. Delorenzi and L.M. Prez-Cuesta for their helpful
comments on the manuscript. We also thank M.D. Trebucq for
assistance in preparation of the manuscript. This work was sup-
ported by FONCYT (Grant PICTR 00349), UBACYT (Grant X326),
and FUCIBA.
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Received November 22, 2006; accepted in revised form February 7, 2007.
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