NEUROBIOLOGY OF LEARNING AND MEMORY 68, 189–196 (1997)

ARTICLE NO. NL973773

BRIEF REPORT
The NMDA-Receptor Antagonist Ketamine
as an Unconditioned Stimulus in
Taste Aversion Learning
Luis Aguado, Rafael del Valle, and Leticia Pérez
Facultad de PsicologıB a, Universidad Complutense, Campus de Somosaguas,
28023, Madrid, Spain

Three experiments studied the effectiveness of ketamine acting as an aversive uncon-

ditioned stimulus (UCS) in a conditioned flavor aversion procedure. In Experiment 1a,
three conditioning trials where sucrose was paired with ketamine produced a weak but
significant aversion to sucrose; Experiment 1b showed that this effect was not due to
a reduced consumption of sucrose caused by ketamine-induced neophobia. In Experi-
ment 2, acquisition of an aversion to sucrose paired with lithium chloride (LiCl) injec-
tions was retarded by prior repeated exposure to LiCl but not to ketamine. These
results are not consistent with an interpretation of previous results, showing that
ketamine impairs the acquisition of flavor aversions based on LiCl-induced illness, as
an example of the UCS preexposure effect. q 1997 Academic Press

In a report by Welzl, Alessandri, & Bättig (1990), the NMDA-receptor antag-

onist ketamine, injected prior to the conditioning trial, impaired conditioning
of an aversion to a flavor paired with illness induced by lithium chloride (LiCl).
This result was interpreted as showing that ketamine blocks the formation of
gustatory memory traces due to interference with long-term potentiation, a
plasticity mechanism observed in the vertebrate brain that has been related
to certain forms of learning and memory (e.g., Bliss & Collingridge, 1993).
However, Aguado, San Antonio, Pérez, Del Valle, & Gómez (1994) later re-
ported results that questioned this explanation. Specifically, these authors
showed that ketamine produces significant state-dependent learning effects
both in conditioning of a taste aversion (Experiment 1) and in latent inhibition
of the to-be-conditioned flavor (Experiment 3); that is, the apparent effects on
learning observed in ketamine-pretreated animals were significantly attenu-
ated when both learning and testing were carried out under the effects of
ketamine. Moreover, Aguado et al. showed that animals injected with ketamine
before each of three conditioning trials eventually reached a level of aversion
equivalent to that of control animals (Experiment 4). Although Aguado et al.
proposed an explanation for this pattern of results in terms of a reduction of
the salience of the taste stimulus by ketamine, they pointed out that the

Correspondence and reprint requests should be addressed to Luis Aguado, E-mail: psbas17@
sis.ucm.es.

189 1074-7427/97 $25.00

Copyright q 1997 by Academic Press
All rights of reproduction in any form reserved.

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190 AGUADO, DEL VALLE, AND PÉREZ

specific effect of a retardation of flavor aversion learning by ketamine might

alternatively be explained as an example of the well-known unconditioned
stimulus (UCS) preexposure effect (e.g., Cannon, Berman, Baker, & Atkinson,
1975). This effect consists of an attenuation of flavor aversion learning by prior
exposure to the UCS. This explanation is based on two main assumptions: the
first is that ketamine can act as an aversive UCS in taste aversion learning;
the second is that ketamine will produce a crossed preexposure effect with
LiCl, so that prior exposure to ketamine may impair later conditioning with
LiCl-induced illness as the UCS, based on possible common aversive effects of
both agents. The aim of this report is to test these specific assumptions, related
to the interpretation of the retardatory effects of ketamine on taste aversion
learning. To do so, in Experiments 1a and 1b ketamine was used as an UCS
to condition an aversion to sucrose; in Experiment 2, the ability of preexposure
to ketamine or LiCl to retard later conditioning of a LiCl-based aversion was
compared.
Experiment 1a evaluated the effectiveness of a 25-mg/kg dose of ketamine
injected after exposure to a sucrose solution to condition an aversion to sucrose.
This dose was chosen because it has been shown both by Aguado et al. (1994)
and by Welzl et al. (1990) to alter LiCl-based taste aversion learning when
given prior to the conditioning episode.
Sixteen experimentally naive male Wistar rats (Charles River), with weights
ranging from 290 to 320 g at the start of the experiment, were divided into two
groups of eight. They were kept in individual cages in a room with controlled
temperature and subjected to a 12 h–12 h light–dark cycle, with lights on at
8:00 AM. Throughout the experiment the animals had free access to food. Water
was restricted to two daily 30-min drinking periods. The experimental proce-
dures took place in the morning, during the light phase of the cycle, and were
always carried out in the home cage.
Three days before the start of the experiment, the animals were subjected
to a water deprivation schedule, consisting of two drinking periods, one in the
morning and another in the afternoon, where the animals had access to water
for 30 min. The afternoon drinking period was maintained throughout the
experiment. Conditioning trials consisted of a 30-min drinking period where
the animals had access for the first time to a 4% sucrose solution presented
in inverted centrifuge tubes provided with a metal spout. Immediately after
this, the animals of group KET were given an injection of ketamine, while
animals of group SAL were injected with saline, after which they were returned
to their home cages. There were three of these conditioning trials. The animals
had a recovery day after each injection day, where the animals had the same
drinking schedule as that in the deprivation days. Finally, three extinction
trials were given on consecutive days, where the animals had access to the
sucrose solution for periods of 15 min. During the conditioning trials consump-
tion of sucrose was limited to 20 ml, to reduce variability in the amount drunk,
which might have, itself, affected the strength of the aversion. However, during
test trials the animals were offered 30 ml of the solution, which virtually
amounts to unlimited access.
The results of Experiment 1a are presented in Fig. 1. Although both groups
consumed a fair amount of the sucrose solution during the conditioning and
extinction trials, it can be seen that in group KET this amount was reduced

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 KETAMINE AS AN AVERSIVE UCS 191

FIG. 1. Mean sucrose consumption over conditioning (cond) and extinction (ext) trials in Exper-
iment 1a. Group KET was injected with ketamine after drinking sucrose and group SAL was
injected with saline. Bars represent {SEM.

in the third conditioning trial and during the first and second extinction trials.
A significance level of p õ .05 was adopted for all statistical tests throughout
the paper. An ANOVA performed on consumption data in the conditioning and
extinction trials showed a significant effect of both group and trials [F(1, 14)
Å 6.797; F(5, 70) Å 4.709] and their interaction [F(5, 70) Å 5.527]. Analyses
of simple effects gave a significant effect of trials for both groups [F(5, 35) Å
6.79; F(5, 35) Å 3.18]. As to the group factor, significant differences were also
found between groups in the third conditioning trial and during the first and
second test trials. It should be recognized, however, that the significant differ-
ence obtained in the third conditioning trial was due not only to decreased
consumption in group KET, but also to increased consumption in group SAL.
But the significant effect of trials for both groups, together with the group 1
trial interaction, reflects the fact that consumption changed over trials in both
groups in opposite directions: a decrease in group KET due to the pairing of
sucrose with the effects of ketamine, and an increase in group SAL due to
habituation of neophobia to the flavor. Evidence that these results were not
due to differences in overall fluid intake between groups, based on hypodipsia
induced by ketamine, is provided by the consumption of water during the
afternoon drinking periods. In the trials where a significant suppression of
drinking was found in ketamine-injected animals, that is, in the third condi-
tioning trial and first and second extinction trials, water consumption was
12.13, 14.18, and 10.30 ml, respectively, for group KET, while for group CONT
the corresponding amounts were 10.27, 10.65, and 9.50 ml. There were no
between-group significant differences in water consumption in any of the trials.
Consequently, these results show that a dose of 25 mg/kg ketamine was effec-
tive enough to produce a mild but significant aversion to sucrose.
Decreased consumption in the KET group of Experiment 1 might possibly
be due to flavor neophobia induced by the drug, independent of its pairing

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192 AGUADO, DEL VALLE, AND PÉREZ

FIG. 2. Mean sucrose consumption over conditioning (cnd) and extinction (ext) trials in Experi-
ment 1b. Group KET-P received pairings of sucrose and ketamine, while group KET-NP received
unpaired presentations and group SAL was injected with saline. Bars represent {SEM.

with sucrose. In order to rule out this alternative, nonassociative explana-

tion, consumption in animals given sucrose – ketamine pairings should be
compared to that of animals given an equivalent amount of experience with
sucrose and ketamine, although in an unpaired manner. This was done in
Experiment 1b, which included three groups: group KET-P and group SAL,
receiving the same treatment as groups KET and CONT of Experiment 1,
respectively, and group KET-NP. This last group was equivalent to group
KET-P in terms of its experience with sucrose and ketamine, with the only
difference being that ketamine was injected 5 h after exposure to the flavor.
In Experiment 1b, 19 male Wistar rats, with weights ranging from 275 to
325 g at the start of the experiment, served as subjects. Fourteen animals,
divided in two groups of seven, were assigned to groups KET-P and KET-NP,
and five to group SAL. The animals were maintained under conditions similar
to those of Experiment 1.
All experimental treatments and parameters were similar to those of Experi-
ment 1a. The only exception was that in an attempt to observe a stronger
effect of conditioning, in this experiment animals received four, instead of
three, injections of ketamine. Thus, group KET-P had four conditioning trials
and group KET-NP four unpaired injections and exposures to sucrose. All
groups received three extinction trials after conditioning.
The results of this experiment are shown in Fig. 2. It can be seen that group
KET-P shows a steady reduction in consumption of the flavored solution over
trials, with maximum suppression of drinking by extinction trial 1, and that
consumption is recovered over extinction trials 2 and 3. Though suppression
of drinking was small and four conditioning trials were required to produce
an appreciable effect, both the difference with groups SAL and KET-NP and
the pattern of reduction and recovery of drinking over conditioning and extinc-
tion trials are suggestive of an acquisition process. However, far from showing

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 KETAMINE AS AN AVERSIVE UCS 193

a decrement in consumption, animals from group KET-NP increased their

consumption of sucrose over trials, reflecting habituation of neophobia to the
flavor. Although slightly weaker, this effect is also apparent in group SAL.
These impressions were confirmed by an ANOVA with groups and trials as
factors that gave a significant effect of group [F(2, 16) Å 8.81], trials [F(6, 96)
Å 6.34], and their interaction [F(12, 96) Å 2.29]. Analyses of simple effects
yielded a significant effect of trials in groups KET-P [F(6, 36) Å 3.29] and
KET-NP [F(6, 36) Å 7.07]. Finally, simple effects of group were significant at
all trials, except the first conditioning trial. Post-hoc analyses (Tukey’s test)
yielded significant differences between group KET-P and group KET-NP at all
those trials. Significant differences between group KET-P and group SAL were
also found on the third conditioning trial and the first extinction trial.
In conclusion, it can be said that suppression of drinking in group KET-P
was due to the pairing of sucrose with the effects of ketamine and not to
neophobia induced by ketamine. Thus, the results of this experiment replicate
the findings of Experiment 1 and allow us to rule out a nonassociative explana-
tion in terms of ketamine-induced neophobia.
In Experiment 2 we used a procedure explicitly designed for producing a
crossed preexposure effect, with ketamine as the preexposed agent and LiCl
as the UCS. To do so, animals were first given three injections of ketamine
and then had three conditioning trials where consumption of sucrose was
followed by a LiCl injection. A crossed preexposure effect might be expected
on the basis of the common ability of ketamine and LiCl to induce taste aver-
sions. If exposure to the aversive effects of the UCS is the requisite for showing
a preexposure effect on conditioning, this effect might then be found even when
these aversive effects are caused by different agents during preexposure and
during conditioning. Specifically, if ketamine is able to produce a crossed preex-
posure effect with LiCl, then animals pretreated with ketamine should show
a retardation of acquisition of a LiCl-based aversion. The design included two
additional groups. One group, injected with LiCl both during preexposure and
during conditioning, was included in order to compare the relative effectiveness
of LiCl and ketamine to generate the preexposure effect. A third nonprexposed
control group was also included.
Twenty male Wistar rats (Charles River) with weights ranging from 275
to 330 g at the start of the experiment were used as subjects. The animals
were kept under conditions similar to those of Experiment 1.
After a deprivation schedule similar to that used in Experiment 1, rats in
groups PRE-KET and PRE-LI received the preexposure treatment, consisting
of three injections of ketamine (25 mg/kg) or LiCl (.15 M, 10 ml/kg) given on
alternate days. Animals in group CONT received control injections of 1 ml
saline. There were seven animals in each of the preexposed groups and six in
the control group. Before each injection, the animals were given free access to
water for a period of 30 min. On the evening of every preexposure day the
animals had another 30-min drinking period and after each of these days a
recovery day was given, where animals had their usual two drinking periods.
Forty-eight hours after the third preexposure day all animals received the first
conditioning trial, consisting of access to 30 ml of a 4% sucrose solution for 30
min. There were three of these conditioning trials and, finally, a test trial
where the LiCl injection was omitted. These trials were separated by recovery

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194 AGUADO, DEL VALLE, AND PÉREZ

FIG. 3. Mean sucrose consumption over conditioning (cond) and test trials in Experiment 2.
Before conditioning, group PRE-KET was preexposed to ketamine and group PRE-LI to lithium
chloride, while group CONT was given saline injections. Bars represent {SEM.

days where the animals were allowed to drink water for the two usual 30-min
periods.
As can be seen in Fig. 3, all animals decreased their consumption of sucrose
during the conditioning trials, although this decrease was less pronounced in
group PRE-LI. Comparison of this group with group CONT shows the usual
LiCl preexposure effect. However, group PRE-KET did not show a weaker
aversion than group CONT and, if anything, these animals tended to drink
slightly less of the solution than the controls. Thus, preexposure to ketamine
was not effective in retarding the acquisition of a LiCl-based aversion. Statisti-
cal analyses showed significant effects of groups [F(2, 17) Å 4.681], trials [F(3,
51) Å 193.123], and their interaction [F(6, 51) Å 11.443]. Analyses of simple
effects revealed a significant effect of the group factor at all trials. Paired
comparisons confirmed the difference between group PRE-LI and groups PRE-
KET and CONT on the third conditioning trial and the first extinction trial.
The differences in consumption observed in the first conditioning trial might
simply reflect hypodipsia produced by repeated prior exposure to LiCl or keta-
mine. However, water consumption in the morning drinking period of the
previous recovery day, that is, the day between the last preexposure day and
the first conditioning day, was 11.65, 12.38, and 14.33 ml for groups PRE-
KET, PRE-LI, and CONT, respectively, and these means were not significantly
different. Alternatively, it might be that animals of both preexposed groups
manifested an increase of neophobia to the new sucrose flavor, due to their
prior experience with the aversive effects of lithium or ketamine (e.g., Domjan,
1977), though according to the results of Experiment 1b this possibility does
not seem plausible. In any case, differences in the acquisition rate cannot be
attributed to this differential consumption in the first trial. Although animals
of group PRE-KET drank less of the solution on this trial than controls, both

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 KETAMINE AS AN AVERSIVE UCS 195

groups acquired the aversion at a similar rate, the only retarded group being
that which was preexposed to LiCl.
The results presented in this report show that a dose of 25 mg/kg ketamine
acts as a mild aversive UCS when injected following consumption of the flavor
solution and that this effect is dependent on the pairing of the flavor with the
effects of the drug (Experiments 1a and 1b). Thus, ketamine shares with LiCl
its ability to induce conditioned taste aversions when injected after drinking a
new flavor. However, acquisition of an aversion to sucrose paired with lithium-
induced illness was not retarded by repeated exposure to the effects of keta-
mine before conditioning (Experiment 2). If the retarded acquisition found by
Aguado et al. (1994) in ketamine-injected animals was a case of a transient
UCS preexposure effect due to the injection of ketamine shortly before pairing
sucrose and LiCl, repeated exposure to ketamine before conditioning should
then have produced a durable, crossed preexposure effect with lithium. Usu-
ally, transient and durable preexposure effects are produced by the same agent,
the permanence of the effect depending on the number of preexposures and
the interval between preexposure and conditioning (e.g., Best, 1982). In our
case, an effect of preexposure to ketamine on the establishment of a LiCl-based
conditioned aversion would have been expected on the basis of the aversive
properties of ketamine shown in Experiment 1. The absence of this crossed
preexposure effect may be due to the mild aversive effects of ketamine or to
the different nature of the aversion induced by ketamine and LiCl. Of course,
although these results rule out one particular explanation of one of our previ-
ous findings, that is, the retardation of flavor aversion learning by ketamine,
they do not provide any direct evidence for the tentative explanation we offered
for the overall pattern of results reported by Aguado et al. (1994) in terms of
reduced salience of the flavor. We offered this last explanation as a more
parsimonious account, and one that is more consistent with our data, of the
effects of ketamine on the establishment of flavor memories than the one
offered by Welzl et al. (1990) in terms of interference with long-term potentia-
tion through its action as an NMDA-receptor antagonist. The possibility that
some behavioral effects of NMDA-receptor antagonists are caused by the un-
specific action of the compounds, and not to their alteration of memory mecha-
nisms, has been stressed repeatedly. Short-term effects of these compounds
on motor behavior, motivation, or perception have been pointed out repeatedly
(e.g., Keith & Rudy, 1990). Here we looked at a different kind of ‘‘unspecific,’’
long-term effect of NMDA-receptor antagonists, that is, their potential ability
to generate UCS preexposure effects that might retard subsequent learning.
The present results only exclude this effect and, in one particular case, the
acquisition of conditioned flavor aversions.

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