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Intra-Administration Associations and Withdrawal Symptoms: Morphine-

Elicited Morphine Withdrawal

Article  in  Experimental and Clinical Psychopharmacology · March 2004

DOI: 10.1037/1064-1297.12.1.3 · Source: PubMed

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Experimental and Clinical Psychopharmacology Copyright 2004 by the American Psychological Association, Inc.
2004, Vol. 12, No. 1, 3–11 1064-1297/04/$12.00 DOI: 10.1037/1064-1297.12.1.3

Intra-Administration Associations and Withdrawal Symptoms:

Morphine-Elicited Morphine Withdrawal
Robert V. McDonald and Shepard Siegel
McMaster University

On the basis of a conditioning analysis, some drug “withdrawal symptoms” are conditional
responses elicited by stimuli paired with the drug effect. Prior demonstrations of conditional
elicitation of withdrawal symptoms evaluated the role of environmental cues; however,
pharmacological cues also typically signal a drug effect. Within each administration, early
drug onset cues (DOCs) may become associated with the later, larger drug effect (intra-
administration associations). This experiment evaluated the contribution of intra-administra-
tion associations to withdrawal symptoms. The results indicated that (a) 5 mg/kg morphine
elicited behavioral and thermic withdrawal symptoms in rats previously injected on a number
of occasions with 50 mg/kg morphine and that (b) DOC-elicited withdrawal symptoms are
not a sensitized response to the opiate but rather an associative phenomenon.

Events occurring during drug administration correspond
to a Pavlovian conditioning trial (e.g., Dworkin, 1993; Ram-
say & Woods, 1997; Siegel, Baptista, Kim, McDonald, &
Weise-Kelly, 2000). Cues accompanying the drug effect
function as conditional stimuli (CSs), and the direct drug
effect constitutes the unconditional stimulus (US). Prior to
any learning, this US elicits responses that compensate for
drug-induced disturbances. These responses that compen-
sate for the drug effect frequently act as unconditional
responses. After some pairings of the predrug CS and phar-
macological US, the drug-compensatory responses are elic-
ited as conditional responses (CRs). We (see Siegel et al.,
2000) and others (e.g., Poulos & Cappell, 1991; Ramsay &
Woods, 1997) have hypothesized that, when a drug is ad-
ministered in the context of the usual drug administration
cues, these drug-compensatory CRs contribute to tolerance;
that is, they attenuate the drug effect. Moreover, when the
subject is in the presence of the usual drug administration
cues but the drug is not administered, these CRs may be
expressed as “withdrawal symptoms.” On the basis of the
conditioning analysis of tolerance and withdrawal symp-
toms, then, these phenomena should be more pronounced in
the context of the usual drug administration cues than in the
context of alternative cues.
Most experiments designed to assess the role of predrug
cues in drug effects have assessed diffuse environmental
Robert V. McDonald and Shepard Siegel, Department of Psy-
chology, McMaster University, Hamilton, Ontario, Canada.
Robert V. McDonald is now at the Department of Psychology,
Wilfrid Laurier University, Waterloo, Ontario, Canada.
This research was supported by research grants from the Na-
tional Institute on Drug Abuse (DA11865) and the Natural Sci-
ences and Engineering Research Council of Canada (00298) to
Shepard Siegel.
Correspondence concerning this article should be addressed to
Shepard Siegel, Department of Psychology, McMaster University,
Hamilton, Ontario L8S 4K1, Canada. E-mail: siegel@mcmaster.ca
cues. Thus, rats are more tolerant to a drug when it is
administered in the usual drug-paired environment (e.g., the
room where the drug had been administered in the past) than
when it is administered elsewhere (e.g., in a different room;
see review by Siegel et al., 2000). Similarly, rats display
more behavioral withdrawal symptoms when, in the ab-
sence of the drug, they are placed in the drug-paired envi-
ronment than when they are placed in an alternative envi-
ronment (see review by Siegel & Ramos, 2002). Recently,
it has become apparent that a variety of interoceptive cues
may become associated with a drug and may control the
display of tolerance (see Siegel & Ramos, 2002). Of special
relevance to the present experiment are reports that a small
dose of a drug may serve as a CS, signaling a subsequent,
larger dose of the drug (Goddard, 1999). Such a pharmaco-
logical pairing procedure has been termed intradrug condi-
tioning (Siegel et al., 2000). Greeley, Lê, Poulos, and Cap-
pell (1984) provided the first demonstration of intradrug
conditioning. In this Greeley et al. study, some rats (paired
group) consistently were injected with a low dose of ethanol
(0.8 g/kg) 60 min prior to injection of a high dose of ethanol
(2.5 g/kg). Other rats (unpaired group) were injected with
the low and high doses on an unpaired basis. When tested
for the tolerance to the hypothermic effect of the high dose
following the low dose, paired-group rats, but not unpaired-
group rats, displayed tolerance. Moreover, if the high dose
of ethanol was not preceded by the low dose, paired-group
rats failed to display their usual tolerance. More recently, it
has been reported that an intradrug association contributes
to tolerance to morphine (Cepeda-Benito & Short, 1997;
Sokolowska, Siegel, & Kim, 2002).
Intradrug conditioning findings have important implica-
tions for an associative analysis of drug effects. A gradual
increase in systemic concentration is an inevitable conse-
quence of most drug administration procedures. That is,
even if there is no explicit attempt to pair administration of
a small drug dose with the subsequent administration of a
larger drug dose, such pairings may result when a drug is

3
4 MCDONALD AND SIEGEL
administered. Within each administration, small, drug onset
cues (DOCs) typically precede subsequent larger drug ef-
fects, and thus DOCs may serve as CSs. Siegel et al. (2000)
termed associations naturally occurring within a single ad-
ministration intra-administration associations to distinguish
them from the intradrug associations that develop when a
small drug dose is explicitly administered prior to a larger
drug dose. Several investigators have suggested that intra-
administration DOCs constitute an important component of
the CS that elicits the drug-compensatory CRs that mediate
tolerance (e.g., Grisel, Wiertelak, Watkins, & Maier, 1994;
Kim, Siegel, & Patenall, 1999; Poulos & Cappell, 1991;
Tiffany, Petrie, Baker, & Dahl, 1983; Walter & Riccio,
1983).
On the basis of the conditioning analysis, withdrawal
symptoms—a manifestation of a pharmacological CR—
should be elicited not only by drug-associated environmen-
tal cues but also by drug-associated pharmacological cues.
If an intra-administration association is formed during a
series of morphine administrations, presenting a small dose
of the opiate might be expected to reproduce the early effect
of the drug—an effect that has become associated with the
subsequent, larger effect. Typically, for the organism with a
history of morphine administration, administration of the
opiate prevents withdrawal symptoms from occurring. We
are suggesting that a small dose of the drug would actually
elicit the symptoms in organisms that previously have been
administered larger doses of the drug. In the present exper-
iment rats were repeatedly injected with a large dose of
morphine (50 mg/kg) prior to testing with a smaller dose (5
mg/kg). We expected that these rats would have acquired an
intra-administration association, and thus 5 mg/kg morphine
would function as a DOC and elicit withdrawal symptoms.
The purpose of this experiment was to evaluate this coun-
terintuitive prediction.
Evaluation of an associative analysis of DOC-elicited
withdrawal symptoms is complicated by the fact that the
intra-administration CS and US are intrinsic parts of the
same stimulus. Typical conditioning control procedures
(e.g., random CS–US presentations; see Rescorla, 1967) are
not feasible because the later, larger drug effect (US) cannot
be presented independently of DOCs (CS). Thus, following
administrations of a large drug dose, it is possible that
small-dose effects may represent a sensitized response
rather than a CR. For example, a small morphine dose might
elicit hyperactivity as a nonassociative, sensitized response
in morphine-experienced rats (e.g., Powell & Holtzman,
2001; Sokolowska et al., 2002), and behavioral displays of
apparent drug withdrawal symptoms might actually be be-
haviors secondary to this hyperactivity. Indeed, a recent
study using a methodology similar to that used in the
present study (i.e., small doses of heroin administered to
subjects with a history of exposure to substantially larger
doses of the drug) resulted in the observation of heroin-
elicited hyperalgesia, although this effect was attributed to
sensitization processes rather than to Pavlovian condition-
ing (Célèrier, Laulin, Corcuff, Le Moal, & Simonnet, 2001).
One technique we used in the present experiment to
distinguish between an associative and sensitization inter-
pretation of the results was to record the thermic as well as
the behavioral effects of morphine withdrawal. Administra-
tion of morphine, at the doses used in the conditioning phase
of the present study, elicits hyperthermia (e.g., Broadbent &
Cunningham, 1996; Clark, 1979; Eikelboom & Stewart,
1979), and sensitization to this hyperthermia may be seen
over the course of repeated morphine administrations; that
is, the morphine-elicited hyperthermia becomes more pro-
nounced (Broadbent & Cunningham, 1996). In contrast,
morphine withdrawal is accompanied by hypothermia. This
withdrawal hypothermia has been characterized as “one of
the most consistent and reliable parameters of [morphine]
withdrawal in the rat” (Gianuttsos, Drawbaugh, Hynes, &
Lal, 1975, p. 302). Thus, a finding that a small dose of
morphine elicits withdrawal-like hypothermia in rats with a
history of administration of larger doses of morphine (and
that are sensitized to the drug’s hyperthermic effect) would
suggest that the thermic response to the small dose is an
associative rather than a sensitized response.
A second technique we used to address the potential
confound of sensitization of morphine-induced behaviors
was to repeatedly administer the small dose of morphine in
rats conditioned with the large dose of morphine to evaluate
extinction of DOC-elicited withdrawal symptoms. If the
thermic and behavioral responses elicited by the small dose
represented a sensitized response, these responses would be
expected to increase over the course of repeated adminis-
tration of the opiate. However, if these small-dose-elicited
responses were CRs, such repeated administrations of the
putative CS would result in extinction and, therefore, a
decrease in both the thermic and behavioral indices of
withdrawal.
Method
Subjects
The subjects, 35 male Sprague–Dawley rats (Charles River, St.
Constant, Quebec, Canada), weighed 230 –285 g on the 1st day
of the experiment. They were individually housed in clear plastic
cages, with food and water available ad libitum except during a
10-min behavioral assessment period on test days. Subjects were
maintained on a 12-hr light– dark cycle (lights on at 0600). All
conditioning and testing took place during the light portion of
the cycle. Throughout the time of their participation in the exper-
iment, rats were housed in their homecages in the experimental
environment.
Surgery
One week prior to the start of conditioning, all rats were im-
planted with radiotelemetric temperature transmitters (model
PDT4000; Mini-Mitter, Bend, WA). These transmitters are en-
closed in a 22 mm ⫻ 8 mm capsule and weigh 1.6 g. Rats were
anesthetized with a 2:1:1 mixture of ketamine, xylazine, and
saline, which was injected intraperitoneally. A 1.5-cm incision was
made through the abdominal wall, and the transmitter capsule was
inserted into the abdominal cavity. Subjects received an oral an-
 MORPHINE-ELICITED MORPHINE WITHDRAWAL
tibiotic (Novo-Trimel, Toronto, Ontario, Canada) in their drinking
water during the 1-week recovery period following surgery.
Drugs and Apparatus
All solutions were injected intraperitoneally. Morphine sulfate
(British Drug House, Toronto, Ontario, Canada) was dissolved in
physiological saline such that the injection volume for both large
(50 mg/kg) and small (5 mg/kg) doses of the opiate was 1 ml/kg.
Saline was also injected at this volume.
Behavioral testing was conducted in one of six identical clear
acrylic observation chambers (30 cm ⫻ 30 cm ⫻ 30 cm) located
in the experimental room. These chambers rested on the telemetric
receiving units (model ER4000; Mini-Mitter, Bend, WA). Tem-
perature data were automatically collected by the controlling soft-
ware (VitalView, Mini-Mitter, Bend, WA) at 20-min intervals
during all conditioning and test trials.
Procedure
Prior to conditioning, all subjects were habituated to the testing
chambers during a single, 6-hr session. Following habituation,
subjects were randomly assigned to one of six groups: 50 –5,
50 – 0, 5–5, 5– 0, 0 –5, and 0 – 0 (10 rats were assigned to Group
50 –5, and 5 rats were assigned to each of the other groups). The
first number of the group designation indicates the treatment
during conditioning: 50 mg/kg morphine, 5 mg/kg morphine, or 0
mg/kg morphine (i.e., physiological saline). The second number
indicates the treatment at test: 5 mg/kg morphine or 0 mg/kg
morphine (saline). Conditioning began the day following habitu-
ation and consisted of intraperitoneal injection of 50 mg/kg mor-
phine (Groups 50 –5 and 50 – 0), 5 mg/kg morphine (Groups 5–5
and 5– 0), or saline (Groups 0 –5 and 0 – 0) once daily for 10
consecutive days. On the 1st conditioning day, each rat’s cage was
placed on a receiving pad. Temperature data were recorded for 30
min prior to injection, after which subjects were removed from
their homecage, injected, and immediately returned to the
homecage. Temperature data were recorded for 260 min following
injection. The time of the daily injections was varied from 1000 to
1600 in an irregular order. Temperature data were converted to
difference scores relative to a baseline temperature measure. This
baseline measure was obtained by calculating the average of the
last two preinjection temperature measurements.
The day following the 10th conditioning day was the 1st test
day. Subjects were individually removed from their homecages
and placed in the observation chambers, which replaced the
homecages on the receiving pads. After 30 min of temperature data
collection, subjects were injected either with 5 mg/kg morphine
(Groups 50 –5, 0 –5, and 5–5) or saline (Groups 50 – 0, 0 – 0, and
5– 0) and were videotaped for a period of 10 min following
injection (temperature data collection continued throughout behav-
ioral testing). At the end of the behavioral data collection period,
subjects were transferred back to the homecage and temperature
data collection continued for the rest of the 250-min postinjection
interval. All subjects in Group 50 –5 were given additional test
trials for 4 more days to assess the effects of repeated administra-
tion of the small dose of morphine. Videotaped behaviors were
later analyzed using behavioral data collection software (The Ob-
server, Noldus Information Technology, Wageningen, the Nether-
lands). The behaviors scored during testing were rearing, wet dog
shakes, mouth movements, ear wipes, and genital licks (see Mac-
Rae & Siegel, 1997; McDonald & Siegel, 1998). Scoring was blind
(i.e., the rater had no knowledge of the rats’ group assignments).
5
Results
Temperature Data
Conditioning Phase
Rats in the two groups trained with the large dose of
morphine (Groups 50 –5 and 50 – 0) were treated identically
during the conditioning phase of the experiment, each being
injected daily with 50 mg/kg morphine. Similarly, rats as-
signed to Groups 5–5 and 5– 0 were injected daily with 5
mg/kg morphine during conditioning, and rats assigned to
Groups 0 –5 and 0 – 0 were injected daily with saline during
conditioning. Figure 1 displays the mean difference from
baseline temperature (⫾1 SEM) for groups injected with
each substance on the 1st day of conditioning and the final
(10th) day of conditioning.
As can be seen in Figure 1, administration of morphine on
Conditioning Day 1, at a dose of either 50 or 5 mg/kg,
resulted in a hyperthermic response, with the magnitude of
the hyperthermia being similar for both doses. Administra-
tion of saline resulted in a less extreme and much briefer
hyperthermic response. The hyperthermia seen in saline-
injected rats likely results from the stress induced by the
handling and injection procedures (Broadbent & Cunning-
ham, 1996). As can be seen by comparing Conditioning
Day 10 with Conditioning Day 1, the magnitude of mor-
phine-elicited hyperthermia increased over this phase of the
experiment for both drug-injected groups, and by the final
day of conditioning, the rats injected with the higher dose of
the opiate displayed more pronounced hyperthermia than
the rats injected with the lower dose of the opiate. In
contrast with the effects of repeated morphine administra-
tion, the stress-elicited hyperthermia elicited by repeated
saline injections was less pronounced on the final condi-
tioning day than on the initial conditioning day.
These observations were confirmed by a mixed-design
analysis of variance (ANOVA) of the data summarized in
Figure 1. The three-way interaction between group, condi-
tioning day, and time within each day was statistically
significant, F(24, 384) ⫽ 3.38, p ⬍ .001. Subsequent lower-
order ANOVAs revealed the sources of the interaction.
Within-group comparisons indicated that subjects condi-
tioned with either 50 or 5 mg/kg morphine displayed greater
hyperthermia on Day 10 than on Day 1, F(1, 14) ⫽ 6.75,
p ⫽ .02, and F(1, 9) ⫽ 6.00, p ⬍ .04, respectively. The
altered effect of saline injection on body temperature from
Conditioning Day 1 to Conditioning Day 10 was manifest as
a significant interaction between conditioning day and time
postinjection for saline-injected rats. The source of the
interaction is apparent in Figure 1; the brief hyperthermia
seen in saline-injected rats lasted less than 2 hr and was less
pronounced on the final conditioning day than it was on the
1st conditioning day, F(1, 9) ⫽ 9.92, p ⫽ .01, for Postin-
jection Minutes 10 –110.
Between-group comparisons indicated that, on Condi-
tioning Day 1, the effect of group was significant, F(2,
32) ⫽ 25.2, p ⬍ .001. Although rats conditioned with
either 50 or 5 mg/kg morphine were hyperthermic on this
6 MCDONALD AND SIEGEL
Figure 1. Mean differences from baseline temperatures (⫾1 SEM) on Conditioning Days 1 and 10
for groups injected with 50 mg/kg morphine, 5 mg/kg morphine, or physiological saline on each
conditioning day.
1st conditioning day, compared with saline controls (all
ps ⬍ .001), the difference between the two morphine-
injected groups was not significant. The effect of group was
also significant on Conditioning Day 10, F(2, 32) ⫽ 164,
p ⬍ .001; however, in contrast with the 1st conditioning
day, all pairwise comparisons between groups were signif-
icant (all ps ⱕ .003). That is, in morphine-experienced rats,
the drug elicited a greater temperature elevation in rats
repeatedly injected with 50 mg/kg than in rats repeatedly
injected with 5 mg/kg on the final conditioning day.
Test Phase
Test Day 1. On the test day, three groups of rats were
injected with 5 mg/kg morphine (Groups 50 –5, 5–5, and
0 –5) and three groups were injected with saline (Groups
50 – 0, 5– 0, and 0 – 0). There was no appreciable difference
in test session temperatures among the three saline-injected
groups and, for simplicity in data presentation, they are
collapsed into a combined saline control group. The mean
differences from baseline temperature (⫾1 SEM) for all
groups are shown in Figure 2. As is apparent in Figure 2,
rats tested with saline displayed minimal temperature alter-
ations. In contrast, rats conditioned with saline and injected
with morphine for the first time on this test session (Group
0 –5) displayed hyperthermia. Rats conditioned with 5
mg/kg morphine and injected with this dose of the opiate for
the 11th time on this test session (Group 5–5) displayed
even more pronounced hyperthermia than did Group 0 –5
rats, again demonstrating sensitization to the thermic effect
of morphine. In contrast, rats conditioned with 50 mg/kg
morphine and tested with 5 mg/kg morphine (Group 50 –5)
displayed a relatively brief hyperthermic response (com-
pared with rats in the other groups tested with 5 mg/kg
morphine). In fact, these Group 50 –5 rats became hypo-
thermic over the course of the test session.
The different time– effect curves for the groups were
confirmed by the results of a mixed-design ANOVA of the
data summarized in Figure 2, indicating a significant inter-
action between group and time, F(36, 372) ⫽ 19.7, p ⬍
.001. Of special relevance to the hypothesis evaluated in this
experiment, rats in Group 50 –5 displayed lower tempera-
tures than did rats in any other group (including the
saline-tested rats) at Postinjection Minutes 190 –230. A
mixed-design ANOVA of only these three intervals in-
dicated a significant group effect, F(3, 31) ⫽ 24.0, p ⬍
.001. Pairwise comparisons indicated that Group 50 –5
rats were significantly hypothermic compared with rats in
each of the other three groups, including saline-tested
rats (all ps ⱕ .005).
Extinction. Subjects in Group 50 –5, conditioned
with 50 mg/kg, were administered 5 mg/kg once daily for an
additional 4 days following the first test. Hypothermia was
assessed by computing, for each subject, the maximum
hypothermic response elicited by 5 mg/kg morphine, that is,
the maximum decrease in temperature from baseline that
occurred during the trial. On the first test trial, the mean
maximum hypothermic response (⫾1 SEM) observed in
these Group 50 –5 rats was ⫺0.88 °C (⫾0.17°). A single
sample t test revealed that the mean maximum hypothermia
seen in these subjects was significantly different from zero,
t(9) ⫽ ⫺5.22, p ⬍ .001. By the final (fifth) test session the
mean maximal hypothermic response was ⫺0.05 °C
(⫾0.18°) and was no longer significantly different from
zero. Each of the 10 rats in Group 50 –5 displayed a lower
temperature in response to 5 mg/kg morphine on the first
test session than on the final test session.
 MORPHINE-ELICITED MORPHINE WITHDRAWAL 7

Figure 2. Mean differences from baseline temperatures (⫾1 SEM) on the 1st test day for rats in
the combined saline-tested control group and for rats administered 5 mg/kg morphine following
conditioning with 50 mg/kg morphine (Group 50 –5), 5 mg/kg morphine (Group 5–5), or saline
(Group 0 –5).

Behavioral Data
Behavioral indices of withdrawal were collected on the
postconditioning test days. As is the case with the temper-
ature data, the results for the three saline-tested groups
(Groups 50 – 0, 5– 0, and 0 – 0) are collapsed into a com-
bined saline-tested control group for simplicity in data
presentation.
Test Day 1
Behavioral data collected on the 1st test day following 10
days of conditioning are summarized in Figure 3. As is
apparent in Figure 3A there were no significant differences
among groups in the mean frequency of rearing. There
were, however, significant differences among groups in
mean frequency of each of the other withdrawal symptoms:
wet dog shakes (see Figure 3B; F[3, 31] ⫽ 8.1, p ⬍ .001),
genital licks (see Figure 3C; F[3, 31] ⫽ 10.3, p ⬍ .001), ear
wipes (see Figure 3D; F[3, 31] ⫽ 10.7, p ⬍ .001), and
mouth movements (see Figure 3E; F[3, 31] ⫽ 19.0, p ⬍
.001). Examination of Figures 3B–E indicates that, as hy-
pothesized, the groups effects resulted because the small
dose of morphine elicited more withdrawal symptoms in the
group trained with the larger dose of the drug (Group 50 –5)
than it did in any other group. Pairwise comparisons (LSD)
indicated that, for each of the indices of morphine with-
drawal summarized in Figures 3B–E, the differences be-
tween Group 50 –5 and each of the other groups was statis-
tically significant (all ps ⬍ .001). None of the other pairwise
comparisons, on any withdrawal measure, were statistically
significant.
Extinction
Subjects in Group 50 –5 were administered 5 mg/kg for
an additional 4 days following the initial test. The mean
frequencies of each of the recorded behaviors (⫾1 SEM) on
alternate test days (Test Days 1, 3, and 5) are displayed in
Figure 4. As can be seen in this figure, the mean frequency
of each behavior decreased across days. In the case of
rearing (see Figure 4A), this difference was not significant.
Repeated measures ANOVAs of the other withdrawal
symptoms indicated that, for each of them, the effect of
extinction days was statistically significant, all Fs(2,
18) ⱖ 3.90, all ps ⬍ .04.
Discussion
Both the temperature and behavioral data indicate that
morphine withdrawal symptoms can be elicited by admin-
istration of morphine. Group 50 –5 rats, conditioned with 50
mg/kg morphine and tested with 5 mg/kg of morphine,
displayed withdrawal hypothermia and behavioral indices
of morphine withdrawal. The withdrawal symptoms elicited
by the 5 mg/kg dose in Group 50 –5 subjects were not
observed in other rats receiving the same dose at test
(Groups 5–5 and 0 –5), suggesting that such thermic and
8 MCDONALD AND SIEGEL

Figure 3. Mean frequency (⫾1 SEM) of rears (A), wet dog shakes (B), genital licks (C), ear wipes
(D), and mouth movements (E) for 10 min following injection on the 1st test day for rats in the
combined saline-tested control group and for rats administered 5 mg/kg morphine following
conditioning with 50 mg/kg morphine (Group 50 –5), 5 mg/kg morphine (Group 5–5), or saline
(Group 0 –5).

behavioral responses are not unconditional effects of 5
mg/kg morphine. Rather, this dose elicits such behaviors
only in rats with a history of substantially higher doses of
morphine.
Temperature was monitored during both the conditioning
and test phases of the experiment. Morphine elicited an
elevation in temperature, and sensitization developed to this
morphine-induced hyperthermia during the conditioning
phase. Moreover, this sensitization was more pronounced in
rats conditioned with the higher dose of morphine. During
the test session, rats administered 5 mg/kg morphine
(Groups 5–5, 0 –5, and 50 –5) initially displayed an uncon-
 MORPHINE-ELICITED MORPHINE WITHDRAWAL 9

Figure 4. Mean frequency (⫾1 SEM) of rears (A), wet dog shakes (B), genital licks (C), ear wipes
(D), and mouth movements (E) for 10 min following injection on the 1st, 3rd, and 5th test days for
rats in Group 50 –5.

ditional hyperthermic response, but this temperature eleva-
tion was less pronounced in Group 50 –5 rats than in rats in
the other groups tested with 5 mg/kg morphine. In fact, only
Group 50 –5 rats became hypothermic over the course of the
test session—a symptom of morphine withdrawal (e.g.,
Gianuttsos et al., 1975). The pattern of results is consistent
with the suggestion that 5 mg/kg morphine simulates DOCs
in rats previously injected with 50 mg/kg. Inasmuch as this
hypothermic effect of 5 mg/kg morphine seen in Group
50 –5 rats is opposite to the sensitized response, it is un-
likely that this response represents a sensitized response.
Rather, as expected on the basis of an intra-administration
analysis, the response to the small dose of morphine in rats
conditioned with a large dose is associative; the small dose
of the drug reproduces the early drug effects, DOCs, that
precede the full effects of 50 mg/kg morphine.
The hypothermic effect of 5 mg/kg morphine in Group
50 –5 subjects decreased over the course of repeated injec-
tions of the low dose of the drug, providing further evidence
that the morphine-elicited hypothermia is a CR. That is,
such a decrease would be expected if the association be-
tween DOCs and the later, larger drug effect (established
during conditioning) extinguished during testing. Such a
diminution in the thermic effect of the drug during testing
would not be expected if the response to 5 mg/kg morphine
was a nonassociative sensitized response.
The behavioral data are consistent with the temperature
data. Rats conditioned with 50 mg/kg and tested with 5
mg/kg (Group 50 –5) showed a greater frequency of all
withdrawal measures—with the exception of rearing—than
did subjects in other groups. Although there was no differ-
ence between groups in the frequency of rearing, it should
10 MCDONALD AND SIEGEL
be noted that there is some controversy concerning the
efficacy of rearing as a measure of morphine withdrawal.
There are studies of associative contributions to morphine
withdrawal that have used this rearing measure (e.g., Azor-
losa, Hartley, & Deffner-Rappold, 1994; Deffner-Rappold,
Azorlosa, & Baker, 1996; Kelsey, Aranow, & Matthews,
1990); however McDonald and Siegel (1998) provided ev-
idence suggesting that rearing in morphine-experienced rats
is a manifestation of novelty-elicited exploration rather than
morphine withdrawal (but see Azorlosa & Simmons, 1999).
All monitored withdrawal symptoms decreased over the
course of repeated presentation of 5 mg/kg morphine in
Group 50 –5 rats (with this effect being significant for all but
the rearing measure), suggesting that these behavioral re-
sponses to the small dose of morphine were associative.
That is, such a decrease would be expected if the association
between DOCs and the later, larger drug effect (established
during conditioning) extinguished during testing. Such a
diminution in the behavioral effects of morphine during
testing would not be expected if the response to the mor-
phine-elicited behaviors was a nonassociative sensitized
response.
The present demonstration that DOCs elicit withdrawal
symptoms is consistent with clinical observations. It is well
established that drug withdrawal symptoms and relapse to
drug use sometimes are precipitated by exposure to small
drug doses. Although there are various interpretations of
such priming effects (Shaham, Rodaros, & Stewart, 1994),
intra-administration associations may be responsible for
some instances of the phenomenon (Siegel & Ramos, 2002).
For example, it frequently has been reported that a small
dose of alcohol augments the craving for additional alcohol
and enhances subsequent alcohol consumption (see Anon-
ymous, 1939; Goddard, 1999). This small dose effect may
be attributable to the alcoholic’s association of the initial
effect of alcohol (i.e., DOCs experienced soon after inges-
tion of alcohol) with subsequent larger amounts of the drug:
“The signal value of a small drug dose may make a contri-
bution to ‘binge’ drinking and drug ‘priming’ effects in
humans” (Goddard, 1999, p. 418).
Not only is there the potential for DOCs to become
associated with later drug effects whenever a drug is ad-
ministered, but these pharmacological CSs are very salient
and thus may overshadow (see Kamin, 1969) simulta-
neously present exteroceptive cues (e.g., Goudie, 1990; Kim
et al., 1999; Walter & Riccio, 1983). There is evidence that
overshadowing occurs in pharmacological conditioning
(Walter & Riccio, 1983) and that DOCs overshadow simul-
taneously present environmental cues (Kim et al, 1999).
Such findings are relevant for interpreting some apparently
conflicting findings in the literature. Although there are
many reports that withdrawal symptoms are especially pro-
nounced in the presence of drug-paired environmental cues
(e.g., Azorlosa et al., 1994; Deffner-Rappold et al., 1996;
Kelsey et al., 1990), there are some reports to the contrary;
that is, it has been suggested that morphine withdrawal
symptoms are not more pronounced in the drug-paired
environment than in an alternative environment (Sobrero &
Bouton, 1989; Zellner, Dacanay, & Riley, 1984). Such
findings typically have been interpreted as evidence con-
trary to an associative analysis of withdrawal symptoms:
“These results [transenvironmental withdrawal symptoms]
lead us to suggest that withdrawal responses are not com-
pensatory conditioned responses elicited by drug-associated
cues” (Zellner et al., 1984, p. 179). In contrast, the results of
the present experiment suggest that transenvironmental dis-
plays of withdrawal symptoms may occur because the rel-
evant CS, DOCs, are present in both the drug-paired and
alternative environments, and this interoceptive pharmaco-
logical CS overshadows the exteroceptive cues manipulated
by the experimenter.
As elaborated elsewhere (Siegel & Ramos, 2002;
Sokolowska et al., 2002), recognition that intra-administra-
tion associations contribute to drug withdrawal symptoms
has implications for conditioning-based treatments of drug
addiction. Some drug addiction treatment protocols incor-
porate procedures to extinguish the association between
predrug cues and the drug. Cue exposure treatments typi-
cally do not incorporate extinction of DOCs. As discussed
by several investigators (e.g., Cepeda-Benito & Short, 1997;
Siegel & Ramos, 2002), the effectiveness of such behavioral
treatments may be improved if small drug doses are in-
cluded in the cue exposure procedure. Indeed, some inves-
tigators have described successful cue exposure treatment
procedures for problem drinking that incorporate priming
doses of alcohol (e.g., Sitharthan, Sitharthan, Hough, &
Kavanagh, 1997; Sitharthan, Sitharthan, & Kavanagh,
2001).
References
Anonymous. (1939). Alcoholics Anonymous: The story of how
many thousands of men and women have recovered from alco-
holism. New York: Works.
Azorlosa, J. L., Hartley, N. E., & Deffner-Rappold, C. (1994).
Context-specific morphine tolerance and withdrawal: The ef-
fects of interdose interval. Psychobiology, 22, 304 –311.
Azorlosa, J. L., & Simmons, E. L. (1999). Context-specific mor-
phine withdrawal: Evidence that rearing reflects withdrawal, not
exploration. Psychobiology, 27, 557–560.
Broadbent, J., & Cunningham, C. L. (1996). Pavlovian condition-
ing of morphine hyperthermia: Assessment of the interstimulus
interval and CS–US overlap. Psychopharmacology, 126, 156 –
164.
Célèrier, E., Laulin, J. -P., Corcuff, J. -B., Le Moal, M., &
Simonnet, G. (2001). Progressive enhancement of delayed hy-
peralgesia induced by repeated heroin administration: A sensi-
tization process. Journal of Neuroscience, 21, 4074 – 4080.
Cepeda-Benito, A., & Short, P. (1997). Morphine’s interoceptive
stimuli as cues for the development of associative morphine
tolerance in the rat. Psychobiology, 25, 236 –240.
Clark, W. G. (1979). Influence of opioids on central thermoregu-
latory mechanisms. Pharmacology Biochemistry and Behav-
ior, 10, 609 – 613.
Deffner-Rappold, C., Azorlosa, J. L., & Baker, J. D. (1996).
Acquisition and extinction of context-specific morphine with-
drawal. Psychobiology, 24, 219 –226.
Dworkin, B. R. (1993). Learning and physiological regulation.
Chicago: University of Chicago Press.
 MORPHINE-ELICITED MORPHINE WITHDRAWAL
Eikelboom, R., & Stewart, J. (1979). Conditioned temperature
effects using morphine as the unconditioned stimulus. Psycho-
pharmacology, 61, 31–38.
Gianuttsos, G., Drawbaugh, R., Hynes, M., & Lal, H. (1975). The
narcotic withdrawal syndrome in the rat. In S. Ehrenpreis & A.
Neidle (Eds.), Methods in narcotics research (pp. 293–309).
New York: Marcel Dekker.
Goddard, M. J. (1999). The role of US signal value in contingency,
drug conditioning, and learned helplessness. Psychonomic Bul-
letin and Review, 6, 412– 423.
Goudie, A. J. (1990). Conditioned opponent processes in the
development of tolerance to psychoactive drugs. Progress in
Neuro-Psychopharmacology and Biological Psychiatry, 14,
675– 688.
Greeley, J., Lê, D. A., Poulos, C. X., & Cappell, H. (1984).
Alcohol is an effective cue in the conditional control of toler-
ance to alcohol. Psychopharmacology, 83, 159 –162.
Grisel, J. E., Wiertelak, E. P., Watkins, L. R., & Maier, S. F.
(1994). Route of morphine administration modulates condi-
tioned analgesic tolerance and hyperalgesia. Pharmacology Bio-
chemistry and Behavior, 49, 1029 –1035.
Kamin, L. J. (1969). Predictability, surprise, attention, and condi-
tioning. In B. A. Campbell & R. M. Church (Eds.), Punishment
and aversive behavior (pp. 279 –296). New York: Appleton-
Century-Crofts.
Kelsey, J. E., Aranow, J. S., & Matthews, R. T. (1990). Context-
specific morphine withdrawal in rats: Duration and effects of
clonidine. Behavioral Neuroscience, 104, 704 –710.
Kim, J. A., Siegel, S., & Patenall, V. R. A. (1999). Drug-onset cues
as signals: Intraadministration associations and tolerance. Jour-
nal of Experimental Psychology: Animal Behavior Pro-
cesses, 25, 491–504.
MacRae, J. R., & Siegel, S. (1997). The role of self-administration
in morphine withdrawal in rats. Psychobiology, 25, 77– 82.
McDonald, R. V., & Siegel, S. (1998). Environmental control of
morphine withdrawal: Context specificity or stimulus novelty?
Psychobiology, 26, 53–56.
Poulos, C. X., & Cappell, H. (1991). Homeostatic theory of drug
tolerance: A general model of physiological adaptations. Psy-
chological Review, 98, 390 – 408.
Powell K. R., & Holtzman S. G. (2001). Parametric evaluation of
the development of sensitization to the effects of morphine on
locomotor activity. Drug and Alcohol Dependence, 62, 83–90.
Ramsay, D. S., & Woods, S. C. (1997). Biological consequences
of drug administration: Implications for acute and chronic tol-
erance. Psychological Review, 104, 170 –193.
View publication stats
11
Rescorla, R. A. (1967). Pavlovian conditioning and its proper
control procedures. Psychological Review, 74, 71– 80.
Shaham, Y., Rodaros, D., & Stewart, J. (1994). Reinstatement of
heroin-reinforced behavior following long-term extinction: Im-
plications for the treatment of relapse of drug taking. Behav-
ioural Pharmacology, 5, 360 –364.
Siegel, S., Baptista, M. A. S., Kim, J. A., McDonald, R. V., &
Weise-Kelly, L. A. (2000). Pavlovian psychopharmacology:
The associative basis of tolerance. Experimental and Clinical
Therapeutics, 8, 276 –293.
Siegel, S., & Ramos, B. C. (2002). Applying laboratory research:
Drug anticipation and the treatment of drug addiction. Experi-
mental and Clinical Psychopharmacology, 10, 162–183.
Sitharthan, T., Sitharthan, G., Hough, M. J., & Kavanagh, D. J.
(1997). Cue exposure in moderation drinking: A comparison
with cognitive– behavior therapy. Journal of Consulting and
Clinical Psychology, 65, 878 – 882.
Sitharthan, T., Sitharthan, G., & Kavanagh, D. J. (2001). Emo-
tional cue exposure for alcohol abuse: Development of a new
procedure to train moderation drinking in the context of dys-
phoria. Clinical Psychology and Psychotherapy, 8, 73–78.
Sobrero, A. P., & Bouton, M. E. (1989). Effects of stimuli present
during oral morphine administration on withdrawal and subse-
quent consumption. Psychobiology, 17, 179 –190.
Sokolowska, M., Siegel, S., & Kim, J. A. (2002). Intraadministra-
tion associations: Conditional hyperalgesia elicited by morphine
onset cues. Journal of Experimental Psychology: Animal Be-
havior Processes, 28, 309 –320.
Tiffany, S. T., Petrie, E. C., Baker, T. B., & Dahl, J. L. (1983).
Conditioned morphine tolerance in the rat: Absence of a com-
pensatory response and cross-tolerance with stress. Behavioral
Neuroscience, 97, 335–353.
Walter, T. A., & Riccio, D. C. (1983). Overshadowing effects in
the stimulus control of morphine analgesic tolerance. Behav-
ioral Neuroscience, 97, 658 – 662.
Zellner, D. A., Dacanay, R. J., & Riley, A. L. (1984) Opiate
withdrawal: The result of conditioning or physiological mech-
anisms? Pharmacology Biochemistry and Behavior, 20, 175–
180.
Received September 13, 2002
Revision received April 30, 2003
Accepted May 5, 2003 䡲