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Associative tolerance to nicotine analgesia in

the rat: Tail-flick and hot-plate tests
Article in Experimental and Clinical Psychopharmacology September 1998
Impact Factor: 2.71 DOI: 10.1037//1064-1297.6.3.248 Source: PubMed

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Copyright 1998 by the Am

Experimental and Clinical Psvchopliarniat-ology

1998, Vol. 6, No. 3, 248-254"

Associative Tolerance to Nicotine Analgesia in the Rat:

Tail-Flick and Hot-Plate Tests
Antonio Cepeda-Benito, Jose Reynoso, and Eric H. McDaniel
Texas A&M University
Previous assessments of associative nicotine tolerance may have confounded associative
effects with novelty-induced stress effects, instrumental learning effects, or both. That is,
subjects were tested in novel environments, allowed to practice the test response, or both
during the tolerance development phase. In the first study, 32 male Sprague-Dawley rats were
injected with various doses of nicotine and tested for nociception in the tail-flick and hot-plate
tests to assess nicotine's analgesic effects. In the second study, 35 rats received nicotine
explicitly paired or unpaired with a distinctive test context. All animals were equally
preexposed to the test environment, and none had the opportunity to practice the test response.
Paired rats developed greater nicotine tolerance than unpaired rats. This context-dependent
(associative) tolerance effect was found with both tail-flick and hot-plate tests.

Drug tolerance is a decrease in the effects of a drug dose

following repeated drug administrations. Drug tolerance is
central to the definition of drug dependence (American
Psychiatric Association, 1994), and several theorists assign
tolerance, or the mechanisms that produce tolerance, an
important role in the genesis and maintenance of addictive
behaviors (see Ramsay & Woods, 1997; Trujillo & Akil.
1995). For example, smokers may smoke more to compensate for the development of tolerance of the reinforcing
effects of nicotine. Likewise, tolerance of the ill effects of
nicotine may increase tobacco consumption by allowing
smokers to smoke more without feeling sick (Alexander &
Hadaway, 1982; Pomerleau, 1995; Trujillo & Akil). Thus,
nicotine tolerance can be a symptom of physical dependence
on nicotine (American Psychiatric Association, 1994) and a
contributor to the severity of the smoker's physical and
psychological dependence (Pomerleau).
There is considerable evidence that many examples of
drug tolerance represent the operation of classical conditioning (Young & Goudie, 1994), and some theorists have
proposed that "learning is the primary underlying cause"
(Ramsay & Woods, 1997, p. 170) of all drug tolerance
phenomena. Most of the support for the role of classical
conditioning in the development of drug tolerance comes
from investigations of tolerance to the analgesic effects of
morphine. In associative tolerance studies, animals receiving morphine explicitly paired with a distinctive test context

are more tolerant to morphine's analgesic effects than

animals receiving the same drug exposure regimen in an
environment other than the distinctive context (e.g., CepedaBenito & Tiffany, 1996a). The conditioning explanation for
these findings is that the distinctive context has become a
conditioned stimulus (CS) that elicits associative tolerance
effects (Baker & Tiffany, 1985; Poulos & Cappell, 1991;
Siegel, 1975). Some authors have also proposed that the
interoceptive effects of drugs may also function as CSs
capable of producing associative tolerance effects (e.g.,
Cepeda-Benito & Short, 1997; Grisel, Wiertelak, Watkins, &
Maier, 1994).
Despite the prominence of learning accounts of drug
tolerance, we know of only five studies that investigated the
impact of cue associations on the development of nicotine
tolerance (Caggiula et al, 1991, 1993; Caggiula, Epstein, &
Stiller, 1989; Epstein, Caggiula, Perkins, McKenzie, &
Smith, 1991; Epstein, Caggiula, & Stiller, 1989). These
studies investigated tolerance to nicotine's (a) anorectic
effects (Caggiula et al., 1989, 1991), (b) corticosteroneelevating effects (Caggiula et al., 1991), (c) tachycardiac
effects (Epstein et al., 1991), and (d) analgesic effects
(Caggiula et al., 1993; Epstein et al., 1989). These researchers found evidence of nicotine tolerance in subjects tested in
the same context in which they had received repeated
administrations of nicotine. However, these subjects failed
to display tolerance effects when they were tested in the
presence of a novel set of contextual stimuli (challenge test).
Thus, the researchers concluded that these subjects had
developed a context-dependent, or associative, form of
nicotine tolerance.
Alternatively, when animals are tested in a novel environment, an apparent loss of tolerance also may be accounted
for by the presence of novelty-induced stress in the test
situation. That is, environmental stress may interact with a
drug dose to augment the drug effects being measured. For
example, exposure to a novel environment can elevate
corticosterone levels (e.g., Caggiula et al., 1993) or induce
analgesia (e.g., Netto, Siegfried, & Izquierdo, 1987; Sher-

Antonio Cepeda-Benito, Jose Reynoso, and Eric H. McDaniel,

Department of Psychology, Texas A&M University.
This research was supported by Grant DA10576-01 from the
National Institute on Drug Abuse. We thank Anissa Norrell,
Thomas Cross, Andrea Szabo, Michael Drake, Eddy Badrina, Paige
Bonner, Amber Shipp, Enrique Pizana, Sarah Manning, and Jason
Murphy for help in conducting Ihe experiment.
Correspondence concerning this article should be addressed to
Antonio Cepeda-Benito, Department of Psychology, Texas A&M
University, College Station, Texas 77843-4235. Electronic mail
may be sent to acb@tamu.edu.
248

TOLERANCE TO NICOTINE ANALGESIA IN THE RAT

man, 1979). Various investigations have demonstrated that

some examples of context-specific drug tolerance may
reflect the presence of stress effects rather than the presence
of absence of associative tolerance effects (e.g., Bardo &
Hughes, 1979; Sherman, 1979; Sherman, Proctor, & Strub,
1982).
In the two studies that investigated the development of
tolerance to the analgesic effects of nicotine (Caggiula et al.,
1993; Epstein et al., 1989), the subjects were allowed to
practice the test response during the tolerance development
phase. This method could be problematic because animals
permitted to practice a task while drugged can develop more
tolerance to the disruptive effects of the drug on performance than animals given as much drug but without the
opportunity for drugged practice (sec reviews by Goudie &
Demellweek, 1986; Wolgin, 1989; Young & Goudie, 1994).
Some investigators have argued that this intoxicatedpractice effect represents the operation of instrumental
conditioning resulting in the learning of specific compensations for drug-induced disruptions in performance, that is,
behavioral contingent tolerance (e.g., LeBlanc, Gibbins, &
Kalant, 1973). In all fairness, some theorists do not conceptually differentiate between instrumental and associative
forms of drug tolerance (see Poulos & Cappell, 1991).
Our main objective was to obtain contextual nicotine
tolerance effects by using procedures that minimize the
influence of novelty and instrumental confounding factors
during testing. Given that nicotine has analgesic properties
(e.g., Rogers & Iwamoto, 1993), we modeled our procedures
after those of studies that have produced robust associative
tolerance to morphine's analgesic effects (e.g., Carter &
Tiffany, 1996; Cepeda-Benito & Tiffany, 1992, 1996a,
1996b). In these associative tolerance studies, the influence
of novelty-induced stress was controlled or minimijed by
extensively prchabituating the animals to the experimental
and tolerance assessment procedures (sec also Ramsay &
Woods, 1997). These associative tolerance investigations
also showed that intoxicated practice is not necessary for the
production of robust associative tolerance (e.g., CepedaBenito & Tiffany, 1992).

Study 1
We designed Study 1 to verify that we could assess
nicotine's antinociceptive effects in a dose-dependent manner with tail-flick and hot-plate devices. Although it is
generally accepted that nicotine reliably produces analgesic
effects in rats, in some cases the antinociceptive effects of
nicotine in the tail-flick test have not been observed (e.g.,
Yang, Wu, & Zbuzek, 1992). Unlike previous dose-related
analyses (Caggiula, Epstein, Perkins, & Saylor, 1995; Rogers & Iwamoto, 1993), our analyses attempted to minimize
stress factors during the test session by extensively prehabituating the rats to the experimental procedures and the test
environment.
Epstein et al. (1989) demonstrated the presence of tolerance of nicotine's analgesic effects by snowing that with
repealed administrations, nicotine lost its ability to increase
tail-withdrawal latencies. In order to establish the specificity

249

or generality of this reported nicotine tolerance effect in

tail-flick reflexes, we measured responses mediated both
spinally (tail flick) and supraspinally (hot plate).

Method
Subjects.
The subjects, 32 experimentally naive, male SpragueDawley rats, weighed between 300 and 350 g at the beginning of
the experiment. They were housed individually in plastic cages
with a bed of wood shavings. The colony was constantly illuminated, and the rats had continuous access to food and water in their
home cages.
Drugs. Nicotine bitartrate was dissolved in physiological saline to produce the following nicotine concentrations: 0.125, 0.25,
0.375, 0.50, 0.60, and 0.75, mg/ml. All solutions were subcutaneously injected in the scruff of the neck (1 ml/kg of body weight).
Tail-jllck analgesia assessment. The tail-nick test measures the
latency for the rat to move its tail away from a hot beam of light
(e.g., Cepeda-Benito & Tiffany, 1996a). A rat was restrained in an
opaque cylinder (6.8 x 22 cm) that had a Plexiglas base (5.5 X 22
cm). Ventilation holes were made on top of and in front of the tube.
The rat's tail protruded from the back of the tube and was placed in
a grooved plate such that the tail was directly under the light source
(HTC tail flick, model 33B). When the rat moved its tail away from
the light beam, a photosensitive cell tripped a timer and the
tail-flick latency was automatically recorded. To avoid interactions
between tail area stimulated and degree of analgesia (Yoburn,
Morales, Kelly, & Inturrisi, 1984), each assessment was the mean
of three consecutive trials with the location of the beam being
varied among the proximal, middle, and distal thirds of the rat's
tail. The beam intensity was adjusted such that undrugged animals
nicked their tails at about 4 s. A 15-s limit was used to prevent
damage to the tail. The tester was blind to the rat's group condition.
Hot-plate analgesia assessment. The hot-plate test measures a
rat's latency to lick a paw or jump (e.g., Caggiula el al., 1995;
Cepeda-Benito & Tiffany, 1996b; Krank, 1987). A rat was confined
to the hot-plate's surface in a chamber (30 X 30 X30 cm) with a
clear Plexiglass lid. The hot plate consisted of a metal surface
thermostatically controlled to a constant temperature of 50 C
(IITT hot plate, model 35D). Two independent observers, blind to
the rat's group condition, timed to the nearest hundredth of a
second each rat's latency to either lick a paw or jump, whichever
came first (e.g., Krank). The response latency was the mean of the
two observations. The median difference between observers was 1.47 s.
Rats were removed from the hot plate as soon as they either licked a paw
or jumped. Animals that neither licked a paw nor jumped after 60 s were
removed from the apparatus to prevent tissue damage.
Tail-flick prehahituation and testing phase. After 1 week of
accommodation to their new home environment, the rats were
weighed once daily for 3 days and weighed twice daily for an
additional 3 days. Rats were then habituated to injection and
mock-testing procedures. Rats received five saline injections paired
with a distinctive context and five saline injections paired with their
home cage environment. The interval between distinctive context
injections was 48 hr. Each home cage injection was given 24 hr
after each distinctive context exposure. For distinctive context
exposures, each rat was weighed, individually carried in a small
plastic container to the distinctive context room, injected with
saline, placed inside a tube, placed in a dark cabinet, and mock
tested in the tail-flick device 4, 8, and 13 min after the injection. In
the distinctive context room, the room's light was dimmed,
apple-cinnamon air fresheners scented the holding cabinet, and
white noise was continuously played. Rats were not removed from
their holding tubes. Each mock tail-flick test consisted of placing

CEPEDA BENITO, REYNOSO, AND McDANIEL

250

the rat in the tail-flick apparatus and going through the motions of
conducting three tail-flick tests (i.e., without aiming the lighl beam
at the tail). After each mock test, the rats were returned to the
scented cabinets. Rats were returned to their home cage environments 33 min after the last mock lesl. For home cage injections, the
rats were individually weighed and placed back in their home
cages. Each rat was then removed from its home cage, injected with
saline, and returned to its cage.
Rats were tested in the distinctive context 48 hr after the fifth
distinctive context exposure session (or 24 hr after the last home
cage injection). Rats were randomly divided into five groups, with
one group receiving saline and each remaining group receiving a
different test dose of nicotine (0.25, 0.5, 0.6, and 0.75 mg/kg). Rats
were tested in actual tail-flick trials 4, 8, and 13 min after being
injected with nicotine.
Hot-plale prehabituation and testing phase. Rats began five
additional distinctive context exposure sessions 48 hr after they
were tested with the tail-flick device. Except for mock tests, these
exposure sessions were identical to the tail-flick exposure sessions.
For each mock test, the rat was removed from its tube and placed on
a nonfunctional hot plate for 60 s. The rat was then placed back
inside its tube and returned to the scented cabinet. Rats received
saline injections in their home cage environment 24 hr after each
distinctive context exposure.
Rats were tested in the distinctive context 48 hr after the last
distinctive context exposure session (or 24 hr after the last home
cage injection). Rats were randomly divided into five groups, with
one group receiving saline and each remaining group receiving a
different dose of nicotine (0.125, 0.25, 0.375, and 0.50 mg/kg).
Rats were tested in actual hot-plate trials 4, 8, and 13 min after
being injected with nicotine.

Results
Animals responded to nicotine in a dose-related manner.
Thai is, tail-flick and hot-plate latencies increased as nicotine doses were raised. Figure 1A (tail-flick test) and Figure
2A (hot plate test) depict the mean response latencies for
each nicotine dose at each testing point. Figures 1B and 2B
depict mean dose-response curves across the three testing
points. Within each testing method, mean latencies across
the assessments conducted 4, 8, and 13 min after the nicotine
injections were subjected to simple regression analysis
(Cohen & Cohen, 1975). Latencies were regressed on log
dose level of nicotine. These analyses showed statistically
significant effects with both the tail-flick test, tf2 = .23,
F(\, 24) = 1.12,p < .05, and hot-plate test, R1 = .50, F(\,
24) = 22.08,p< .0001.

Study 2
Study 2 was designed to produce associative tolerance to
nicotine's analgesic effects not confounded by stress or
instrumental factors. We predicted that animals receiving
nicotine explicitly paired with a distinctive context would
become more tolerant to the antinociceptive effects of
nicotine than animals for which the nicotine administrations
and context exposure sessions were explicitly unpaired. On
the basis of associative morphine tolerance investigations
(e.g., Tiffany, Drobes, & Cepeda-Benito, 1992), we also
predicted that animals receiving nicotine in their home cages

Tail-Flick Test

15
12
.50 nic
.25 nic

saline

13

8
Testing Time (min)

Dose-Response Curve

15

I
S 6

0.25

0.5 0.6 0.75

Logarithmic Scale of Nicotine Dose (mg/kg)

figure I. Tail-flick latencies. Each point represents the mean

tail-flick latency obtained for each nicotine (nic) dose at each
testing time after nicotine administration (A) and across testing
times (B). The horizontal bar represents the mean latency for
animals injected with saline. Error bars depict 5' above and below
Ihe mean.

would develop some degree of nicotine tolerance in comparison to control animals given saline.
Method
Subjects. The subjects were 35 experimentally naive, male
Sprague-Dawley rats that weighed between 225 and 275 g at the
beginning of the experiment. They were housed and maintained as
in Study 1.
Drugs. Nicotine bitartrate was dissolved in physiological saline at a concentration of 1.00 mg of nicotine per ml. All solutions
were subcutaneously injected in the scruff of the neck (1 ml/kg of
body weight). That is, like Epstein et al. (1989), we used a 1-mg/kg
nicotine dose for tolerance development and testing with the
tail-flick and hot-plate devices.
Prel-tahituation phase. After 1 week of accommodation to their
colony room, the rals were weighed once daily for 3 days, weighed
twice daily for an additional 3 days, and then weighed and injected
with saline twice daily for 8 days. All of these procedures took
place in the colony room.
Tolerance development in the tail-flick phase. The distinctive
context and procedures for distinctive context exposure sessions
were identical to those used during the tail-flick prehabituation
phase of Study 1. Animals were divided randomly into three
groups: distinctive context (DC), home cage (HC), and saline
control (SC). Each rat was given 8 injections paired with the
distinctive context and 16 injections in its home cage environment.

251

TOLERANCE TO NICOTINE ANALGESIA IN THE RAT

Hot-Plate Test

A
60
50
oj
>,

40

30

.25 nic

20
.125 nic
saline

10

13

Testing Time (min)

Dose-Response Curve

60

50
o
1 40

saline for their home cage injections. SC animals received saline in

both environments. All other experimental procedures were the
same as those used in the hot-plate prehabituation phase of Study 1.
Tail-flick and hot-plate analgesia assessments. Analgesia assessments were identical to those conducted in Study 1. Each rat
received a 1-mg/kg dose of nicotine and was tested 4, 8, and 13 min
after the injection. Tail-flick and hot-plate test sessions were
conducted 72 hr after the eighth and last nicotine context pairings,
respectively.
Data analysis. Within each testing method, the data were
studied with a one-between, one-within repeated measures analyses of variance. Mean response latency was the dependent variable,
group Condition was the between factor, and testing time was the
within factor. To control for heterogeneity of covariance across
repeated measures, we used the Greenhouse-Geisser (1959) technique to adjust the degrees of freedom for the error terms.
Following a priori directional hypothesis, we compared mean
response latencies between SC and HC rats and between HC and
DC rats.
Results

30
CD

ra 20

10

0.125

0.25

0.375 0.5

Logarithmic Scale of Nicotine Dose (mg/kg)

Figure 2. Hot-plate latencies. Each point represents the mean

hot-plate latency obtained for each nicotine (nic) dose at each
testing time after nicotine administration (A) and across testing
times (B). The horizontal bar represents the mean latency for
animals injected with saline. Brror bars depict SE above and below
the mean.

The interval between context exposures was 72 hr. During the 2

days that followed each context exposure session, rats received a
daily injection in their home cage environment. DC rats received
nicotine in the distinctive context and saline in the home cage
environment. HC rats received saline in the distinctive context and
nicotine and saline for the first and second home cage injections,
respectively. SC rats received saline in both environments.
Tolerance development in the hot-plate phase. Following the
tail-flick test sessions, animals received two daily saline injections
in their home cages for 2 days. Then, each rat was given 5
injections paired with the distinctive context and 20 injections in its
home cage environment. That is, the ratio of saline to nicotine
injections was greater in the hot-plate phase than in the tail-flick
phase. Previous research has shown that these cues can support
associative tolerance effects in HC animals (e.g., Cepeda-Benito &
Tiffany, 1995). After observing high levels of tolerance in HC rats
in the tail-flick assay, we were concerned that HC rats were using
injection cues to support associative tolerance effects. Therefore.
we reduced the likelihood of rats receiving nicotine in the presence
of injection cues.
The interval between context exposures continued to be 72 hr,
but home cage injections were administered twice daily after each
distinctive context exposure. The first of the home cage injections
consisted of nicotine for the HC animals. All other home cage
injections consisted of saline for these animals. The interinjection
interval for each daily pair of home cage injections was 4 hr. DC
animals continued to receive nicotine in the distinctive context and

Tail flick. Figure 3 depicts mean tail-flick latencies for

each group of rats at each of the three testing times. The
tail-flick latencies differed between groups and declined
with the passage of time, and this decline was not uniform
across groups. This description of the results was corroborated by a significant condition effect, F(2, 32) 16.00, p <
.001, a significant time effect, F(2, 64) = 49.99; p < .0001,
and a significant Condition X Time interaction, F(4, 64) =
4.64, p < .05. It was clear that the pairing of the distinctive
test context with nicotine produced strong context-specific
tolerance; DC rats had significantly lower latencies than HC
rats, f(21) = 3.08, p < .005. Tn turn, repeated administrations of nicotine explicitly unpaired with the distinctive
context also produced strong tolerance of the 1-mg/kg
nicotine test dose; HC rats displayed significantly lower
Latencies than SC rats, t(2l) = 5.64, p < .0001. The nature
of the interaction effect was studied by examining the
parallelism between the group profiles (Stevens, 1992). Two
post hoc, Bonferroni-adjusted, two-tailed f tests (p < .025)
confirmed that the tail-flick latency decrements for DC rats
(M = 9.02, SD = 4.46) and HC rats (M = 7.49, SD = 3.58)
were not significantly different from each other, (21) =

15

13
Testing Time (min)

Figure 3. Tail-flick context effect. Each point represents the mean

tail-flick latency for each group condition at each testing time.
Error bars depict SE above and below the mean. SC = saline
control; HC = home cage; DC = distinctive context.

252

CEPEDA-BENITO, REYNOSO, AND McDANIEL

0.95. p > .05. However, tail-flick latency decrements were

significantly higher for HC rats than for SC rats (M ~ 3.25,
SD = 3.47), f(21) = 2.88, p < .01.
Hot plate. Figure 4 depicts mean hot-plate latencies for
each group of rats at each of the three testing times. The
hot-plate latencies differed between groups and declined
similarly across groups with the passage of lime. This
description of the results was corroborated by a significant
condition effect, F(2, 32) = 12.18, p < .001, a significant
time effect, F(2, 64) = 9.25, p < .001, and no significant
Condition X Time interaction, F< 1. As in the tail-flick test,
it was clear that the pairing of the distinctive test context
with nicotine produced strong context-specific tolerance;
DC rats responded faster than HC rats, r(21) = 3.52, p <
.005. In turn, repeated administrations of nicotine in the
home cage also produced tolerance of nicotine; HC rats
displayed significantly lower latencies than SC rats, r(21) =
4.73, p< .001.

General Discussion
Study 1 replicated the finding that nicotine can produce
analgesic effects in a dose-related manner (Caggiula et al.,
1995; Rogers & Iwamoto, 1993). This effect was found with
both the tail-flick and the hot-plate devices and was indicated by log dose being a significant predictor of tail-flick
and paw-lick latencies. These results were obtained with
procedures that minimized the presence of novelty-induced
stress during testing (see also Ramsay & Woods, 1997; cf.
Caggiula et al., 1995; Rogers & Iwamolo, 1993). That is, rats
were extensively prehabituated to the testing procedures.
The data from Study 2 showed that animals that were
injected with nicotine during the tolerance development
phase were less responsive to nicotine's analgesic effects
than animals that received nicotine only during testing. This
effect was indicated by the lower tail-flick and paw-lick
latencies observed in HC and DC rats than in SC rats. The
data also revealed that animals receiving nicotine administrations explicitly paired with a distinctive environment (DC
rats) were more tolerant than animals receiving as many
nicotine administrations but explicitly unpaired with a
distinctive context (HC rats). The results appear congruent
with the hypothesis that a distinctive context may function
as a CS capable of eliciting associative tolerance effects in
anticipation of nicotine's antinociceptive effects (e.g., Siegel, 1975).
The results of these studies appear to be reliable and
nonspecific, as they were obtained across two functionally
different responses, the spinal tail-flick reflex and the
supraspinal paw-lick response. It is unlikely that the context
effect was caused by the presence of novelty-induced stress
during the tolerance assessment session because all subjects
were extensively familiarized with the experimental and
testing procedures (cf. Caggiula et al., 1989, 1991, 1993;
Epstein et al., 1989; 1991). That is, prehabituation to the
experimental and testing procedures is often regarded as an
effective process for minimizing confounding stress effects
in drug tolerance experiments (e.g., Ramsay & Woods,
1997).

60

tr 50
&
40

I
-3 30
20

13
Testing Time (min)

Figure 4. Hot-plate context effect. Each point represents the

mean hot-plate latency for each group condition at each testing
time. Error bars depict SE above and below the mean. SC = saline
control; HC = home cage; DC = distinctive context.

Our finding that a distinctive context functioned as a CS

for the development of associative drug tolerance replicates
the results of the many studies that have investigated the
development of tolerance to morphine's analgesic effects
(e.g., Cepeda-Benito & Tiffany, 1992; Tiffany & MaudeGriffin, 1988). Therefore, our results are important theoretically because they help support the generalizability of
associative morphine tolerance phenomena to other drugs.
This assertion is congruent with the statement by Poulos and
Cappell (1991, p. 402) "to go beyond morphine" when
assessing the adequacy of general models of drug tolerance.
That is, these authors defended the idea that a comprehensive theory of drug tolerance should establish a generality of
tolerance findings across different drugs (Poulos & Cappell).
We emphasize that instrumental learning could not have
played a role in the context effect because the subjects did
not practice the test response during the tolerance development phase (cf. Caggiula et al., 1993; Epstein et al., 1989).
However, this observation merits attention only if instrumental and associative drug tolerances are defined as being
conceptually different (e.g., Cepeda-Benito & Tiffany, 1995).
Conversely, Poulos and Cappell (1991) defend the idea that
all tolerance that develops to specific drug effects is,
explicitly or implicitly, behaviorally contingent. These authors theorize that, regardless of the experimenter's manipulation, drug tolerance will develop only if drug effects place
behavioral demands on specific physiological systems. That
is, in the absence of behavioral or functional disturbances,
organisms would not have a need to initiate homeostatic
restoration processes, or tolerance.
Nevertheless, from a merely methodological viewpoint,
our results at least corroborate the idea that contextual
tolerance can be obtained without assessment of a drug's
effects on behavioral responding after each drug administration during the tolerance development phase (see also
Cepeda-Benito & Tiffany, 1992; Tiffany et al., 1992).
Therefore, by systematically manipulating various parameters within the present design, future research on nicotine
tolerance could further examine the conditions under which
associative nicotine tolerance develops (see Baker & Tiffany, 1985; Poulos & Cappell, 1991).
There are two potential problems with our experimental

TOLERANCE TO NICOTINE ANALGESIA IN THE RAT

design. First, testing of the same rats first in the tail-flick test
and then in the hot-plate test did not allow us to compare
tolerance effect sizes across both testing methods. That is,
the tail-flick test was given after 8 conditioning sessions,
whereas the hot-plate test was given after 14 nicotine
context pairings. Therefore, any effect size differences
between the two methods could not be interpreted. Moreover, we did not control for the possibility that testing of the
rats in the tail-flick test first could have influenced the
hot-plate test results. These problems could have been
avoided by counterbalancing the order of testing across
measurements or by testing independent groups of rats in
each testing device. Nevertheless, we believe that it is safe to
affirm that the order of testing docs not compromise our
interpretation of the results. That is, compared to DC rats,
HC rats had a lower probability of receiving nicotine inside
the distinctive context and a higher probability of receiving
nicotine outside the distinctive context in both test sessions.
Both sets of probabilities are congruent with the prediction
that DC animals would develop a stronger context-drug
association than HC animals (see Rescorla & Wagner,
1972).
The experimental design also could have been improved
by including a placebo test control group for each of the
three conditions, SC, HC, and DC. Given that we repeatedly
paired the testing environment with nicotine for DC but not
SC and HC rats, undrugged control latencies could have
helped to clarify whether the context effects were attributable to differential changes in distinctive context baseline
latencies across groups. For example, the development of
contextual tolerance could have been mediated by a conditioned hyperalgesic response (e.g., Siegel, 1975).

References
Alexander, B. K., & Hadaway, P. K (1982). Opiate addiction: The
case for an adaptive orientation. Psychological Bulletin, 92,
367-381.
American Psychiatric Association. (1994). Diagnostic and statistical manual of mental disorders (4th ed.). Washington, DC:
Author.
Baker, T. B., & Tiffany, S. T. (1985). Morphine tolerance as
habituation. Psychological Review, 92, 78-108.
Bardo, M T., & Hughes, R. W. (1979). Exposure to a nonfunctional
hotplate as a factor in the assessment of morphine-induced
analgesia and analgesic tolerance in rats. Pharmacology Biochemistry and Behavior, 70,481^185.
Caggiula, A. R., Epstein, L. H., Antelman, S. M., Saylor, S., Knopf,
S., Perkins, K. A., & Stiller, R. L. (1993). Acute stress or
corticosterone administration reduces responsiveness to nicotine: Implications for a mechanism of conditioned tolerance.
Psychopharmacology, I I I , 499-507.
Caggiula, A. R., Epstein, L. H., Antelman, S. M., Seymour, M.,
Saylor, S., Perkins, K. A., Knopf, S., & Stiller, R. L. (1991).
Conditioned tolerance to the anorectic and corticosteroneclevating effects of nicotine. Pharmacology Biochemistry and
Behavior, 40, 53-59.
Caggiula, A. R., Epstein, L. H., Perkins, K. A., & Saylor, S. (1995).
Different methods of assessing nicotine-induced antinociception
may engage different neural mechanisms. Psychopharmacology,
122, 301-306.

253

Caggiula, A. R., Epstein, L. H., & Stiller, R. L. (1989). Changing

environmental cues reduces tolerance to nicotine-induced anorexia. Psychopharmacology, 99, 389-392.
Carter, B. L., & Tiffany. S. T. (1996). Cross-tolerance of associative
and nonassociative morphine tolerance in the rat with mu- and
kappa-specific opioids. Psychopharmacology, 123, 289-296.
Cepeda-flenito. A., & Short, P. (1997). Morphine's interoceptive
stimuli as cues for the development of associative morphine
tolerance in the rat. Psychobiology, 25, 236-240.
Cepeda-flenito. A., & Tiffany, S, T. (1992). Effect of number of
conditioning trials on the development of associative tolerance
to morphine. Psychopharmacology, 109, 172-176.
Cepeda-Benito, A., & Tiffany, S. T. (1995). Role of drugadministration cues in the associative control or morphine
tolerance in the rat. Psychopharmacology, 122, 312-316.
Cepeda-Benito, A., & Tiffany, S. T. (1996a). The failure of
noncontingent morphine exposure to disrupt context-specific
morphine tolerance. Pharmacology Biochemistry and Behavior,
54, 575-580.
Cepeda-Benito, A., & Tiffany. S. T. (1996b). Test-specific manifestations of associative tolerance to the analgesic effects of
morphine in the rat. Psychobiology, 24, 327-332.
Cohen, J.. & Cohen, P. (1975). Applied multiple regression/
correlational analysis for the behavioral sciences. Hillsdale, NJ:
Erlbaum.
Epstein, L. H.. Caggiula, A. R., Perkins, K. A., McKenzic, S. J., &
Smith, J. A. (1991). Conditioned tolerance to the heart rate of
smoking. Pharmacology Biochemistry and Behavior, 39, 15-19.
Epstein, L. H., Caggiula, A. R., & Stiller, R. L. (1989). Environmentspecific tolerance to nicotine. Psychopharmacology, 97, 235237.
Goudie, A. J., & Demellweek, C. (1986). Conditioning factors in
drug tolerance. In S. R. Goldberg & I. P. Stolerman (Eds.),
Behavioral analysis of drug dependence (pp. 225-285). New
York: Academic Press.
Greenhouse, S. W., & Geisser, S. (1959). On methods in the
analysis of profile data. Psychometrika, 24, 95112.
Grisel, J. E., Wiertelak, E. R.'Watkins, L. R., & Maicr, S. F. (1994),
Route of morphine administration modulates conditioned analgesic tolerance and hyperalgesia. Pharmacology Biochemistry and
Behavior, 49, 1029-1035.
Krank, M. D. (1987). Conditioned hyperalgesia depends on the
pain sensitivity measure. Behavioral Neuroscience, 101, 854
857.
LeBlanc, A. E.. Gihhins, R. J., & Kalant, H. (1973). Behavioral
augmentation of tolerance to ethanol in the rat. Psychopharmacologia. JO, 117-122.
Netto, C. A., Siegfried, B., & Izquierdo, I. (1987). Analgesia
induced by exposure to a novel environment in rats: Effect of
concurrent and post-training stressful stimulation. Behavioral
and Neural Biology, 48, 304-309.
Pomerleau, O. F. (1995). Individual differences in sensitivity to
nicotine: Implications for genetic research on nicotine dependence. Behavior Genetics, 25, 161-177.
Poulos, C. X., & Cappell, H. (1991). Homeostatic theory of drug
tolerance: A general model of physiological adaptation. Psychological Review, 98, 390-408.
Ramsay, D. S., & Woods, S. C. (1997). Biological consequences of
drug administration: Implications for acute and chronic tolerance. Psychological Review, 104, 170-193.
Rescorla, R. A., & Wagner, A. R. (1972). A theory of Pavlovian
conditioning: Variations in the effectiveness of reinforcement
and nonreinforcement. In A. H. Black & W. F. Prokassy (Eds.),
Classical conditioning 11: Current research and theory (pp.
64-99). New York: Appleton-Century-Crofts.

254

CEPEDA-BENITO, REYNOSO, AND McDANIEL

Rogers, D. T., & Iwamoto, E. T. (1993). Multiple spinal mediators

in parenteral nicotine-induced antinociception. Journal of Pharmacology and Experimental Tlicrapeutics, 267, 341-349.
Sherman, J. E. (1979). The effects of conditioning and novelty on
the rat's analgesic and pyretic responses to morphine. Learning
and Motivation, 10, 381-418.
Sherman, J. E., Proctor, C., & Strub, H. (1982). Prior hotplate
exposure enhances morphine analgesia in tolerant and drugnaive rats. Pharmacology Biochemistry and Reluivior, 17, 229232.
Siegel, S. (1975). Evidence from rats that morphine tolerance is a
learned response. Journal of Comparative and Physiological
Psychology, 89, 498-506.
Stevens, i. (1992). Applied multivariate statistics for the social
sciences (2nd ed.). Hillsdale, NJ: Erlbaum.
Tiffany, S. T., Drobes, D. J., & Cepeda-Benito, A. (1992).
Contribution of associative and nonassocialive processes to the
development of morphine tolerance. Psvchopharmacology, 109,
185-190.
Tiffany, S. T., & Maude-Griffin, P. M. (1988). Tolerance to
morphine in the rat: Associative and nonassociative effects.
Behavioral Neuroscience, 102, 534-543.
Trujillo, K. A., & Akil, H. (1995). Excitatory ainino acids and drugs
of abuse: A role for W-methyl-D-asparate receptors in drug

tolerance, sensitization and physical dependence. Drug and

Alcohol Dependence, 38, 139-154.
Wolgin, D. L. (1989). The role of instrumental learning in
behavioral tolerance to drugs. In A. J. Goudie and M. W.
Emmit-Ogiesby (Eds.), Pxvchoactive drugs: Tolerance and
sensitization (pp. 17-114). Clifton, NJ: Humana Press.
Yang, C., Wu, W., & Zbuzek, V. K. (1992). Antinociceptive effect
of chronic nicotine and nociceptive effect of its withdrawal
measured by hot-plate and tail-flick in rats. Psvchopharmacology, 106, 417-420.
Yobiirn, B. C., Morales, R.. Kelly, D. D., & mturrisi, E. C. (1984).
Constraints on the tail-flick assay: Morphine analgesia and
tolerance are dependent upon locus of tail stimulation. Life
Sciences, 34, 1755-1762.
Young, A. M., & Goudie, A. J. (1994). Adaptive processes
regulating tolerance to behavioral effects of drugs. In F. E.
Bloom & D. J. Kupfer (Eds.), Pxychopharmacology: The fourth
generation of progress (pp. 657-811). New York: Raven Press.

Received November 18, 1997

Revision received March 24, 1998
Accepted March 25, 1998

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