Psychological Reporfs, 1996,: in Al.
Psychological Reporfs, 1996,: in Al.
Psychological Reporfs, 1996,: in Al.
B. F. PETRIE
New Mexico Tech
Method
Subjects.-There were 10 male and 10 female Wistar rats of Charles
River Canada, Inc. origin, in both the isolated and colony groups. The ani-
mals were raised from weaning (21 days of age) in their respective environ-
ments, and were 113 days of age when the experiment started.
Apparatus.-Isolated rats were housed from weaning in standard rat
cnges (18 x 25 x 18 cm) with sheet metal walls that prevented visual contact
w ~ t hadjacent animals. To collect wastes, paper was placed on trays under
the cages. These rats received fluids through stadess steel drinlung tubes
from plastic bottles (Girton, Mdville, PA) fastened on the outside of each
cage. Purina rat chow was provided ad libitum by means of inside feeders
(11 x 13 x 5.5 cm). Bottles attached to two empty control cages allowed for
the calculation of spdlage and evaporation that might occur in a 24-hr. pe-
riod.
Colony rats lived together from weaning in an open-topped wooden
box with a floor area of 8.8 m2. The box contained a layer of kiln-dried ce-
dar shavings (Hyon Bedding) and two large open-topped metal cages (40 x
25 x 18 cm) from which two feeders (24 x 12.5 x 5 cm) containing Purina
rat chow were hung. The animals had continuous access to a common drink-
ing source. To drink, each animal climbed a pole (41 cm long; 4 cm circum-
ference) triggering a videotaped recording of that rat's identhing hair dye
mark (L'Oreal Excellence-Napoli Black, Cosmair Canada, Inc.) and drank
from one of two nipples (Edstrom Industries, Lnc. A115 Adjustable Flow
WISTAR RATS' ORAL MORPHINE CONSUMPTION 393
Valve No. 10443). The computer-controlled system noted the weight of each
of two fluids consumed (resolution of 0.1 gram) for each visit to the site by
a rat. The time and duration of the visit were also recorded. Rats learned to
operate the system within three days of its introduction to the colony. This
drinking system is described fully elsewhere (Petrie, et al., 1985).
The white fluorescent lighting in both environments was on a 12-hr.
light-dark cycle controlled by a single timer (Tork Time Switch Model
7102). Red lights (Sylvania 25- and 60-watt bulbs) were on in both environ-
ments at all times.
Procedure.-The animals were placed in individual cages or the colony
at 21 days of age. At 86 days of age the colony rats were dye-marked for
identification and at 99 days of age the control animals had a second bottle
of fluid attached to their cages. Lntake testing began at 113 days of age, and
all rats were weighed at 114 days of age. Intake testing concluded when the
animals were 142 days of age and all rats were weighed again and kdled at
143 days of age.
During intake testing all animals were given 24-hour access to tapwater
and the alternative experimental fluid. The seven phases of testing were each
four days in length, and unhke experiments with the older apparatus, no
data were lost due to malfunctioning equipment.
In the first and seventh phases, access to tapwater and to a 10% su-
crose solution was provided to assess whether housing conditions had any
effect on consumption of sucrose.
The second phase compared the intake of tapwater to that of a solution
of 10% sucrose with 0.05 mg/ml quinine sulfate, to check for the effects of
housing on preference for bitter-sweet solutions. To the human palate, this
sucrose-quinine solution tasted the same as the sucrose-morphine solution
used in the 0.25 morphine hydrochloride phase.
Phases 3 through 6 entailed continuous access to water and to progres-
sively decreasing concentrations of morphine hydrochloride (MHCI) in 10%
sucrose. Phase 3 included 1.0 mg MHCVml 10% sucrose. Phase 4 consisted
of 0.5 mg MHCl/ml 10% sucrose. Phase 5 involved 0.25 mg MHCl/ml 10%
sucrose. Phase 6 comprised 0.125 mg MHCVml 10% sucrose (see Table 1).
Left-right positions of water and the experimental fluid were reversed
every two days in both environments.
Results
Two-way repeated-measures analyses of variance corrected for multiple
comparisons were carried out separatelv for each phase on the proportion of
experimental fluid to total fluid consumed; on mdhgrams of experimental
substance ingested per kilogram of body weight; on grams of experimental
fluid consumed; and on total fluid consumption (grams). Significant interac-
TABLE 1
IN EACHPHASETHE RATSO F BOTHGROUPS
ORDEROF EXPERIMENTALPHASES:
HADA CHOKEBETWEEN TAPWATERA N D THE SOLUTION
DESCRIBED
p < .05). Colony males in Phase 3 drank significantly more total fluid than
&d isolated females (df=34, p<.05). In Phase 4, colony females consumed
significantly more total fluid than did isolated females (df =33, p < .05), iso-
lated males (df=33, p < ,051, and colony males (df=33, p < .05). Colony
males in Phase 4 consumed significantly more total fluid than did isolated
females (df=33, p < .05). In Phase 7, isolated males drank significantly more
total fluid than d d colony males (df = 33, p < .05). Colony females drank sig-
nificantly more total fluid than did colony males (df= 33, p < .05); see Table
2.
TABLE 2
AVERAGE
NUMBER
OF G W S OF FLUIDCONSUMED DNLY:FIRSTSUCROSE-MORPHINE
STUDY
Dzscusszon
The results of this study do not replicate some of the earlier research
on housing and morphine consumption. Females drank significantly more of
the experimental fluids (mg/kg) in all of the four sucrose-morphine phases.
This is consistent with Hadaway, et al.'s (1979) observation that females gen-
erally drank more morphine solution than dId males.
The major finding of the earlier research on housing that rats housed in
the colony at the time of testing drank significantly less morphine than did
the isolated rats was not confirmed. In fact, during- the 1.0-rng M H C V d
10%-sucrose phase, the colony rats in the present study drank signi€icantly
more than did the isolated animals on all four measures, although the mag-
nitude of the differences was small. There were no significant differences in
morphine consumption between the colony and caged animals in either the
0.5-mg, 0.25-mg, or 0.125-mg MHCVml 10%-sucrose phases.
The possibility exists that either the results of this study or the three
studies conducted with the older technology could be artifacts of the mea-
surement procedure. Therefore, the second study reported was designed
without any kind of automated equipment to measure the fluid intake of the
colony animals. Although this way of measuring fluid intake precluded the
gathering of any individual fluid-consumption patterns in the colony rats,
the amount of fluid taken from colony reservoirs could be weighed and com-
pared to the amount of fluid being removed from the control cages during
the same time interval.
Method
Subjects.-There were 10 male and 10 female Wistar rats of Charles k v -
er Canada, Inc. origin, in both the isolated and colony groups. The animals
were raised from weaning (21 days of age) in their respective environments
and were 113 days of age when the experiment started.
Apparatus.-Except for the automated drinkmg system, the apparatus
used in both the isolated and colony conditions was identical to that used in
the previous study. In the colony condtion the animals had continuous ac-
cess to a common drinking source. The 41-cm pole present in the first study
was removed and the automated drinkmg system disabled. At the base of
the wall on which the drtnlung system was hung, two holes were dr~lledand
two nipples (Edstrom Industries, Inc. AL 113 Adjustable Flow Valve-No.
10441) were positioned in these holes. From each nipple ran a one metre
length of plastic tubing (Tygon R-3603) into a one-gallon plastic reservoir sit-
uated on a 60-cm high stool. Each resewoir was f i e d with the assigned
fluid and weighed daily. Beside each reservoir there was another identical
container filled with thk same fluid, with an identical length of hosing lead-
ing down to a similar nipple at the end. The nipple for this container was
placed against the outside wall of the open-topped wooden box and was not
touched by the rats. These containers served as control reservoirs and allow-
ed for the calculation of any evaporation that might occur in a 24-hr. period.
Procedure.-Except for the automated drinking system, the experimen-
tal protocol followed was identical to the procedure in the previous study.
Results
Although there could be no inferential analysis of the colony group's
data since there was no way of identifying indvidual animals' fluid con-
sumption, the averages
- obtained indicate that the colony animals outdrank
the isolated animals during the 1.0-mg sucrose-morphine phase (M=4.3 gm.,
M=0.3 gm. per day); whereas the isolated animals outdrank the colony ani-
mals during the 0.5-mg sucrose-morphine phase (M=9.4 gm., M=3.O gm.
per day); the 0.25-mg sucrose-morphine phase (M= 17.4 gm., M = 10.9 gm.
per day), and the 0.125-mg sucrose-morphine phase (M= 44.4 gm., M = 33.1
gm. per day); see Table 3.
The differences between the two groups in this study were never as
WISTAR RATS' ORAL MORPHINE CONSUMPTION 3 97
TABLE 3
NUMBER
AVERAGE OF G W S OF FLUIDCONSUMED DAILY:
SECOND
SUCROSE-MORPHINE
STUDY
large as the ddferences observed in the eight male and eight female rats that
were maintained in their original environments in the Alexander, et al.
(1981) study. The continuously isolated animals in that study drank up to
seven times as much sucrose-morphine as the rats that lived in the colony
from weaning to the end of the study.
'
The results of this study are consistent with those obtained tn the first
study using the automated system to monitor colony drinking. Therefore,
the results of the two studies reported indicate that the results of the earlier
housing research were not replicated.
(1981) study drank less sucrose, as measured in Phases 1 and 7, than did the
Wistars used in the two present studies. Clearly the isolated animals in the
two studies reported here were avoiding the consumption of morphine.
This finding leads to the issue of differences between rat strains in psy-
chopharmacological research. The possibhty exists that the Wistar rats used
in the two studies reported here ddfered from the Wistar rats used in the
Alexander, et al. (1981) study. The Alexander, et al. study that the research
reported here was designed to replicate was published in 1981. However,
the research for that publication was done from April to July in 1979. In
November 1979, the animal supplier changed outbred Wistar rat colonies.
Therefore the Wistars used in the Alexander, et al. (1981) study were Old
Colony Wistars, while the rats used in the present research were New Col-
ony Wistars.
The reasons for the changing of the colonies included health, housing,
and breeding. A number of viruses that compromised long-term studies
were eradicated; the quality of the animal housing was improved while breed-
ing ratio and productivity were increased (J. Gayer, Managing Director,
Charles River Canada, h c . , St. Constant, Ont., personal communication,
May, 1985).
It is well known that there are strain differences in responsivity to drugs
in both mice and rats (Bardo & Gunion, 1982; C o h s & Whitney, 1978;
Horowitz, Whitney, Smith, & Stephan, 1977; Oliverio, Castellano, Racagni,
Spano, Trabucchi, & Cattabeni, 1978; Shearer, Creel, & Wilson, 1973). It is
quite possible that, if a genetic alteration were inadvertently introduced when
the animal supplier changed from Old Colony to New Colony animals, this
genetic shift could be manifest in New Colony animals that responded dif-
ferently to psychoactive substances than did Old Colony animals.
Experimental evidence suggests that the Old and New Colony Wistars
respond differently to equivalent levels of psychoactive substances. Ton,
Blair, Holmes, and Amit (1983) compared the effects of chronic naltrexone
injection on amphetamine locomotor activity on indvidually housed Old
and New Colony male rats. Rats were tested for locomotor activity in the
open field with or without white noise. During the testing period the New
Colony animals showed a significant attenuation in amphetamine locomotor
activity in the absence of noise only. In contrast, chronic naltrexone signifi-
cantly decreased amphetamine activity in Old Colony animals only under
noise conditions. Ton, et al. (1983) thought that the differential effects may
reflect predi~~ositional differences across animal populations in the modula-
tion of dopamine function by opioid peptides via opiate receptors.
Another ddference between Old and New Colony Wistars includes lev-
els of voluntary alcohol ingestion, with consumption by New Colony Wistars
being greatly attenuated when compared to intake of Old Colony rats (F. J.
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