Physiology & Behavior, Vol. 24, pp. 337-340. PergamonPress and Brain Research Publ., 1980.

Printed in the U.S.A.

Estrogenic Effects on Behavioral

Thermoregulation and Body Temperature of
Rats I

C H A R L E S W. W I L K I N S O N 2, H A R R Y J. C A R L I S L E , A N D R O B E R T W. R E Y N O L D S

University o f California, Santa Barbara, CA

R e c e i v e d 24 O c t o b e r 1978

WILKINSON, C. W., H. J. CARLISLE AND R. W. REYNOLDS. Estrogenic effects on behavioral thermoregulation

and body temperature of rats. PHYSIOL. BEHAV. 24(2) 337-340, 1980.--Estradiol benzoate increases responding for
heat reinforcement of ovariectomized female and intact and castrate male rats placed in the cold. In females, both pre- and
postsession rectal temperatures are depressed by the steroid. The maintenance of low temperatures despite increased heat
intake suggests that body temperature is regulated at a lower level as a result of estradiol.induced increases in heat loss
or decreases in heat production.
Estradiol benzoate Thermoregulatory behavior Temperature Energy balance

O V A R I A N steroids, particularly estradiol, have pronounced
effects on two facets of the behavioral regulation of energy
balance in rats. Administration of estradiol to ovariec-
tomized rats results in reduced food intake and increased
locomotor activity [22]. Progesterone, in contrast, has little
independent effect, although it is potent in suppressing the
effects of estrogens [22]. The influence of estradiol on behav-
ioral thermoregulation, a third facet of energy balance con-
trol, has not been investigated directly even though it has
been known since the last century that basal body tempera-
ture fluctuates as a function of ovarian cycles [10]. Indices of
the effects of ovarian steroids on behavioral thermoregula-
tion have been obtained only by measurements of cyclic var-
iations in nestbuilding by rats and thermal comfort ratings by
women. In neither study were hormone levels manipulated
directly, and neither study allowed subjects direct, quantifi-
able control over their thermal environment.
Rats maintained at 21 ° C were reported to build smaller
nests at estrus than during diestrus [15], but nestbuilding,
particularly at this moderate temperature, cannot be as-
sumed to serve a purely thermoregulatory function. It is a
maternal response that occurs even at very high ambient
temperatures in postparturient females [15], and occurs even
if the young are permanently removed immediately after
birth [19]. It has a humoral basis in that it can be rapidly
elicited in virgin females by injections of plasma from new
mothers [ 19]. Since the maternal and thermoregulatory com-
ponents of nestbuilding may both be affected by ovarian
steroids, the measure is confounded as an index of steroidal
thermoregulatory actions. The failure of rats to build large
nests at estrus does not lead to definite conclusions about
thermoregulation for other reasons. Rats dramatically in-
crease their locomotor activity at estrus [3], and running is
incompatible with nesting. Since either huddling in a large
nest or running would raise the rat's body temperature, the
former by slowing the rate of heat loss and the latter by
increasing heat production, evidence of smaller nests during
estrus does not provide an unambiguous indication of cyclic
alterations of thermoregulatory parameters.
In the other previous study of thermoregulatory behavior,
women submerged in a constant-temperature water bath
rated the pleasantness or unpleasantness of hand contact
with a variety of thermal stimuli [8]. Cool temperatures that
were found to be unpleasant during the luteal phase of the
menstrual cycle were reported to be very pleasant during the
follicular phase. The authors interpreted their results to indi-
cate an elevation in set-point during the post-ovulatory phase
of the cycle.
It has been demonstrated in several mammalian species
that administration of progesterone results in a rise in body
temperature [9, 14, 16, 23], and estrogens have generally
been reported to elicit a decrease in temperature [4, 9, 14]. In
human menstrual cycles, intervals of elevated temperatures
are coincident with high plasma concentrations of proges-
terone, and periods of lower temperatures reflect high ratios
of estradiol to progesterone in the circulation [21]. In rats,
temperatures have been reported to be low at proestrus-
estrus relative to diestrus or pseudopregnancy [3,17], al-
though there is one report of elevated temperatures at estrus
[16]. The relationship between cyclic hormonal variations
and temperature fluctuations in the rat is obscured by the
brevity of the estrous cycle and the lag between hormonal
fluctuations and their behavioral and physiological manifes-
tations. There is an approximately 12-hr lag between the

1Supported by USPHS grants MH 11342 and HD 10473.

zPresent address and address for reprints: Department of Physiology, University of California, San Francisco, CA 94143.

C o p y r i g h t © 1980 Brain R e s e a r c h Publications Inc.--0031-9384/80/020337-04502.00/0

 ~s
morning of proestrus when estradiol and progesterone con-
centrations are maximal and minimal, respectivel3, 12, 5, 13],
and the appearance of vaginal cornification, sexual receptiv-
ity, increased running, and decreased food,intake associated
with estrogenic action, but there is no reason to assume that
all estrogenic effects will follow the same time course.
We therefore used operant techniques to determine di-
rectly the effects of estradiol on behavioral thermoregulation
of female rats. Estradiol benzoate was administered to
female rats both before and after ovariectomy, but the ques-
tion of primary interest was the effect of estradiol in castrate
females in which it is unopposed by endogenous ovarian
progesterone. Males were also tested to determine the sex-
specificity of estrogenic actions.
METHOD
Nine female and nine male adult Sprague-Dawley rats
were housed individually and maintained on ad lib food and
water at an ambient temperature of 23 + 2' C. During the
experimental sessions the rats were placed in a cage inside a
commercial freezer maintained at - 7 _+ 2 ~'C. Extending into
the cage was a Plexiglas lever, the depression of which ac-
tivated two infrared heat lamps mounted at each side of the
cage and focused in front of the lever. The total output of the
lamps was regulated at 300 W, producing a radiant flux den-
sity of 180 mW/cm e in the center of the cage. The heat lamps
remained on as long as the lever was depressed. In this way,
the rats controlled both the frequency and duration of heat
reinforcement. This method provides for efficient and pre-
cise thermoregulation [6,71. The animals were given several
practice sessions to stabilize responding before the data ses-
sions began.
Estradiol benzoate (EB) was administered via subcutane-
ous implantation of a single Silastic capsule in the scapular
region. The capsules were constructed of Silastic Medical-
Grade Tubing (Dow Corning) I.D. 1.57 mm, O.D. 2.41 ram.
Capsules had an internal length of 5 ram, were filled with
2.1-2.3 mg EB and were sealed at the ends with Silastic Type
A Medical Adhesive. In order to determine the levels of
circulating estradiol that resulted from implantation of these
capsules, concentrations of estradiol in individual plasma
samples from another group of five ovariectomized females
of comparable weight implanted with the same-sized EB
capsules were determined by radioimmunoassay. Chroma-
tography and assay procedures followed those of Abraham
et al. [!1 and were modified by L. D. Keith for measurement
of individual rat plasma samples. The values for individual
animals ranged from 90 to 165 pg/ml estradiol with a mean of
120 pg/ml. Peak plasma estradiol concentrations in rats at
proestrus have been reported to be between 50 [12] and 110
[20] pg/ml.
Capsule implantations and bilateral ovariectomies and or-
chidectomies were performed under ether anesthesia. After
each surgical manipulation, animals were allowed two to
three weeks for recovery and stabilization of hormone
levels, and then were given one additional practice session
one week before the first data session. Rats were tested in
three 3-hr sessions at weekly intervals under each condition.
Experimental sessions always occurred at the same time of
day during the dark phase of the rats' light cycle, and the rats
were shaved and food-deprived three hours prior to each
session. Seconds of heat reinforcement for each session
were recorded and averaged for each animal for each condi-
tion. Pre- and postsession rectal temperatures were obtained
WILKINSON, ('AR[ASLI--, ANi) Rtlh N()i I ~
with a Model 46 Telethermometer (Yellov, Springs in>!rtr
ment Co.) and a No. 402 probe inserted 5 cm t\~r ~0 see P:c
and postsession body weights were also recosdcd
Statistical comparisons between data l'rom EB-.treate~i
castrate animals and data from the same animals after ~ap-
sule removal were made with Student's t-test for correlated
groups. All other comparisons between intact animals and
the same rats after EB implantation or castration wcrc made
with Dunnett's t-test for multiple comparisons with a contrail
condition.
RESULTS AND DISCUSSION
The two treatment conditicns of primary interest are the
ovariectomized females with (castrate-EB implant, Fig. I A)
and without (castrate-implant out) EB capsules, in which the
effects of the hormone can be observed in the absence of
endogenous ovarian steroids. EB treatment of the ovariec-
tomized rats resulted in a significant i n o e a s e (p<0.01) in
responding for heat reinforcement. There was no difference
between pre- and postsession rectal temperatures within
either of the two groups, indicating precise behavioral ther-
moregulation. However, comparisons between the two
groups reveal that EB treatment of castrate females lowered
both pre- (,0<0.05) and postsession ¢o<0.01) rectal tempera-
tures (Fig. IA). EB treatment increased responding for heat,
yet body temperature did not increase but was maintained at
the presession low level. This finding suggests that estradiol
lowers the temperature set-point in females, and also re-
duces the animals' physiological capacity to maintain body
temperature even at that lowered set-point when faced with
cold ambient temperatures. The animals compensate for re-
duced physiological capacity by increased responding for
heat [7]. The estrogen-induced change in physiological ca-
pacity could take the form of either a reduction of metabolic
heat production or an increase in heat loss.
Neither EB treatment of intact females nor ovariectomy
alone had any consistent effect on heat intake (Fig. IA). Two
of five intact females treated with EB did exhibit significant
increases in responding, but three showed no change.
Neither ovariectomy nor EB treatment alone had any effect
on rectal temperature of females. These results suggest thai
ovarian progesterone opposes the effect of estradiol on
thermoregulatory behavior as well as on body temperature,
since ovariectomy removes the primary source of both hor-
mones. EB treatments significantly elevated heat intake in
both intact (/)<0.02) and castrate (p<0.05) males (Fig. IB),
Castration alone also increased responding q)<0.05) in
males. EB capsules elevated presession temperatures of in-
tact males (p<0.01) but had no effect on castrates. Testos-
terone has been shown to depress rectal temperature in
female rats [18] and may be responsible for the low tempera-
tures of the intact, untreated males.
Given that estrogens reduce food intake and body weight
in rats [22] and that deprivation-induced weight loss results
in increased responding for heat reinforcement [11]. it is a
plausible hypothesis that the observed EB effect on behav-
ioral thermoregulation is secondary to and dependent upon
EB-elicited reduction of body weight. If this were the case a
negative correlation should exist between presession body
weight and seconds of heat reinforcement for each animal
across all sessions. Calculation of linear correlation coeffi-
cients for individual females in this study yielded values o f t
ranging from +0.04 to +0.85, which do not support this hy-
pothesis. Because adipose tissue lost or gained as a result of
ESTROGENS AND BEHAVIORAL THERMOREGULATION 339

A ~5 the implantation or removal of EB capsules could affect insu-

Females lation and energy stores, changes in body weight between
treatment conditions were compared with changes in re-
2O
sponding. EB treatment of castrate females resulted in a
.E
,e 2.0 _+ 0.2% decrease in body weight, but a 68.4 _+ 19.3% in-
crease in responding for heat reinforcement. Removal of EB
capsules from ovariectomized rats resulted in increases in
body weight, primarily adipose tissue. Correlation of abso-
E I -] 38.5 ~ lute or percentage increases in body weight with decreases in
E
@ 0 37.5 N
responding yielded r values less than 0.1. Therefore neither
body weight nor weight change are related to responding for
-- 5
g heat reinforcement of female rats. Correlations of body
3: -~ 3T.O '~"
weight with heat intake in males, however, ranged from
r = - 0 . 0 5 to -0.89, suggesting that EB-induced increases in
Intact Ird+oct- Costrohl Costro'~- CaMrate-
EB i m p k ~ EB implant implant out responding by males may be functionally related to altera-
tions in food intake.
B 2~ It also might be argued thai estradiol-induced increases in
responding for heat are an artifact of increases in locomotor
2O activity. If this were the case, then EB-treated rats should
make proportionately more responses than untreated ani-
c
mals, and seconds of heat reinforcement per response should
j m be lower in EB-treated, castrate females than in the same
animals after capsule removal. In fact, the reverse is true
+ e
38.5 ~o (p<0.05). Mean response duration was 15.56 sec during EB
e I0
~° treatment and 13.12 sec after capsule removal. EB implanta-
"Z tion produced no significant change in the average response
g
5 duration of males.
g
"I-
3ZO "" This study demonstrates that EB treatment of ovariec-
tomized females produces marked increases in responding
Intact Intoet- C~trote CoMratl- C~trate- for heat reinforcement which are independent of EB-elicited
EB impl~d' EB imp~rd implantout
weight losses. The failure of the increased heat intake to
raise body temperature implies that EB facilitates heat loss
FIG. 1. Heat reinforcement and body temperature of (A) females or inhibits heat production in these animals. In males, the
and (B) males. Shaded bars represent seconds of heat reinforcement failure of EB to lower body temperature and the negative
(seconds of lever depression) per minute in the cold chamber for correlation between heat intake and body weight suggest that
each treatment condition. Results are expressed as the average
number of sec/min _+ SEM for all animals in a given condition. the mechanisms producing the EB-elicited increases in re-
The number of rats in each condition is indicated by the numerals in sponding are not identical to those acting in females.
the shaded bars. Treatment order was different for different animals,
and all rats did not receive all treatments. Unshaded bars represent
average pre- and postsession rectal temperatures _+ SEM for ani-
mals in each condition. Presession temperatures are represented on
the left and postsession on the right within each condition.

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