Neurobiologyof Aging, Vol. 4, pp. 187-190, 1983. ©Ankho InternationalInc. Printed in the U.S.A.

/3-Endorphin: Effect on Thermoregulation

in Aged Monkeys
M . T. M U R P H Y A N D J. M . L I P T O N j

Physiology and Anesthesiology Departments, University o f Texas Health Science Center at Dallas
Dallas, TX 75235

R e c e i v e d 18 J u l y 1983

MURPHY, M. T. AND J. M. LIPTON. ~-Endorphin: Effect on thermoregulation in aged monkeys. NEUROBIOL

AGING 4(3) 187-190, 1983.--In previous experiments small doses of the opiate morphine produced greater hyperthermia in
aged than in younger sub-human primates. To test whether this augmented response is due to enhanced sensitivity of CNS
opioid receptors with age,/3-endorphin (0.625-5 tzg), an endogenous opioid peptide, was injected into the lateral cerebral
ventricle (ICV) of young (<9 years) and aged (>9 years) squirrel monkeys. Significantly greater hypertherrnias developed
in the older primates after each dose. In the aged monkeys, all but the smallest dose increased core temperature about 1.5°C
within I hr after injection. Mean rectal temperature in the younger animals rose 0.5--0.7° after all but the largest dose
(I-I.5oc rise). Both groups maintained an elevated body temperature after central /9-endorphin throughout the 5 hr
recording period. 1.25/zg ~8-endorphin given ICV in a hot environment (30"C) caused greater hyperthermia in older animals.
This dose given in the cold (18oc) caused large changes in temperature of the aged monkeys, either hyperthermia or marked
decreases, whereas the young primates developed only moderate rises in body temperature. The same dose of morphine
sulfate 0.25 p.g) ICV produced similar changes in core temperature in the two age groups in each ambient temperature.
These results indicate that: (I) stimulation of CNS opioid receptors influences thermoregulation and (2) aging increases
responsiveness to such stimulation.

/3-Endorphin Thermoregulation Aging Squirrel monkey Hypenhermia

R E C E N T research has shown that certain drugs and endog-
enous substances preferentially promote dysthermia in aged
animals [5, 6, 19, 21, 24, 25]. Thermoregulatory mechanisms
are altered in aged homeotherms, and it may be that en-
hanced sensitivity of the CNS to the temperature-altering
effects of various compounds renders the elderly especially
susceptible to mild dysthermia, accidental hypothermia and
heatstroke [17, 31, 33]. Drugs commonly prescribed for the
elderly, such as chlorpromazine, diazepam and morphine,
and others widely used by this population, including ethanol,
have greater effects on temperature regulation of older lab-
oratory animals [.5, 7, 21, 25]. Certain endogenous CNS pep-
tides, including those derived from proopiomelanocortin
(POMC), affect thermoregulation [14, 19, 24, 36]. The
structurally-related neuropeptides A C T H (I-24) and a - M S H
(!-13) have significant effects on central temperature control
[14, 24, 36]. Central and peripheral injections of a - M S H
lower body temperature [14] and reduce fever [26], effects
that are significantly enhanced in aged homeotherms [ 19,24].
Because systemic and central administration o f small
amounts of morphine sulfate, a drug presumed to exert its
effect via central opioid receptors, produce greater changes
in body temperature o f aged animals [7, 23, 30, 32], the en-
dogenous opioid peptide ~ e n d o r p h i n was tested in the pres-
ent experiments to determine if the thermoregulatory re-
sponse to this POMC derivative is also altered with age in
primates. When age-related changes in core temperature
after/3-endorphin administration were found, the tempera-
ture effects of an intermediate dose were chosen and com-
pared to those caused by morphine in young and older mon-
keys in cold (18 °) and hot (30°C) environments to further
determine the degree o f alteration of CNS opioid sensitivity
with age.
METHOD
Animals
Male and female squirrel monkeys (Saimiri sciureus)
ranging in age from 3--16 + years and 500-1000 g in weight
were used. Birthdates o f these animals were documented,
or, in the case o f some animals over 12 yrs old, the date of
importation was known. The imported animals were known
to be over ! year old at the time o f importation. All animals
were free o f tuberculosis, and only those in good health were
used in the experiments. The animals were individually
caged in a 23--25"C environment with a 12-hr light-dark cycle
and they had lived in the environment for at least 3 months
prior to testing. All experiments were performed during the
|ight cycle and were initiated at approximately the same time
each day. They were maintained on monkey biscuits
(Wayne, Allied Mills, Inc.) and fruit; water was available ad
lib. Before formal testing was begun each monkey was
trained to sit quietly for 8-hr periods in a restraining chair
designed for the species (Piaslab, Inc.). The animals were

JRequests for reprints should be addressed to J. M. Lipton, Physiology Department, Southwestern Medical School, UTHSCD, 5323 Harry
Hines Blvd., Dallas, TX 75235.

187
 188 MURPHY AND LIPTON

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FIG. 1. Hyperthermia (mean change+-S.E.M.) produced by ICV fl-endorphin

(0.625-5 #g) in young (n=6) and older (n=5) squirrel monkeys in a thermoneutral
(25"C) environment.

divided into two age groups: a younger group under nine
years o f age (range: 3-8 years), and an older group aged
10--16+ years. This division at nine years was made to insure
that the responses o f definitely aged monkeys were used in
the comparisons [4].
Surgical Procedure
The monkeys were pre-treated with ketamine hydrochlo-
ride (Vetalar, Parke-Davis; 2-3 mg, IM), atropine sulfate
(Atrosed, Burns-Biotec; 0.02 mg/kg IM) and dexamethasone
(Azium, Schering Veterinary; 0.25 mg/kg, IM) and they were
anesthetized with sodium pentobarbital (Nembutal, Abbott
Laboratories; 15 mg/kg, IP). Each animal was placed in a
K o p f stereotaxic instrument designed for the species, and a
cannula (Kc No. 201) was implanted aseptically into a lat-
eral cerebra_ ventricle (ICV) using coordinates (6.0 mm
anterior, 3.0 mm lateral, vertical until cerebrospinal fluid
appeared in the cannula) derived from the atlas of Emmers
and Akert [1 l]. Dental acrylic was used to secure the cannula
to stainless steel screws driven into the calvarium. Experi-
ments were begun 18-31 days later.
Temperature Measurement and Injections
Monkeys were placed in a restraining chair in an en-
vironmental chamber (Hotpack Corporation) maintained at
25", 18" or 30*(+_I'C, range). A thermistor probe (Yellow
Springs International No. 701) was inserted 10 cm into the
rectum and held in place with tape. Body temperature was
recorded at 10-rain intervals using a digital temperature re-
corder (Digitec Corporation) connected to a MINC I 1 online
computer. Injections were made up daily from a stock solu-
tion and administered after a period of 1 hr or more during
which base-line temperature was recorded. Rectal tempera-
ture was monitored for 5 hr after drug or control injections.
/3-Endorphin (human, Boehringer Mannheim Biochemicals)
and morphine sulfate (MS, Mallinckrodt) were dissolved in
non-pyrogenic isotonic saline and injected ICV (50/11 vol-
ume + 20/zl saline flush). The doses were administered to
each monkey in randomly assigned order. All injections were
separated by at least 72 hr to avoid development of
tolerance.
Calculations
The Thermal Response Index (TRI), the area under the
temperature curve expressed as A°C x hr [9], was used to
quantify the effects o f fl-endorphin and morphine on body
temperature. A N O V A and Student's t-tests were performed
on TRI~ scores, the temperature response for two hours after
injection.
RESULTS
/~-Endorphin (0.625-5/~g) injected ICV in a thermoneutral
environment increased rectal temperature of both young and
old monkeys. Old animals developed a maximum rise in
body temperature of at least 1.5°C after all but the smallest
(0.625 /~g) dose of fl-endorphin, and maintained core tem-
perature at least 1° above baseline throughout the 5 hr re-
cording period (Fig. 1). Young animals also developed long-
term hyperthermias o f approximately 0.6 ° after all but the
largest (5/~g) dose, which caused an approximate 1.2°C rise
in rectal temperature. The increases in body temperature in
the older monkeys were significantly greater (p<0.05 or bet-
ter) than those in the younger group after each dose. There
was no significant difference in pre-injection baseline tem-
peratures o f young (38.22-+0.40 °, S.E.M.) and old
(38.43 +-0.47°C) animals.
~8-ENDORPHIN AND THERMOREGULATION
An intermediate dose (I.25/J,g) W a s selected to test in cold
( 18°) and hot (30°) environments. This dose was chosen since
in a thermoneutral environment it produced a maximum rise
in core temperature in the aged animals, while causing only a
slight increase in body temperature in the younger animals.
Hence any increased sensitivity with age might be evident
under thermal stress at this dose. ICV administration of 1.25
/~g /3-endorphin in the cold caused either hypothermia or
hyperthermia in the older animals. In contrast, younger
animals developed moderate rises in core temperature after
this dose of/3-endorphin (p<0.04, Fig. 2). In the heat, hyper-
thermia was enhanced in the older monkeys compared with
the younger group (p <0.03). The same dose of MS (1.25/~g)
given ICV produced similar changes in body temperature in
each age group at each ambient temperature (Fig. 2). Mean
(_+S.E.M.) baseline temperature of the two groups was not
significantly different in the cold (young--38.30__+0.34°;
oid--38.06_+0.68 °) or in the heat (young--38.38_+0.30°;
oid---38.61_+.+0.48°C). Vehicle injection did not change rectal
temperature of either age group in either ambient tempera-
ture.
No marked behavioral changes were observed in either
age group after /3-endorphin or MS administration in the
three ambient temperatures.
DISCUSSION
Central administration of/3-endorphin altered body tem-
perature in young and aged sub-human primates with the
greatest effects in the aged monkeys. In response to each
dose of/3-endorphin given in a thermoneutral environment
both young and older groups developed significant rises in
body temperature. Aged monkeys developed a maximum
rise after administration of 1.25 p.g whereas in the younger
animals 5/zg was required to elicit a comparable hyperther-
mia. The aged primates also developed significantly greater
dysthermia when 1.25/~g of the opioid peptide was given in
hot and cold environments. In 30°C they developed greater
hyperthermia. Both hyperthermia and hypothermia occurred
in the aged animals after central injection of/3-endorphin in
the cold. The younger animals developed only moderate
rises in body temperature in each environment. A selected
dose of morphine (1.25/zg), an opiate previously shown to
alter normal thermoregulation when given centrally and pe-
ripherally [7, 23, 30, 32], had effects on temperature that
were similar to those after/3-endorphin [2, 18].
In previous research [7] age-related differences in thermal
response were observed in squirrel monkeys after small
doses of morphine sulfate. In this earlier study a weak but
statistically significant correlation between maximum tem-
perature change and age was noted after the smallest sub-
cutaneous dose was given (0.5 mg/kg). For 2.5 and 5.0/~g
doses of morphine given centrally, the positive correlation
between temperature and age were significant, and smaller
doses (0.625 and 1.25 p.g) consistently caused increases in
temperature of monkeys 8.5--16 years old, but only slight
rises in body temperature in the young animals. The
age/temperature relations were marked by increased varia-
bility after larger doses of morphine given either centrally or
peripherally. Thus it appears that the age-related hyper-
thermia caused by/3-endorphin in the present experiments is
more like the effect noted after low doses of morphine in
prior research on this species. The mixed hypotherma/
hyperthermia observed in the aged animals after both
/3-endorphin and morphine in the cold was not expected and
189
To, 25"C 1"= • ~ O ' C ¥o, W°C
~s
4o
! o5
~ oo
~.~/49 ICV
i -to I--"I < 9 I~
:'91"
~ -20
-~c
FIG. 2. Changes in rectal temperature (mean_+S.E.M.) produced by
1.25 p.g/3-endorphin and morphine sulphate ICV in thermoneutral
(25°), hot (30°) and cold (18°C) environments in young (n=6) and
older (nffi6) squirrel monkeys. At 18°C older primates developed
either hyperthermia (n=2) or hypothermia (n=4).
may be related to the "biphasic" temperature effect of mor-
phine described previously [2,8]. In certain aged primates
subjected to cold, intermediate doses of opiate substances
appear to have the same thermal effect (hypothermia) as
larger doses of morphine in a thermoneutral environment.
The hypothermia after large doses of morphine is believed to
reflect a general depression of thermoregulation [8].
The enhanced response of the older monkeys to stimula-
tion of opioid receptors by fl-endorphin may reflect a greater
central responsiveness to this peptide with age, perhaps be-
cause of a change in the opioid receptor population. Since it
has been shown that/3-endorphin levels within the hypothal-
amus decline with age [1,13], it may be that the enhanced
thermal response results from increased sensitivity, perhaps
similar to denervation supersensitivity, of the aged CNS to
this neuropeptide. Alternatively, the increased response may
be due to a more general decrease in CNS temperature con-
trols with aging, resulting in a failure to counter rises in
temperature caused by fi-endorphin. Both explanations are
tenable.
It is not yet clear whether central/3-endorphin has a role
in normal control of thermoregulatory processes. Research
on rats [2, 15, 29] and cats [I0] indicates that the opioid
antagonist naloxone has little or no effect on normal body
temperature. However, ~-endorphin is found in significant
concentrations in brain regions that are important to tem-
perature control [3, 12, 16, 34] and direct injection of this
peptide into the primary temperature control in the preop-
tic/anterior hypothalamic region causes hyperthermia [2, 22,
28]. Although there are questions about the role of central
/3-endorphin in thermoregulation, the age difference in ther-
mal response to this peptide is striking. Consequently, any
drug that influences opiate receptors in the aged brain might
be expected to cause dysthermia, whether the endogenous
opioid peptides are significant to normal thermoregulation or
not. If opioid peptides do have a role in central temperature
control, changes with age in concentration, activity and/or
metabolism might underlie certain age-related modifications
190
in temperature regulation and lead to thermolability, heat
stroke and accidental hypothermia, conditions that are more
c o m m o n in the aged [17, 31, 33].
D e c r e a s e d febrile responses in aged rabbits, squirrel
m o n k e y s and man [4, 20, 27, 35], and age-related reduction
in prostaglandin-induced hyperthermia [20], have been
documented. The diminished responses to pyrogens and pros-
taglandins in the previous research might simply reflect an
inability o f the aged animals to develop and maintain significant
rises in core temperature due to reductions in peripheral ther-
moeffector capacity. H o w e v e r , the present research demon-
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MURPHY AND LIPTON
strates that aged subhuman primates are capable of developing
and sustaining marked rises in body temperature. The reduced
responses to pyrogens and hyperthermic agents observed in
previous experiments may not be the result of diminished pe-
ripheral capacity to produce and c o n s e r v e heat, but rather
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ACKNOWLEDGEMENTS
This research was supported by National Institute on Aging grant
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