Regulatory Peptides 104 (2002) 47 – 53

www.elsevier.com/locate/regpep

Effects of orexins on energy balance and thermoregulation

Miklós Székely a,*, Erika Pétervári a, Márta Balaskó a, István Hernádi b, Boglárka Uzsoki b
a
Department of Pathophysiology, Faculty of Medicine, University of Pécs, 12 Szigeti ut, Pécs, H-7624, Hungary
b
Department of Zoology and Neurobiology, Faculty of Science, University of Pécs, Pécs, Hungary

Abstract

Intracerebroventricular injections of 10 – 20-mg orexin-A induce food intake in rats for about 30 min, or enhance fasting-induced
hyperphagia. In thermoregulatory studies, an amount of 2 mg of the peptide causes hypometabolism and hypothermia in the same period. The
thermoregulatory reaction can be demonstrated at moderately cool environments, mainly after slight food deprivation. Both the ingestive and
the thermoregulatory reactions are more pronounced in cold-adapted animals. Pretreatment with D-Tyr27,36,D-Thr32-NPY(27 – 36), a peptide-
antagonist of NPY, prevents the hypothermia. It is concluded that, probably through NPY activation, orexin-A is involved primarily in the
regulation of energy status of the body (as an anabolic agent), and not simply in the regulation of either food intake or body temperature. This
anabolic response is followed by a late and more sustained catabolic phase characterized by absence of food intake, increased metabolism and
dose-dependent hyperthermia, which hyperthermia cannot be suppressed by the NPY-antagonist. In contrast to orexin-A, neither hyperphagia
nor suppression of refeeding hyperphagia, but dose-dependent hyperthermia follows injections of orexin-B, suggesting that this peptide has
neither coordinated anabolic nor coordinated catabolic effects on energy balance. D 2002 Elsevier Science B.V. All rights reserved.

Keywords: Food intake; Fasting; Operant behavior; Metabolic rate; Temperature; Neuropeptide Y

1. Introduction run, demands of metabolic rate must be met by proportional

Neuropeptides have long been known to participate in
the regulation of food intake, body weight and metabolic
processes [1]. Various studies of the past few years have
established an important place for orexins (hypocretins) of
the lateral hypothalamic area among the central orexigenic
substances [2– 6]. Exogenous (centrally injected) orexins
have been demonstrated to induce food intake [7 – 9], while
hypothalamic levels of prepro-orexin mRNA became ele-
vated after fasting [10,11].
Apart from their effects on ingestive behavior, orexins
have also been reported to participate in, or have an in-
fluence on, various physiological processes such as the re-
gulation of cardiovascular functions [12 – 14], behavior
[15 – 17], sleep/wake cycle [17,18], endocrine functions
[19], and they may also influence metabolic rate [12,20]
and the regulation of body temperature [21].
Although in studies using orexigenic peptides the regu-
lation of food intake is by far the most frequently investigated
aspect of energy balance, these studies cover only one side of
the problem. Maintenance of energy homeostasis depends on
long-term and short-term regulatory processes. In the long
(although periodical) food intake; the abnormalities result in
changes of body weight. The short-term balance between
energy production (actual metabolic rate) and energy dis-
sipation is a function of temperature regulation; any imbal-
ance is manifested as change in body temperature.
In this paper an attempt is made for integrated analysis of
the effects of orexins on energy status in the rat, including
those effects they have on ingestive behavior and on
thermoregulation. The effects of orexin-A and orexin-B
might not be identical [7 –9,20]. The observed orexin effects
are to be compared with those described for neuropeptide Y
(NPY) because some orexin actions resemble the effects of
NPY [21] and an interaction is possible between the two
peptides [5,23 – 25]. Besides, since thermal adaptation may
modify both food intake and thermoregulatory reactions
(energy status, in general) [22,26,27], orexin actions were
compared in control non-adapted and cold-adapted rats.
2. Materials and methods
2.1. Animals

*
Corresponding author. Tel.: +36-72-536246; fax: +36-72-536247. Adult Wistar rats of both sexes were used from the colony
E-mail address: miklos.szekely@aok.pte.hu (M. Székely). of the Department of Pathophysiology. The animals were

0167-0115/02/$ - see front matter D 2002 Elsevier Science B.V. All rights reserved.
PII: S 0 1 6 7 - 0 11 5 ( 0 1 ) 0 0 3 4 8 - 2
48 M. Székely et al. / Regulatory Peptides 104 (2002) 47–53
maintained singly in plastic boxes with some woodchip
bedding, where conventional rat chow and water were
available ad libitum. All rats were habituated to handling
and daily weighing procedures. Besides, they were accus-
tomed to a semi-restraining cage, which prevented them from
turning around, without blocking their other movements—
these animals were used in studying temperature regulation
and metabolic rate, or in observing body weight changes in
the course of feeding. The temperature of the animal room
was either 22 – 25 C (non-adapted group, NA) or 3 – 5 C
(cold-adapted group, CA) for a period of at least a month, with
lights on between 06:00 and 18:00 in both rooms. Some other
NA rats were accustomed to a Colbourn Habitest system,
which enabled us to study operant behavior in food ingestion.
Most animals underwent several (feeding, thermoreg-
ulation) tests: altogether 69 rats were used on 231 occasions.
After finishing the last tests, all rats were given a narcotic
overdose. Location of the implanted cerebroventricular
cannula was checked macroscopically at postmortem exami-
nation, and only data of those rats were accepted in which
the right location was confirmed. All experiments and in-
terventions were undertaken according to the general rules
and special approval of the University of Pécs Ethical Com-
mittee for the Protection of Animals in Research.
2.2. Surgery, injection methods
At the end of the adaptation period and 1 week before
the first testing, a 22-gauge stainless steel guide cannula
was implanted under intraperitoneal ketamine (Calypsol,
Richter) + xylazine (Rometar, Spofa) (78 + 13 mg/kg, respec-
tively) anesthesia into a lateral cerebral ventricle (intracere-
broventricularly, ICV; A: 1.0, L: 1.5, V: 3.8 mm). On test-
days an injection cannula was inserted. This was connected
to a polyethylene tube containing the required amount of
orexin-A or orexin-B (Bachem) in a volume of 5 ml, or a
combination of the NPY-antagonist D-Tyr27,36, D-Thr32-
NPY(27 –36) (Bachem, referred to as H-3328) plus orexin-
A, both in 5 ml. Similar volume of the solvent (0.9% NaCl)
was injected in control experiments. Small bubbles separated
the active substances from each other or from the physio-
logical saline that filled up the rest of the 25 –30-cm-long
tube. This length served to give injections remotely. Most
rats were used repeatedly in various tests, but at least 1 week
elapsed between two tests with ICV injections. Injections
were given from around 10:00 to 12:00.
2.3. Assessment of food intake
Two methods were used for testing the effect of peptides
on food intake, both on female rats. According to the first
one, the operant behavior was analyzed by a Colbourn
Habitest system (Colbourn Instruments, Allentown, PA):
upon pushing its head into the feeder, the rat interrupted a
photocell system and a pellet was released to the feeder. The
pellets were of dustless precision Bioserve PLT20 variety,
45 mg each. The released pellets were counted continuously.
For 3 consecutive days prior to the test, the animals were
trained to use the system and they never left uneaten pellets
in the feeder. Consumption of pellets was measured after
ICV injections of orexin-A or 0.9% NaCl and the cumu-
lative values were given at 20-min intervals.
According to the second method, the unanesthesized rats
received an ICV injection of orexin or 0.9% NaCl and 10
min later the conventional chow was offered in their home
cage. In this case, the body weights were measured every 30
min for a period of 3 h, as well as the cumulative chow
consumption by the end of the whole period. Naturally,
concurrent water intake and urinary/fecal losses also modi-
fied body weights, but the maximal weight changes were
still in good correlation with the cumulative food intake
[27]. Thus, weight changes (expressed as percent of initial
body weight) were accepted as indicators of food intake, and
the course of weight changes gave a rough estimate of the
dynamics of food intake. In some cases, food (but not water)
was withdrawn for 24 h prior to testing, in order to check the
possibility that refeeding hyperphagia is altered by orexins.
2.4. Thermoregulatory tests
For testing thermoregulatory responses, the unanesthe-
sized rats were placed in the semi-restraining cage in which
copper-constantan thermocouples were attached to them for
measuring temperatures of the colon (Tc) and the tail skin
(Ts). Together with this cage, the rats were placed into an
open-circuit brass metabolic chamber which, in turn, was
immersed into a thermostatically controlled water bath of
thermoneutrality (28 – 30 C for NA and 25 C for CA rats)
or 10 C below thermoneutrality (20 C for NA and 15 C
for CA rats). Metabolic rate (MR) was measured by the help
of a Kipp– Noyons diaferometer. Temperature of the cham-
ber (Ta), along with Tc, Ts and MR were continuously
recorded for a period of 3 h following ICV injections. In
order to see the other side of thermal balance, heat loss was
assessed from the heat loss index (HLI = Ts Ta/Tc Ta)
calculated according to earlier methods [21].
2.5. Statistical analysis
One-way ANOVA with Scheffe’s post hoc tests and
Student’s t-tests were used for statistical analyses. Statistical
significance was accepted at the level of p < 0.05. Figures
demonstrate mean values F S.E.M.
3. Results
3.1. Effect of orexins on food intake and body weight
In the operant behavior model, the novelty of the pellet
initiated feeding: normally fed rats started to eat immedi-
ately after being placed into the system, even if they were
 M. Székely et al. / Regulatory Peptides 104 (2002) 47–53 49

Fig. 1. Cumulative number of pellets (FI = food intake) consumed by rats
(n = 4) following ICV injection of 0.9% NaCl (—) or 10 mg orexin-A (.-.).
Operant behavior studied in a Colbourn Habitest system. Asterisks (*):
statistically significant differences between the two tests (Student).
Fig. 3. Rise of BW in 24-h food-deprived CA rats in the course of refeeding
(in percent of fasting BW) after ICV injections of 0.9% NaCl (—, n = 12),
20 mg orexin-A (.-., n = 6), or 20 mg orexin-B (6-6, n = 6). Chow offered
10 min after injection. Asterisk (*) denotes statistically significant
difference (Scheffe).

not injected or were injected ICV with 0.9% NaCl. How-

ever, this was confined to the first 20-min period, except in
rats injected ICV with 10-mg orexin-A in which pellet
consumption continued and the cumulative intake became
significantly higher by the second 20-min period (Fig. 1).
The difference between control and orexin-A-treated ani-
mals did not increase any more in the next periods.
Cumulative 3-h chow consumption also increased after
ICV administration of orexin-A: following 20-mg ICV
orexin-A injection, NA rats consumed 3.56 F 0.47 g chow
(vs. 0.81 F 0.32 g after ICV saline treatment), while in CA
rats similar injection induced more pronounced food intake
(4.33 F 0.63 vs. 1.68 F 0.40 g following ICV saline, or
1.47 F 0.51 g after 20 mg ICV orexin-B). For orexin-A the
response was dose-dependent: smaller doses elicited smaller
food intakes.
Similarly to the differences in chow consumption, greater
body weight rises (reflecting greater food intakes) followed
the orexin-A injections in CA than in NA rats, but the

Fig. 2. Body weight changes (DBW) in percent of initial BW in CA and
NA rats after ICV injection of 0, 2, 10 or 20 mg orexin-A. The initial body
weights were similar in various groups; n = 13, 6, 10, 6, respectively, for
CA, and 9, 8, 7, 8 for NA rats.
Fig. 4. Changes of body temperature (Tc, upper panel) and metabolic rate
(MR, lower panel) in non-adapted rats at 20 C environmental temperature
following ICV injection of 0.9% NaCl (interrupted line, n = 6), or 2 mg
orexin-A (6-6, n = 5). Similar data for 24-h food-deprived rats: ICV
injection of 0.9% NaCl (—, n = 6), or 2 mg orexin-A (.-., n = 6). For clarity,
S.E.M. is not shown for control MR curves. The initial Tc and MR values
were similar in all groups. Asterisk (*): significant difference vs. all other
groups, (x): significant difference between the two orexin-treated groups
(Scheffe). In the interval of 20 – 60-min significant differences between
MR-s of the two orexin-treated groups (Student).
50 M. Székely et al. / Regulatory Peptides 104 (2002) 47–53
effects were dose-dependent in both groups. Fig. 2 demon-
strates the fractional body weight changes within a 90-min
period: the greatest rise was observed in the first 30-min
interval, further weight gains in the consecutive periods
were smaller or negligible. In the first interval clear dose-
dependence is obvious, and also a remarkable difference
between the responses of CA and NA rats.
Fig. 3 shows that orexin-A was even more efficient in
food-deprived CA rats: it strongly enhanced the body
weight rise in the course of refeeding hyperphagia. Ore-
xin-B neither enhanced nor suppressed the refeeding hy-
perphagia.
3.2. Orexin effects on regulation of body temperature
Smaller orexin-A doses were sufficient to affect Tc than
the doses necessary to influence food intake. In these studies
the peptide was administered at moderately cool environ-
Fig. 5. (a) Changes of Tc in CA rats injected ICV with various doses of
orexin-A at an ambient temperature of 15 C. The initial Tc values were
between 37.2 and 37.8 C (n = 6, 6, 9, or 8 for doses of 0, 0.5, 2, or 10 mg
orexin-A, respectively). (b) Changes of Tc in CA rats injected ICV at 15 C
with 0.9% saline (- - -, n = 6), 2 mg orexin-A (.-., n = 13), 10 mg NPY-
antagonist H-3328 (—, n = 9), or orexin-A 10 min after H-3328 (6-6,
n = 7). The initial Tc values varied between 37.6 and 37.8 C. H-3328 given
at arrow. Asterisks (*): significant hypothermia for orexin-A-treated rats
(vs. all others, Scheffe), at 110 – 180-min interval significant hyperthermia
(.-., 6-6, vs. saline-treated controls).
Fig. 6. Changes of Tc in CA rats-injected ICV with 2 mg orexin-A at
thermoneutrality (25 C) (6-6, n = 7) or at 15 C (.-., n = 13). Control CA
rats received 0 mg peptide (0.9% NaCl) at 25 C (interrupted line, n = 6) or
at 15 C (—, n = 5). Initial Tc values: 37.4 – 37.9 C. Asterisk (*): significant
hypothermia (Scheffe) or hyperthermia (Student), diamond (x): significant
hyperthermia (orexin vs. control at 25 C, Scheffe).
ments (20 vs. 15 C for NA vs. CA rats, respectively); not
too cold in order that a Tc-rise could still be manifested by
further MR-rise, and cold enough that a Tc-fall could also
easily develop by suppression of the cold-induced MR-rise.
In NA rats, 2-mg orexin-A ICV injection (which, in CA
rats, evoked maximal responses) induced only a slight, late
Tc-rise (Fig. 4), 1 mg was occasionally followed by an early
Tc-fall, and less peptide was ineffective. However, 2 mg
regularly suppressed MR and Tc in those NA rats, which had
been food-deprived for 24 h prior to the test.
In contrast to NA rats, in CA animals (without food
deprivation) characteristic biphasic Tc-changes followed the
ICV injections of 2-mg orexin-A: an initial Tc-fall lasting for
30 – 40 min was followed by a longer-lasting Tc-rise (Fig.
5a). The hypothermia was maximal and regular at this dose
and was less clear or absent when using smaller or bigger
doses; while the hyperthermic late phase was greater after
larger doses of the peptide. Since HLI was continuously low
(not demonstrated), in the present studies (unlike in our
preliminary data [21]), radiant heat loss did not contribute to
the observed hypothermia. Not heat loss, but a moderate
MR-depression was responsible for the Tc-fall, similarly as
in the fasting NA rats of Fig. 4. In the hyperthermic phase
maximal skin vasoconstriction was coupled with elevated
MR.
In CA rats at moderately cool (15 C) environment, the
orexin-A-induced early hypothermia, but not the late hyper-
thermia, was prevented by ICV pretreatment with NPY-
antagonist H-3328 (Fig. 5b), although the antagonist alone
did not cause any immediate thermoregulatory change (it
was followed by a late, slight hyperthermia).
If the peptide was given at thermoneutrality, no MR-fall
or HLI-rise and no hypothermic phase were seen (Fig. 6);
only the late hyperthermic effect was observed.
Interestingly, the metabolic and thermal responses to
orexin-B were qualitatively different from the orexin-A-
induced responses in that, at any dose applied, orexin-B
 M. Székely et al. / Regulatory Peptides 104 (2002) 47–53
Fig. 7. Changes of Tc in CA rats injected ICV with 0, 2, 10, or 20 mg orexin-
B at an ambient temperature of 15 C (interrupted line, —, 6-6, or .-.,
respectively, with n = 6, 6, 5, or 4). The initial Tc values were between 37.5
and 37.9 C.
elicited hyperthermia only (Fig. 7), never hypothermia.
Particularly pronounced early hyperthermia was seen upon
injection of large orexin-B dose, which was followed by a
second, late Tc-rise.
4. Discussion
Confirming earlier reports [2,3,5,9], the present data show
that orexin-A, rather than orexin-B [7,8,20,28], can induce
food intake in satiated animals. It is a new finding wherein
thermal adaptation modifies the peptide action; following
identical ICV orexin-A doses, CA rats ate more and gained
significantly more weight than their NA counterparts. This is
in line with previous findings that cold-adaptation can
increase the responsiveness to centrally applied prostaglan-
din or NPY [27]. Alternatively, the clearly hyperphagic CA
rats might be assumed to have continuously high orexigenic
activity [22] and to be more hungry at the time of injection.
Fasting also increases orexigenic activity and it enhances the
ingestive response to orexin-A (Fig. 3).
Figs. 1 and 2 demonstrate that the feeding activity eli-
cited by a single orexin-A injection is limited to a period of
about 30 – 40 min. This is in accord with earlier studies in
which orexin-A induced only transient, if any, elevations in
food intake [25] even in the course of chronic ICV admin-
istration [15,29] and, unlike NPY [30,31], this peptide could
not cause obesity [28,29]. A special orexin receptor antag-
onist was still able to suppress normal nocturnal feeding
[32], and anti-orexin serum attenuated fasting-induced food
intake [25,33]. Apparently, orexin-A might participate in
mechanisms of normal (not exaggerated) food intake and
normal energy homeostasis. Small ICV doses of orexin-A
delayed satiety, larger doses caused sedation followed by a
behavioral sequence of eating, grooming, locomotion, sniff-
ing, etc. [34].
The orexigenic effect of orexin-A, although weaker,
resembles the effect of NPY of the arcuate nucleus [3,35 –
51
37]. An interaction of the two peptides has also been
suggested: NPY antagonism successfully prevented the
ingestive response to central orexin-A administration
[5,23 – 25]. Following ICV orexin-A injection, a high c-fos
activity was most strongly expressed in the paraventricular
and the arcuate nuclei [8,38], which nuclei are involved also
in NPY actions.
Food intake responses to orexins, although very impor-
tant, describe only a part of the role these peptides play in
energy homeostasis: food intake and MR determine the
long-term balance. The present and earlier data [20,21]
show that the short-term balance of energy homeostasis
(MR/thermoregulation) is also affected by orexins. The
question arises whether the two (feeding and metabolic/
thermal) actions are merely coinciding or are parts of an
integrated regulatory pattern, which regulation might par-
ticipate in complex adaptive physiological responses. The
pattern may be either anabolic, i.e., increase/retention of
body energy content (tendency for enhanced food intake
plus MR-depression and consequent falls in Tc and heat
loss), or catabolic, i.e., decrease of energy content (satiety
with high MR and Tc). Fasting versus postprandial states
might be appropriate examples [26]. Some substances may
influence food intake and MR independently, and not in an
integrated fashion (e.g. neurotensin may cause hypometab-
olism plus hypophagia; thyroxine induces hypermetabolism
with high food intake); these substances modify energy
content but cannot be regarded as primary regulators of
energy balance. Accordingly, the food intake-modulating
actions of orexins ought to be related to their metabolic/
thermoregulatory actions, to see whether or not these pep-
tides can be accepted as such regulators of energy status.
Similar to the effects on ingestion, the thermoregulatory
effects of orexin-A also resemble the effects of NPY [21] and
may be characterized as a biphasic reaction: a short-lived
initial hypometabolism/hypothermia is followed by a longer-
lasting late hyperthermic period. As shown for ICV serotonin
[39], the fast thermoregulatory reactions might be more
specific than the late ones. Thus, the orexin-induced early
hypothermia, rather than the late hyperthermia, is the effect
which should be regarded a relatively specific action (though
not necessarily a direct one). However, the hypothermia can
be demonstrated (1) only with certain doses, (2) only at
certain (cool) ambient temperatures, (3) only in CA or fasting
NA rats—these findings would need some explanation.
(Ad 1) In the present experiments, a narrow bell-shaped
(not uncommon) dose – response relationship was found for
the orexin-induced hypothermia, with maximal and very
consistent hypothermic effect at a dose of 2 mg. It is not a
unique finding that smaller doses were sufficient to influ-
ence Tc than those needed to induce hyperphagia: similar
differences were found for the effects of NPY [our unpub-
lished data].
(Ad 2) Without enhanced heat loss, manifestation of any
hypothermic action is rather difficult at a thermoneutral
environment: at this temperature MR (being at its minimal
52 M. Székely et al. / Regulatory Peptides 104 (2002) 47–53
level) can decrease only under the influence of toxic
substances. In the cold, however, already minor suppression
of the cold-induced MR-elevation can cause hypothermia.
Since orexin-A is not toxic, it does not enhance heat loss,
but can suppress cold-induced MR-rise; the orexin-induced
temperature fall may be only mild or completely missing at
thermoneutrality, but may be expressive at cool environ-
ments.
(Ad 3) Enhanced activity of orexigenic peptides is likely
to participate in the development of spontaneous hyper-
phagia in CA animals [22,37]. Food deprivation also acti-
vates central orexigenic mechanisms [3,5,6]. These possibly
explain our finding that orexin-A caused hypometabolism
with hypothermia (and marked food intake) in CA rats and
in fasting NA ones, more than in well-fed NA animals. At
the same time, this is a spectacular demonstration of the
difficulties of interpreting thermoregulatory responses or,
eventually, the lack of such responses.
It is worth emphasizing that the period of hypometabo-
lism/hypothermia was in good correlation with the period of
increased food intake (Fig. 2 vs. Fig. 5). Thus, these effects
of orexin-A appear to form a coordinated energy-saving
(anabolic) regulatory pattern, resulting in an increase of
energy content of the body both by increased food intake
and by depressed metabolism.
The orexin-A effects are not necessarily direct actions. In
the orexigenic effects, a role has been suggested for NPY
[5,23 –25]: various NPY-antagonizing methods antagonized
the food intake response to centrally administered orexin-A.
In the present experiments (Fig. 5b) the NPY-antagonist H-
3328 prevented the hypothermic phase of the thermal
response to orexin-A, without affecting the late hyperther-
mia. Both the hypothermia-inducing and the orexigenic
effects of orexin-A (unlike the phenomena of the late
hyperthermia) are NPY-dependent, reinforcing our conclu-
sion that the effects are connected as parts of an integrated
anabolic regulatory pattern.
In mice, ICV-injected orexin-A was reported to induce
feeding, and also a rise in MR [20]. From the point of view
of energy balance, this appears to be rather controversial:
food intake increases energy content in the body, while the
MR-rise acts in the opposite direction, it decreases energy
content. These data seem to be at variance with our findings
in rats.
The later phases of orexin-A action, with hypermetabo-
lism and hyperthermia, are in contrast to the early regulatory
effects of the peptide. Apparently, Tc-rise is the more
common (though probably less specific) consequence of
orexin-A injections. It must be emphasized, however, that
this phase sets in relatively late, at a time when the peptide
effect on food intake is already wearing off. Altogether these
seem to suggest the presence of an integrated energy-
utilizing/wasting (catabolic) regulatory mode, just the oppo-
site of that seen in the early phase. It is also possible that the
grooming behavior [34] is related to this phase: grooming is
not simply a behavioral phenomenon but, as saliva spread-
ing, it is also a characteristic mechanism of heat-dissipation
in cases of hyperthermia [40]. Despite the dose-dependence
of the late hyperthermia, it is probably also an indirect effect
of orexin-A. For example, the anorexigenic CRH has also
been suggested to play a role in conveying some orexin-A
effects [19,25] and other studies [41] have indicated CRH to
increase MR and induce hyperthermia.
Orexin-B appears to differ from orexin-A in that orexin-
B is involved mainly in behavioral and other processes
[15,42] and only less intimately in the energy status or
normal food intake [43]. This peptide was found either
ineffective [7,28] or only slightly effective [8,9] in inducing
ingestive behavior. In the present experiments, orexin-B did
not elicit food intake, nor did it enhance refeeding hyper-
phagia (Fig. 3). The effects of orexin-B appear to be dif-
ferent also in thermoregulation: exclusively hyperthermia
followed the administration of the peptide (Fig. 7). With
smaller doses this started late, but in case of large dose this
was preceded by an immediate, sharp first-phase rise in
body temperature. The hyperthermic effect was not accom-
panied by anorexia: orexin-B did not attenuate refeeding
hyperphagia (Fig. 3). Apparently, no consistent anabolic or
catabolic pattern can be demonstrated for orexin-B, and this
substance probably has some other roles.
In conclusion, it is suggested that orexin-A is not in-
volved primarily or separately in the regulation of either
food intake or body temperature, rather it is a regulator of
the energy status of the body. Its specific effect is likely to
be improvement of energy status by increasing food intake
and decreasing energy wastage. This is a short-lived imme-
diate anabolic effect, which is followed by a late and more
sustained catabolic phase characterized by absence of food
intake and enhanced metabolism. Despite the fact that
orexin-B can also influence body temperature, this peptide
does not seem to be of primary importance in the regulation
of energy balance.
Acknowledgements
The authors are grateful for the most reliable and
conscientious technical assistance of Ms. M. Koncsecskó-
Gáspár, Ms. E. Pákai and Ms. A. Bóka-Kis. The valuable
critical comments of Prof. Z. Szelényi are greatly appre-
ciated. This work was supported in part by the National
Scientific Grant Agency of Hungary (OTKA T026511).
References
[1] Morley JE. Neuropeptide regulation of appetite and weight. Endocr
Rev 1987;8:256 – 87.
[2] Sakurai T. Orexins and orexin receptors: implication in feeding be-
havior. Regul Pept 1999;85:25 – 30.
[3] Kalra SP, Dube MG, Pu S, Xu B, Horvath TL, Kalra PS. Interacting
appetite-regulating pathways in the hypothalamic regulation of body
weight. Endocr Rev 1999;20:68 – 100.
[4] Shiraishi T, Oomura Y, Sasaki K, Wayner MJ. Effects of leptin and
 M. Székely et al. / Regulatory Peptides 104 (2002) 47–53
orexin-A on food intake and feeding related hypothalamic neurons.
Physiol Behav 2000;71:251 – 61.
[5] Williams G, Harrold JA, Cutler DJ. The hypothalamus and the regu-
lation of energy homeostasis: lifting the lid on a black box. Proc Nut
Soc 2000;59:385 – 96.
[6] Schwartz MW, Woods SC, Porte Jr D, Seeley RJ, Baskin DG. Central
nervous system control of food intake. Nature 2000;404:661 – 71.
[7] Dube MG, Kalra SP, Kalra PS. Food intake elicited by central admin-
istration of orexins/hypocretins: identification of hypothalamic sites of
action. Brain Res 1999;842:473 – 7.
[8] Edwards CM, Abusnana S, Sunter D, Murphy KG, Ghatei MA,
Bloom SR. The effect of the orexins on food intake: comparison with
neuropeptide Y, melanin-concentrating hormone and galanin. J Endo-
crinol 1999;160:R7 – 12.
[9] Sweet DC, Levine AS, Billington CJ, Kotz CM. Feeding response to
central orexins. Brain Res 1999;821:535 – 8.
[10] Cai XJ, Widdowson PS, Harrold J, Wilson S, Buckingham RE, Arch
JRS, et al. Hypothalamic orexin expression: modulation by blood
glucose and feeding. Diabetes 1999;48:2132 – 7.
[11] Yamamoto Y, Ueta Y, Serino R, Nomura M, Shibuya I, Yamashita H.
Effects of food restriction on the hypothalamic prepro-orexin gene
expression in genetically obese mice. Brain Res Bull 2000;51: 515 –
21.
[12] Peyron C, Tighe DK, van den Pol AN, de Lecea L, Heller HC,
Sutcliffe JG, et al. Neurons containing hypocretin (orexin) project
to multiple neuronal systems. J Neurosci 1998;18:9996 – 10015.
[13] Samson WK, Gosnell B, Chang JK, Resch ZT, Murphy TC. Cardio-
vascular regulatory actions of the hypocretins in brain. Brain Res
1999;831:248 – 53.
[14] Shirasaka T, Nakazato M, Matsukura S, Takasaki M, Kannan H.
Sympathetic and cardiovascular actions of orexins in conscious rats.
Am J Physiol 1999;277:R1780 – 5.
[15] Ida T, Nakahara K, Katayama T, Murakami N, Nakazato M. Effect of
lateral cerebroventricular injection of the appetite-stimulating neuro-
peptide, orexin and neuropeptide Y, on the various behavioral activ-
ities of rats. Brain Res 1999;821:526 – 9.
[16] Hagan JJ, Leslie RA, Patel S, Evans ML, Wattam TA, Holmes S, et al.
Orexin A activates locus coeruleus cell firing and increases arousal in
the rat. Proc Natl Acad Sci U S A 1999;96:10911 – 6.
[17] Estabrooke I, McCarthy MT, Ko E, Chou TC, Chemelli RM, Yana-
gisawa M, et al. Fos expression in orexin neurons varies with behav-
ioral state. J Neurosci 2001;21:1656 – 62.
[18] Kilduff TS, Peyron C. The hypocretin/orexin ligand-receptor system:
implications for sleep and sleep disorders. Trends Neurosci 2000;
23:359 – 65.
[19] Jászberényi M, Bujdosó E, Pataki I, Telegdy G. Effects of orexins on
hypothalamic-pituitary-adrenal system. J Neuroendocrinol 2000;
12:1174 – 8.
[20] Lubkin M, Stricker-Krongrad A. Independent feeding and metabolic
actions of orexins in mice. Biochem Biophys Res Commun 1998;
253:241 – 5.
[21] Balaskó M, Szelényi Z, Székely M. Central thermoregulatory effects
of neuropeptide Y and orexin A in rats. Acta Physiol Hung 1999;
86:219 – 22.
[22] Székely M, Balaskó M. Hyperphagia in cold-adapted rats: a possible
role for neuropeptide Y. Acta Physiol Hung 1999;86:287 – 91.
[23] Dube MG, Horvath TL, Kalra PS, Kalra SP. Evidence of NPY Y5
receptor involvement in food intake elicited by orexin A in sated rats.
Peptides 2000;21:1557 – 60.
[24] Yamanaka A, Kunii K, Nambu T, Tsujino N, Sakai A, Matsuzaki I, et
al. Orexin-induced food intake involves neuropeptide Y pathway.
Brain Res 2000;859:404 – 9.
53
[25] Ida T, Nakahara K, Kuroiwa T, Fukui K., Nakazato M, Murakami T, et
al. Both corticotropin releasing factor and neuropeptide Y are involved
in the effect of orexin (hypocretin) on the food intake in rats. Neurosci
Lett 2000;293:119 – 22.
[26] Székely M. The vagus nerve in thermoregulation and energy metab-
olism. Auton Neurosci Basic Clin 2000;85:26 – 38.
[27] Uzsoki B, Fekete Á, Pétervári E, Balaskó M, Székely M. Enhanced
responsiveness to central prostaglandin E or neuropeptide Y in cold-
adapted rats. J Therm Biol 2001;26:519 – 23.
[28] Haynes AC, Jackson B, Overend P, Buckingham RE, Wilson S, Ta-
dayyon M, et al. Effects of single and chronic intracerebroventricular
administration of the orexins on feeding in the rat. Peptides 1999;20:
1099 – 105.
[29] Yamanaka A, Sakurai T, Katsumoto T, Yanagisawa M, Goto K.
Chronic intracerebroventricular administration of orexin-A to rats in-
creases food intake in daytime, but has no effect on body weight.
Brain Res 1999;849:248 – 52.
[30] Stanley BG, Kyrkouli SE, Lampert S, Leibowitz SF. Neuropeptide Y
chronically injected into the hypothalamus: a powerful neurochemical
inducer of hyperphagia and obesity. Peptides 1986;7:1189 – 92.
[31] Zarjevski N, Cusin I, Vettor R, Rohner-Jeanrenaud F, Jeanrenaud B.
Chronic intracerebroventricular neuropeptide-Y administration to nor-
mal rats mimics hormonal and metabolic changes of obesity. Endo-
crinology 1993;133:1753 – 8.
[32] Haynes AC, Jackson B, Chapman H, Tadayyon M, Johns A, Porter
RA, et al. A selective orexin-1 receptor antagonist reduces food con-
sumption in male and female rats. Regul Pept 2000;96:45 – 51.
[33] Yamada H, Okumura T, Motomura W, Kobayashi Y, Kohgo Y. In-
hibition of food intake by central injection of anti-orexin antibody in
fasted rats. Biochem Biophys Res Commun 2000;267:527 – 31.
[34] Rodgers RJ, Halford JC, Nunes de Souza RL, Canto de Souza AL,
Piper DC, Arch JRS, et al. Dose – response effects of orexin-A on food
intake and the behavioural satiety sequence in rats. Regul Pept
2000;96:71 – 84.
[35] Billington CJ, Briggs JE, Grace M, Levine AS. Effects of intracere-
broventricular injection of neuropeptide Y on energy metabolism. Am
J Physiol 1991;260:R321 – 7.
[36] Bouali M, Fournier A, St-Pierre S, Jolicoeur FB. Effects of NPY and
NPY2 – 36 on body temperature and food intake following administra-
tion into hypothalamic nuclei. Brain Res Bull 1995;36:131 – 5.
[37] Bing C, Frankish HM, Pickavance L, Wang Q, Hopkins DF, Stock
MJ, et al. Hyperphagia in cold-exposed rats is accompanied by de-
creased plasma leptin but unchanged hypothalamic NPY. Am J Phys-
iol 1998;274:R62 – 8.
[38] Mullett MA, Billington CJ, Levine AS, Kotz CM. Hypocretin I in the
lateral hypothalamus activates key feeding-regulatory brain sites. Neu-
roReport 2000;11:103 – 8.
[39] Székely M. 5-hydroxytryptamine induced changes in body temper-
ature of newborn kittens and guinea-pigs and the effect of indometha-
cin thereon. Experientia 1978;34:58 – 9.
[40] Hart JS. Rodents. In: Whittow GC, editor. Comparative Physiology of
Thermoregulation, vol. II. New York: Academic Press; 1971. p. 1 –
149.
[41] Rothwell NJ, Stock MJ. Central effects of CRF on metabolism and
energy balance. Neurosci Biobehav Rev 1989;14:263 – 71.
[42] Jones DN, Garlton J, Parker F, Taylor SG, Routledge C, Hemmati P, et
al. Effects of centrally administered orexin-B and orexin-A: a role for
orexin-1 receptors in orexin-B-induced hyperactivity. Psychopharma-
cology 2001;153:210 – 8.
[43] Cai XJ, Denis R, Vernon RG, Clapham JC, Wilson S, Arch JRS, et al.
Food restriction selectively increases hypothalamic orexin-B levels in
lactating rats. Regul Pept 2001;97:163 – 8.