European Journal of Clinical Investigation (2002) 32, 236 – 241

Histidine induces lipolysis through sympathetic nerve in

Blackwell Science Ltd

white adipose tissue

H. Yoshimatsu*, K. Tsuda*, A. Niijima†, M. Tatsukawa*, S. Chiba* and T. Sakata*

*
Oita Medical University, Oita, Japan and †Department of Physiology, Niigata University, School of Medicine,
Niigata, 951 Japan

See commentary on page 221.

Abstract Background Hypothalamic neuronal histamine has been shown to increase lipolysis in white
adipose tissue. The present study aimed to clarify whether peripheral loading with L-
histidine, a precursor of neuronal histamine, may affect lipid metabolism in adipose tissue.
Materials and methods The in vivo microdialysis study was used to assess lipolysis in rat
epididymal adipose tissue by measuring the release of glycerol in response to administration
of L-histidine. In addition, electrophysiological measurements were performed to record
changes in activity of sympathetic nerve innervating adipose tissue following histidine
treatment.
Results Sequential administration of isoproterenol, a β-adrenoceptor agonist, through the
microdialysis cannula at concentrations of 10–8 to 10–6 M increased the glycerol
concentration in the dialysate dose-dependently (P < 0·05). Intraperitoneal administration
of L-histidine at a dosage of 0·35 mmol kg–1 also increased the glycerol concentration
compared to that of phosphate buffered saline (P < 0·05). Concomitantly, the administration
of histidine increased the serum concentration of free fatty acid compared to control
treatment (P < 0·05). The accelerating effects of histidine on lipolysis were mimicked by
the infusion of 102 nmol rat–1 L-histamine into the third cerebroventricle (P < 0·05).
Electrophysiological measurement demonstrated that administration of histidine at a dosage
of 0·35 mmol kg–1 increased the activity of efferent sympathetic nerve, innervating adipose
tissue more than the infusion of phosphate buffered saline (P < 0·05).
Conclusion The present results indicate that histidine accelerates lipolysis in white adipose
tissue through activation of the sympathetic nerve. The regulation of lipolysis may therefore
involve histamine neurons in the brain, probably through the conversion of L-histidine to
histamine in the hypothalamus.
Keywords Epididymal adipose tissue, in vivo microdialysis, L-histidine, lipolysis, neuronal
histamine, sympathetic nerve.
Eur J Clin Invest 2002; 32 (4): 236–241

Introduction

Activation of hypothalamic histamine neurons has been

Department of Internal Medicine I, School of Medicine, Oita
Medical University, Oita, 879–5593 Japan (H. Yoshimatsu,
K. Tsuda, M. Tatsukawa, S. Chiba, T. Sakata); Department of
Physiology, Niigata University, School of Medicine, Niigata,
951 Japan (A. Niijima).
Correspondence to: Toshiie Sakata, MD, PhD, Professor,
Department of Internal Medicine I, School of Medicine, Oita
Medical University, 1–1Idaigaoka, Hasama, Oita, 879–5593 Japan.
Tel.: + 81–97–586 –5790; fax: + 81–97–549–4480; e-mail:
sakata@oita-med.ac.jp
Received 30 July 2001; accepted 26 November 2001
shown to suppress food intake through H1 receptors in the
ventromedial hypothalamic and the paraventricular nuclei
[1,2]. Central administration of leptin increased histamine
turnover rate [3], which was assessed by accumulation of
tele-methylhistamine, a major metabolite of neuronal his-
tamine. Leptin-induced suppression of feeding was attenu-
ated in the rats [3] in which neuronal histamine was
depleted by the administration of α-fluoromethylhistidine
(FMH), a suicide inhibitor of histamine-synthesizing
enzyme, histidine decarboxylase. Hypothalamic neuronal
histamine was thus identified as one of the targets of leptin
action in the brain.

© 2002 Blackwell Science Ltd


Recently, hypothalamic histamine neurons were found to
modulate peripheral lipid metabolism as well as energy
expenditure. Activation of hypothalamic histamine neurons
increased lipolysis in white adipose tissue (WAT) through
activation of sympathetic nerve [4], and up-regulated
mRNA expression of uncoupling protein 1 in brown adipose
tissues [5]. Thus, hypothalamic histamine neurons play a
crucial role in the maintenance of energy homeostasis through
central regulation of adiposity not only by affecting food
intake, but also by affecting peripheral energy expenditure.
The semiessential amino acid, L-histidine, is a precursor
of neuronal histamine. Peripheral administration of his-
tidine increased histamine concentration in the hypo-
thalamus [6– 8]. However, there have been few studies to
demonstrate that peripheral loading of histidine affects
physiological brain functions. Our recent study showed that
peripheral administration of histidine induced suppression
of feeding, which was attenuated by the depletion of
neuronal histamine using FMH [8]. Our data suggest that
following histidine loading, the histidine-induced suppres-
sion of feeding is preceded by the conversion of histidine
to histamine in the hypothalamus. The present study aimed
to investigate the involvement of peripheral histidine load
on the control of lipolysis through sympathetic nerve
activation. The physiological implications of histidine are
discussed as an activator of histamine neurons in the
hypothalamus.
Materials and methods
Animals and diet
Mature male Wistar King A rats weighing 280 –320 g were
used. They were housed in a room illuminated daily from
08·00 h to 20·00 h (a 12 : 12 h light–dark cycle) and main-
tained at 21 ± 1 °C with humidity at 55 ± 5%. All rats were
allowed free access to pelleted rodent chow (#CE-2; Clea
Japan Ltd, Tokyo, Japan) and tap water. All studies were
conducted in accordance with the Oita Medical University
Guidelines that are based on the National Institute of
Health Guide for the Care and Use of Laboratory Animals.
Reagents
Isoproterenol, a β-adrenoceptor agonist, L-histidine and L-
histamine (Sigma, St. Louis, MO, USA) were dissolved in
phosphate buffered saline (PBS) to concentrations of 10–8
to 10–6 M, 10–3 M and 10–2 M, respectively. Each solution
was freshly prepared on the day of its administration. The
pH of each solution was adjusted to a range from 6·4 to 7·2.
Surgery
Rats that were to be used for central infusions underwent
surgery for placement of indwelling cannula. Under sodium
© 2002 Blackwell Science Ltd, European Journal of Clinical Investigation, 32, 236–241
Histidine increases lipolysis 237
pentobarbital anesthesia (45 mg kg–1, ip), the rats were fixed
in a stereotaxic apparatus (Narishige Co., Tokyo, Japan)
and a 23G-stainless steel guide cannula was chronically
implanted into the third cerebroventricle (i3vt). A 29G-
stainless steel wire stylet was left in the guide cannula to
prevent the leakage of cerebrospinal fluid and obstruction
of the cannula. Rats were allowed to recover from surgery
for at least 1 week before any treatments were administered.
Rats that were to be used for blood collection underwent
surgery under ether anaesthesia for implantation of chron-
ically indwelling silicone catheter in the right external
jugular vein with its end at a point just outside of the atrium
[9]. Surgical catheterizations were performed 7 days prior
to the histidine injections.
For in vivo microdialysis study, a small incision was made
under sterile conditions on the skin in the inguinal area. A
probe was inserted and fixed in the epididymal WAT
through the incision with the aid of a guide cannula. Details
of the surgical procedures have been described elsewhere
[4,9].
In vivo microdialysis to assess lipolytic activity
Microdialysis study was performed on anaesthetized rats
(pentobarbital 45 mg kg–1) [4,10,11]. A dialysis tube
(0·5 mm long dialysis membrane with a molecular cut-off
of 50 kDa) surrounding a double-lumen microinjection
stainless steel cannula (Eicom Co Ltd, Tokyo, Japan) was
used. The perfusion solution entered the probe through the
inner cannula and passed down to the tip of the probe.
Thereafter, it streamed upwards in the space between the
inner cannula and the outer dialysis membrane. The dialy-
sate left the probe through the outer cannula via a sidearm
from which it was collected. During dialysis, the probe was
connected to a microinjection pump (Eicom Co. Ltd,
Tokyo, Japan).
The microdialysis probe was allowed to equilibrate
40 min following its implantation. Seven rats were used to
study the effect of isoproterenol on adipose tissue lipolysis.
In vivo microdialysis study was started by perfusing PBS
through the perfusion cannula at a rate of 5 µL min–1 for
60 min After this, PBS (control treatment) and isoprotere-
nol at concentrations of 10– 8 to 10–6 M were sequentially
administered through the perfusion cannula at a rate of
5 µL min–1 for 120 min (30 min for each solution). The dia-
lysate samples were collected over 15 minute period from
the outlet of the polyethylene probe, and each aliquot
was assayed for glycerol. Glycerol was assayed using
20 µL sample of each dialysate aliquot collected, using an
automated luminescence analyzer (Lumat # LB9501,
Berthold Wildbat, Waildbad, German) [12,13]. Test
samples were assayed immediately after the calibration
of the microdialysis probes with standards on the same
experimental day.
A second group of rats was used to study the effect of
peripheral administration of histidine on adipose tissue
lipolysis. Following equilibration of the microdialysis probe,
in vivo microdialysis was started by perfusing PBS through
238 H. Yoshimatsu et al.

the perfusion cannula at a rate of 5 µL min–1. Histidine was

administered at a dose of 0·35 mmol kg–1 ip, or PBS (n =
5 per group). Dialysate samples were collected 30 min after
histidine administration.
A third group of rats (previously implanted with i3vt
cannula) was used to study the effect of i3vt infusion of
histamine on adipose tissue lipolysis. Following equilibration
of the microdialysis probe, in vivo microdialysis was started
by perfusing PBS through the perfusion cannula at a rate
of 5 µL min–1. Histamine at a dose of 100 nmol rat–1, or
PBS was infused through the i3vt cannula at a rate of
1 µL min–1 for 10 min (n = 7 per group). Dialysate samples
were collected 30 min after histamine administration to
measure glycerol release.

Blood sampling and assay of free fatty acid

Rats previously implanted with chronically indwelling

jugular vein catheters were used. Rats were administered
histidine at a dose of 0·35 mmol kg–1 ip, or PBS, and blood
samples were collected from the jugular vein with catheters.
Serum samples were immediately frozen at –20 °C until
assay on the following day for humoral factors. The
concentration of free fatty acid (FFA) in the serum was
measured using a commercially available kit (NEFA-SS’
Eiken’; Eiken Chemical Co. Ltd, Tokyo, Japan).
Figure 1 Effects of various concentration of isoproterenol (10– 8 to
10– 6 M) in the perfusate on glycerol concentration in the dialysate
from epididymal adipose tissue. Values and vertical bars,
means ± SEM (n = 7). *P < 0·05 vs. phosphate buffered saline
(PBS) controls.
Statistical analysis

All values were expressed as means ± SEM. Data was

Recording of sympathetic nerve activity
Electrophysiological recordings were carried out under
urethane anaesthesia (1·0 g kg–1). The left epididymal adipose
tissue was exposed through a left inguinal incision. Using
a dissecting microscope, nerve filaments were isolated from
the sympathetic nerves innervating the left epididymal
adipose tissue. To differentiate neuron activity of efferent
nerves from that of afferent nerves, the peripheral connec-
tion of the recording nerve to the epididymis was cut.
Efferent discharges were recorded from fine filaments of
sympathetic nerve fibers that were dissected free from con-
nective tissue nerve activity was detected by a pair of silver
wire electrodes that were immersed in a mixture of liquid
paraffin and white Vaseline to prevent dehydration. The
action potential was amplified by means of a conventional
differential amplifier. All analyses of nerve activity were
based on the records obtained after the conversion of raw
data to standard pulses using a window discriminator
that distinguishes nerve activity from background noise.
Impulses were integrated by a rate meter with a reset time
of 5 s and recorded by a pen recorder. After determining
the background firing rate of sympathetic nerves, the effect
of ip histidine (0·35 mmol kg–1) and PBS administration on
the nerve activity was investigated (n = 5 for each group)
over a 120 min maximum recording time. Each measure-
ment was made at –15 min, 0 (immediately before), and 30,
60, 90 and 120 min after the histidine injection. Details
of the recording of nerve activity have been described
elsewhere [4,14,15].
analysed using Student’s t-test and two-way Analysis of
Variance (ANOVA) with a replication method followed by
Scheffe’s post hoc test. Evaluation of a dose-responsiveness
was carried out by a single linear regression and ANOVA.
Results
Effect of isoproterenol on lipolysis
Figure 1 shows the changes of glycerol concentration in the
dialysate of epididymal adipose tissue following sequential
perfusion of PBS, and isoproterenol at concentrations rang-
ing from 10–8 to 10–6 M. At steady state conditions following
PBS perfusion, the mean glycerol concentration in adipose
tissue dialysate was 18·1 ± 3·4 µM, reflecting basal lipolysis.
When the concentration of isoproterenol in the perfusate
was gradually and sequentially increased from 10– 8 to 10– 6 M,
there was a significant elevation of glycerol concentrations
compared to the basal level (PBS perfusion) (P < 0·05).
This increase in glycerol release was dose dependant
( Y = 33·267 + 0·001X, r = 0·826, P < 0·01; Y, glycerol
concentration; X, logarithm of isoproterenol dosage).
Effects of L-histidine and histamine on lipolysis
Figure 2 shows changes of glycerol concentration in the
dialysate from epididymal adipose tissue 30 min after ip

© 2002 Blackwell Science Ltd, European Journal of Clinical Investigation, 32, 236 – 241

Figure 2 Effects of ip injection of L-histidine (0·35 mmol kg–1) or
i3vt infusion of histamine (100 nmol rat–1) on glycerol
concentration in the dialysate of epididymal adipose tissue. Values
and vertical bars are means ± SEM for histidine (n = 5), and
histamine (n = 7). *P < 0·05 vs. PBS controls.
injection of 0·35 mmol kg–1 histidine, or i3vt infusion of
histamine at a dose of 100 nmol rat–1. Administration of
histidine significantly increased glycerol concentrations
in the dialysate compared with PBS controls (P < 0·05).
Similar to the results with histidine administration, i3vt
infusion of histamine increased glycerol concentration
more potently than PBS infusion (P < 0·05). Peripheral
administration of histidine also increased serum level of
FFA compared to PBS controls (P < 0·05) (Figure 3).
Effect of histidine on WAT sympathetic activity
Changes in activity of the sympathetic efferent nerves
innervating the left epididymal adipose tissue following ip
injection of histidine are shown in Fig. 4 as percent change
from the baseline firing rate. The mean discharge rate at
–15 min, 0 (immediately before), 30, 60, 90 and 120 min
after the administration of histidine was 85·5 ± 8·3,
83·8 ± 5·1, 113·1 ± 7·2, 125·7 ± 12·0, 111·5 ± 11·7 and
104·8 ± 10·5 impulses/5 s, respectively. PBS infusion did
not result in any remarkable change in sympathetic
nerve activity. There was a significant difference in response
of the sympathetic nerve activity to histidine administra-
tion compared to that following PBS administration
(P < 0·05).
Discussion
The present study has demonstrated that peripheral admin-
istration of -histidine accelerated lipolysis in WAT. Follow-
ing the release of lipolytic substance such as catecholamine
secreted by the adrenal medulla, and glucagon from the
pancreas, the triglycerides deposited in adipose tissue are
© 2002 Blackwell Science Ltd, European Journal of Clinical Investigation, 32, 236–241
Histidine increases lipolysis 239
Figure 3 Effects of ip injection of histidine (0·35 mmol kg–1) on
serum level of free fatty acid. Values and vertical bars are
means ± SEM (n = 5). *P < 0·05 vs. PBS controls.
metabolized into glycerol and fatty acids. Recently, WAT has
been shown to receive direct sympathetic innervation from
the spinal cord [16] and origins of this efferent pathway in
the central nervous system have been identified in several
nuclei of the brain stem and the hypothalamus [16]. Func-
tionally, cold exposure has been shown to increase turnover
rate of norepinephrine in WAT together with circulating
concentration of FFA [17]. These findings indicate the
involvement of increased sympathetic nerve activity in lipo-
lytic process of adipocytes. Consistently with those previous
findings, the present study proved that direct infusion of
isoproterenol, a β adrenoceptor stimulant, into adipose
tissue increased glycerol concentration in the dialysate in a
dose-dependent manner.
In the present study, intraperitoneal administration of
histidine resulted in an increase in the glycerol concentration
in the dialysate from epididymal fat pad, as well as an
elevation in serum FFA concentration, indicating lipolysis
in progress. Intraperitoneal administration of histidine
mimicked the enhancement of lipolysis induced by i3vt
infusion of histamine. Here, it can be debated as to whether
histamine concentration in the hypothalamus following
peripheral administration of histidine may be compatible with
that after icv histamine infusion. There is no definite answer
to the question to date. One possible explanation based on
our present and previous studies may explain the query
convincingly. According to our previous study in which
histidine was injected ip at the same dose and in the same
methodological procedures with those in the present study,
there was 2 nmol g–1 tissue weight difference between whole
hypothalamic histamine levels before and after histidine
injection [8]. This increased value was based on the histamine
240 H. Yoshimatsu et al.
level in dissected tissue weight of the hypothalamus
(100 mg) but not in discrete hypothalamic nuclei. Evidence
shows that exogenous administration of histidine dose not
diffusely elevate histamine concentration in the hypothala-
mus, because the distribution of histidine decarboxylase,
histamine synthesizing enzyme, is known to localize heavily
in these discrete nuclei [18]. Histamine elevation in the
hypothalamus thus implicates enhancement of neuronal
histamine localized in some specific nuclei, i.e. the tubero-
mammillary nucleus where histamine cell bodies are
localized, the ventromedial hypothalamic nucleus, the
paraventricular nucleus (histamine neurons project densely
to the latter two nuclei) and so on. Elevation of net histamine
concentration in those discrete nuclei is expected to be
much higher than the measure detected in the whole
hypothalamus (at least more than 10 times higher than a
measure of 2·0 nmol g–1). In other words, it is quite probable
that net histamine in discrete region reaches 20 nmol g–1
tissue weight following histidine treatment. Conversely, icv
infusion study has demonstrated that it is possible to get
exogenously infused neurotransmitters into the hypotha-
lamic nuclei merely at a ratio of 1/50~1/100 of the total
infusion dose because of diffusion into the cerebrospinal
fluid [19]. Adjusting to the ratio, the amount of histamine
diffused into the hypothalamus in the present icv study
is assessed at 10–20 nmol g–1 tissue weight. According to
those calculations, there may be a negligible difference
in hypothalamic histamine concentration between the
procedures with ip histidine and icv histamine. Indeed,
the assessment appears reasonable because the results of
lipolytic activity were consistent in both the present ip
histidine and icv histamine injection studies.
In our previous study using high performance liquid chro-
matography, both histamine concentration and activity of
histidine decarboxylase in the hypothalamus were increased
after the ip administration of histidine at the same dose as
that used in the present study [8]. These findings indicate
the involvement of histidine in adipose tissue lipolysis
through activation of hypothalamic histamine neurons,
although a direct effect of peripheral histidine on lipolysis
cannot be excluded. Intracerebroventricular infusion of
histamine has been reported to increase blood concentration
of catecholamine [20]. A previous study showed that i3vt
infusion of thioperamide, a histamine H3 receptor antago-
nist that accelerates synthesis and release of neuronal
histamine at the presynaptic nerve terminal, increased
activity of efferent sympathetic nerve innervating adipose
tissue [4]. In this in vivo microdialysis study, pretreatment
with propranolol, a β-adrenoceptor antagonist, completely
blocked thioperamide-induced lipolysis [4].
The present results indicate that activation of hypotha-
lamic histamine neurons leads to sympathetic augmentation
of lipolytic activity in adipose tissue. To confirm the involve-
ment of an efferent pathway from hypothalamic histamine
neurons to adipose tissue in histidine-induced lipolysis, we
investigated the effects of histidine on sympathetic nerve
activity using an electrophysiological method. The results
showed that peripheral administration of histidine increased
activity of sympathetic nerve innervating WAT.
© 2002 Blackwell Science Ltd, European Journal of Clinical Investigation, 32, 236 – 241
Figure 4 The effect of ip injection of histidine on activity of efferent
sympathetic nerve innervating left epididymal adipose tissue.
Results are represented as the percentage difference in sympathetic
nerve activity from baseline value (100%) after ip injection of
0·35 mmol kg–1 histidine. The arrow indicates injection of test
solutions. Values and vertical bars are means ± SEM (n = 5).
*
P < 0·001 vs. PBS controls.
Recent studies have shown a novel role of histamine
neurons as a target of leptin action in the hypothalamus.
Hypothalamic histamine neurons were activated by central
infusion of leptin [3]. Leptin-induced suppression of feed-
ing was attenuated in rats in which the histamine had been
depleted with FMH, and in histamine H1 receptor knock-
out mice [3,21]. The results suggest that feeding behavior
regulated by leptin is modulated by neuronal histamine
through H1 receptor. In addition, leptin-induced up-
regulation of uncoupling protein 1 mRNA in brown adipose
tissue was attenuated in histamine H1 receptor knockout
mice [21]. Signal transduction from leptin to histamine
neurons plays an essential role in regulation of energy
metabolism. In fact, disruption of this signaling has been
detected in genetic obese animal models, such as obese
ob/ob and diabetic-obese db/db mice, and fatty fa/fa rats in
which hypothalamic histamine concentrations are lowered
because of defects in leptin signal [3,22,23].
It is generally accepted that leptin regulates adiposity
through its hypothalamic inhibitory effects on food intake
and the stimulatory effects on energy expenditure. How-
ever, in most obese animals and humans, exogenous admin-
istration of leptin is ineffective on weight reduction because
of their leptin resistance [24 –26]. It is greatly expected that
manipulation of neuromodulators or their receptors that
exist in downstream pathway of leptin, but not leptin per se,
may be useful targets for developing antiobesity agents.
Among such neuromodulators, the activation of hypotha-
lamic histamine neurons by peripheral loading of histidine
is considered to be the most physiological treatment because
foodstuffs normally provide the semiessential amino acid,
histidine. Taken together, an oral load of histidine, even in
the presence of leptin resistance, may lead to activation of

neuronal histamine that stimulates lipolysis in adipose
tissues through sympathetic innervation.
Acknowledgements
We thank Dr Narinder. S. Shargill, Fremont, CA, USA, for
help with the manuscript. Supported partly by Grants-
in-Aid 02454129 (TS) from the Japanese Ministry of
Education, Science and Culture; by Research Grants for
Intractable Diseases from the Japanese Ministry of Health
and Welfare, 2000.
References
1 Sakata T, Ookuma K, Fukagawa K, Fujimoto K,
Yoshimatsu H, Shiraishi T et al. Blockade of the histamine
H1-receptor in the rat ventromedial hypothalamus and feeding
elicitation. Brain Res 1989;441:403–7.
2 Sakata T, Yoshimatsu H. Hypothalamic neuronal histamine:
Implications of homeostatic maintenance in its control of
energy metabolism. Nutrition 1997;13:403–11.
3 Yoshimatsu H, Itateyama E, Kondou S, Tajima D, Himeno K,
Hidaka S et al. Hypothalamic neuronal histamine as a target of
leptin in feeding behavior. Diabetes 1999;48:2286–91.
4 Tsuda K, Yoshimatsu H, Niijima A, Hidaka S, Okeda T,
Sakata T. Hypothalamic histamine neurons activate lipolysis
in rat adipose tissue. Exp Biol Medical 2002;227:208–10.
5 Masaki T, Yoshimatsu H, Chiba S, Watanabe T, Sakata T.
Central infusion of histamine reduces fat accumulation and
upregulates UCP family in leptin-resistant obese mice. Diabetes
2001;50:376 – 84.
6 Schwartz JC, Lampart C, Rose C. Histamine formation in rat
brain in vivo: effects of histidine loads. J Neurochem
1972;19:801–10.
7 Taylor KM, Snyder SH. Dynamics of the regulation of
histamine levels in mouse brain. J Neurochem 1972;19:341–54.
8 Yoshimatsu H, Chiba S, Tajima D, Akehi Y, Sakata T. Histidine
suppresses food intake through its conversion into neuronal
histamine. Exp Biol Medical 2002;227:63 – 8.
9 Sakata T, Tsutsui K, Fukushima M, Arase K, Kita H,
Oomura Y. Feeding and hyperglycemia induced by 1·5-
anhydroglucitol in the rat. Physiol Behav 1981;27:401–5.
10 Tossman U, Ungerstedt U, Arner P. Microdialysis in the study
extracellular levels of amino acids in the rat brain. Acta Physiol
Scand 1986;128:9 –14.
11 Arner P, Bolinder J, Eliasson A, Lundin A, Ungerstedt U.
Microdialysis of adipose tissue and blood for in vivo lipolysis
studies. Am J Physiol 1988;255:E737–42.
© 2002 Blackwell Science Ltd, European Journal of Clinical Investigation, 32, 236–241
Histidine increases lipolysis 241
12 Bjorkhem I, Arner P, Thore A, Ostman J. Sensitive kinetic
bioluminescent assay of glycerol release from human fat cells.
J Lipid Res 1981;22:1142–7.
13 Hellmer J, Arner P, Lundin A. Automatic luminometric kinetic
assay of glycerol for lipolysis studies. Anal Biochem
1989;177:132–7.
14 Niijima A. The effect of 2-deoxy--glucose and -glucose on
the efferent discharge rate of sympathetic nerves. J Physiol
1975;251:231–43.
15 Yoshimatsu H, Niijima A, Oomura Y, Yamabe K, Katafuchi T.
Effects of hypothalamic lesion on pancreatic autonomic nerve
activity in the rat. Brain Res 1984;303:147–52.
16 Bamshad M, Aoki VT, Adkison MG, Warren WS, Bartness TJ.
Central nervous system origins of the sympathetic nervous
system outflow to white adipose tissue. Am J Physiol
1998;275:R291–R299.
17 Garofalo MAR, Kettelhut IC, Roselino JES, Migliorini RH.
Effect of acute cold exposure on norepinephrine turnover
rates in rat white adipose tissue. J Auton Nerv Syst
1996;60:206–8.
18 Scwartz JC, Arrang KM, Garbarg M, Pollard H, Ruat M.
Histaminergic transmission in the mammalian brain. Physiol
Rev 1991;71:1–51.
19 Myers RD, Tytell M, Kawa A, Rudy T. Microinjection of 3H-
acetylcholine, 14C-serotonin and 3H-norepinephrine into the
hypothalamus of the rat: diffusion into tissue and ventricles.
Physio Behav 1971;7:743–51.
20 Knigge U, Matzen S, Warberg J. Histamine as a neuroendocrine
regulator of the stress induced release of peripheral
catecholamines. Endocrinology 1990;126:1430 – 4.
21 Masaki T, Yoshimatsu H, Chiba S, Watanabe T, Sakata T.
Targeted disruption of histamine H1 receptor attenuates
regulatory effects of leptin on feeding, adiposity, and UCP
family in mice. Diabetes 2001;50:385 –91.
22 Machidori H, Sakata T, Yoshimatsu H, Ookuma K,
Fujimoto K, Kurokawa M et al. Zucker obese rats:
defect in brain histamine control of feeding. Brain Res
1992;590:180–6.
23 Yoshimatsu H, Machidori H, Doi T, Kurokawa M,
Ookuma K, Kang M et al. Abnormalities in obese Zuckers:
Defective control of histaminergic functions. Physiol Behav
1993;54:487–91.
24 Maffei M, Halaas J, Ravussin E, Pratley RE, Lee GH,
Zhang Y et al. Leptin levels in human and rodent: measurement
of plasma leptin and ob RNA in obese and weight-reduced
subjects. Nature Med 1995;1:1155–61.
25 Concidine RV, Sinha MK, Heiman ML, Kuriauciunas A,
Stephens TW, Nyce M et al. Serum reactive-leptin
concentrations in normal weight and obese humans. N Engl J
Med 1996;334:292–5.
26 van Heek M, Compton DS, France CF, Tedesco RP,
Fawzi AB, Graziano MP et al. Diet-induced obese mice develop
peripheral, but not central, resistance to leptin. J Clin Invest
1997;99:385–90.