Sleep analysis in mice laking lipocalin-type prostaglandin D synthase

s221

458

STUDIES ON THE SLEEP-PROMOTING

MECHANISMS

MEDIATEDBY

ADENOSINEA,,

RECEPTORS.

Satoh, S., Koike, N., and Hayaishi, 0. Dept. of Molecular Behavioral Biology, Osaka Bioscience Institute, Osaka 565-0874, Japan We previously reported that an adenosine hA receptor agonist, CGS21680, produced profound increases in non-rapid-eyemovement (NREM) sleep and rapid-eye-movement (REM) sleep in rats when this compound was administered to the prostaglandin D,-sensitive, sleep-promoting zone, namely the subarachnoid space underlying the rostral basal forebrain. In this paper, we proposed the site of action of CGS21680 for sleep-promotion and confirmed the receptor subtypes involved in the sleep-promotion in this region. Firstly, we administered CGS21680 to various parts of bram and compared the respective effects on sleep. Then to visualize the neuronal elements activated by CGS21680, we examined the expression of Fos protein following the administration of CGS2 1680 to the most effective area for sleep-promotion. Furthermore, to clarify the receptor subtypes involved in the sleep promotion in this brain region, we compared the sleep-promoting effects of five adenosine receptor agonists; namely CGS21680, APEC, NECA, CHA, and CPA. The results indicated that (1) The largest increases m NREM sleep and REM sleep appeared when CGS21680 was administered to the subarachnoid space underlying the rostra1 basal forebrain. (2) Populations of Fos immunoreactive neurons appeared within the medial part of the ventral striatum where an intense distribution of adenosine hZA receptor are known. (3) CGS21680, APEC, and NECA produced the increases m NREM sleep and REM sleep at the administration rate of 2.0 pmolimin, but the same dose of CPA and Cl-IA did not promote either types of sleep.These findings were interpreted to mean that the activities of adenosine A,, receptors localized in ventral striatum, if not only, participate in the promotion of sleep. THE SOMNOGEN PGD2 INDUCES SLEEP-ACTIVE NEURONS

459

DMITRY GERASHCHENKO’; SAPER4, OSAMU HAYAISHI’.

THOMAS

FOS EXPRESSION

SCAMMELL’;

WITHIN UNIQIJE POPULATIONS

HIROTAKA

ONOE’; YOSHIHIRO

OF

URADE’,+

CLIFFORD

‘Dept. of Molecular Behavioral Biology, and 2Dept. of Neuroscience, OBI, Furuedai 6-2-4, Suita, Osaka 5650874; ‘CREST, Japan Science and Technology Corp.; Dept. of Neurology, “Harvard Medical School, Boston, MA 02215, USA Prostaglandin

D2 (PGD2) induces sleep specifically

and dose-dependently

when it is infused into the ventral surface of the

rostra1 basal forebrain in rats at very low concentrations

(I-200

pmol/min).

Because

activated neurons, we applied FOS immunohistochemistry

to study the mechanisms

FOS protein accumulates

in recently

of sleep induction by PGD2. Compared

to the prior baseline day, infusion of PGD2 at the rate of 200 pmoUmin highly increased

amount of slow wave sleep (SWS)

during second hour of infusion (21:00-22:OO; SWS increased to 29.9 min from baseline amount of 7.7 min). PGD2-treatment markedly increased number of FOS-immunoreactive, In contrast, expression

it almost completely was clearly observed

suppressed

FOS expression

preoptic area (VLPO) (p < 0.00 I).

in the tuberomammillary

in this area in control awakin g rats infused

strongly support the crucial role of the VLPO-TMN mechanism

activated neurons in the ventrolateral

nucleus

(TMN),

with saline (1, < 0.002).

neuronal network in sleep regulation

whereas

FOS

Our observations

and provide a new insight into the

of sleep induction by PGD2.

SLEEP ANALYSIS

460

IN MICE LAKING LIPOCALIN-TYPE

NAOMI EGUCHI’,2; YUKO KUWAHATA’; YOSHIHIDE YOSHIHIRO URADE=; OSAMU HAYAISHF.

KANAOKA’;

PROSTAGLANDIN AKIHISA

D SYNTHASE

NAGATA’,

NOBUAKI

YOSHIDA’,

‘Dept. Morphol. Brain Sci., Fat. Med., Kyoto Univ., Kyoto 606-8315; ‘CREST (JST), Osaka 5650874; ‘Fat. Med., Univ. Tokyo; 4Res. Inst. Osaka Med. Centr. Maternal Child Health, Osaka 594-l 101; and ‘Osaka Biosci. Inst., Osaka 565-0874 Prostaglandin

(PG) D2 is a major prostanoid

potent endogenous

sleep-inducing

substance.

in the CNS of various mammals Lipocalin-type

including

PGD synthase (L-PGDS)

in the CNS and localized in the leptomeninges,

choroid plexus, and oligodendrocytes.

kb) containing

from a 129/Sv genomic

homologous

the gene for mouse L-PGDS

recombination

three independent deficiency

abnormalities

gene on sleep, we monitored

gene does not lead to fetal death. as judged by visual observation.

L-PGD.Y’To investigate

sleep of L-PGDS-‘- mice by measuring

implantable telemetric transmitter we have developed.

and known

is responsible

as the most

for PGD2 biosynthesis

We isolated the genomic clone (15.3

library and constructed gene.

a targeting

vector used for

The chimeric male mice were produced

mutated ES clones and mated with female mice of an inbred

of the L-PGDS

morphological

to produce the null mutation of the L-PGDS

humans,

129 strain to generate

mice had normal

homozygotes.

behavior

from The

and no apparent

the effect of the null mutation of the L-PGDS

the electroencepharogrum

and electromyogram

with an