Presented by Int

2013/4/25
Introduction
males display greater degree of antinociception
than females
-- Cicero et al., 1996; Boyer et al., 1998; Kest et al., 1999; Craft and Bernal, 2001.
Male rats develop greater tolerance to
morphine than females
--Badinllo-martines et al., 1984;Craft et al., 1999
sex steroids mediate these differences by acting
on neuronal processes in CNS
--Cicero et al., 2002; Craft et al., 2004
No study to report the mechanisms of sex-related
differences in tolerance to morphine
known role of excitatory amino acids including
glutamate and it's receptors in the development of
tolerance to opioids
--Fundytus and Coderre, 1994; Mao et al., 1994
There is evidence on the potentiating actions of E2 on
glutamate-induced excitation --Smith, 1989

it is supposed that gonadal steroids are responsible for
sex-related differences in morphine tolerance via
effecting glutamate concentration in CNS.
Introduction
nucleus accumbens is critically involved in
the development of tolerance to opioids
--Gracy et al., 1997; Schmidt et al., 2002
there are functional associations between
glutamate receptors and morphine in this
brain region --Gracy et al., 1997

glutamate level changes in nucleus
accumbens may bear a relationship with sex
differences in morphine tolerance.
Introduction
Materials and methods
Male
Control(Saline)
Intact
GDX
Sham
(open close)
female
Control(Saline)
Intact
GDX
Sham
(open close)
Gonadectomy:
Females: ovariectomized by removing the
ovaries and ovarian fat via dorsal incisions.
Males: castrated by removing the testes
and testicular fat following 2 cm midscrotal
incision.
Sprague-Dawley rats,
221 C, 12 h light/dark cycle
Morphine sulfate (7 mg/kg, s.c.)
x 8 consecutive days
10s of heat to prevent tissue injury
D1 D2 D3 D4 D5 D6 D7 D8
20mins
test
D1/4/6/8
application of thermal radiation to the tail of an
animal provokes the withdrawal of the tail by a brief
vigorous movement
--D'Amour and Smith, 1941;Smith et al., 1943
Baseline
Materials and methods
Tail flick time test
Materials and methods
D1 D2 D3 D4 D5 D6 D7 D8
ACSF: NaCl 114 mM, KCl 3 mM, CaCl 2 1 mM, MgSo 4 2 mM, NaH 2 PO 4 1.25 mM,
NaHCO 3 26 mM, NaOH 1 mM, glucose 10 mM and pH=7.4
Baseline

D1
Artificial CSF pump,3.0 l/min
D8
sample
30mins
20mins
Glutamate was measured as its
O-phthalaldehyde (OPA)/beta-mercaptoethanol
derivative using a high-performance liquid
chromatography with fluorescence detector system
Glutamate level test
Tolerance was defined as a decrease in, or the
loss of the antinociceptive effect of a single
dose of morphine on first day.
The latency differences between the first and
8th day of experiment were considered as the
degree of tolerance.
all data were analyzed by one-way analysis
of variance (ANOVA) followed by a series of
priori comparison contrasts to evaluate
interactions of interest.

Materials and methods
Results
Influences of sex and gonadal hormones in the
development of tolerance to morphine antinociception
Tail flick time test
Results
0
1
2
3
4
5
6
7
8
9
10
baseline D1 D4 D6 D8
M, Intact
M, GDX
F, Intact
F, GDX
Degree of tolerance
Tail flick time test
Results
Effects of sex and gonadal hormones on nucleus accumbens
glutamate level in morphine tolerant rats
insignificant
Glutamate level test
males showed greater morphine
tolerance than females
-similar results in only one previous report
(Craft et al., 1999)

Discussion
Discussion
acute dose of morphine has typically greater analgesic
activity in male rats than in females.
morphine was a more potent analgesic in male than in female
rodents --Mogil et al., 2000; Craft, 2003; Holtman et al.,2004

BUT magnitude of sex differences in the present study may
differ from that reported by some investigators
Why different?
strain of rodents
--Kasson and George, 1984; Kest et al., 1999, 2000a
applied doses of morphine or analgesic testing models
--Bartok and Craft, 1997; Kest et al., 2000b
vendors from which animals are purchased
--Craft et al., 1999
Gonadectomy of animals decreased the magnitude of
sex differences to non-significant levels
confirms the role of gonadal hormones in sex-related
differences in morphine analgesia
--Craft et al., 2004; Fillingim and Gear, 2004
Our results provide the first evidence for the role of gonadal
hormones on sex differences in morphine tolerance
development.
sex differences in morphine tolerance disappeared in gonadectomized
animals
but further examinations are needed to determine the role of each
kind of gonadal hormones in these effects.

Discussion
Discussion
Significant differences of nucleus accumbens glutamate
level in gonadally intact female rats versus males
the involvement of excitatory amino acid neurotransmission in
sex related differences in morphine's antinociception effect.

involvement of glutamate mediated synaptic transmission in
the opiate abuse and tolerance
--Hakan and Henriksen, 1989
Particularly, NMDA receptors
--Hsu and Wong, 2000; Wen et al., 2004

There is decrease in extracellular level of glutamate in brain by
morphine
--Hao et al., 2005

relationship between excitatory amino acid release and lack of
an antinociceptive response to morphine
--Wen et al., 2004
Discussion
Glutamate concentration shows
approximately 30% decrease in tolerant male rats
versus 200% increase in females
This may explain the rapid development of tolerance in
females than in males
increasing CSF excitatory amino acids concentration has been
shown in morphine tolerant rats
--Akaola and Aston-Jones, 1991; Jhamandas et al., 1996; Wen et al., 2004
Chronic administration of morphine may increase the synaptic
excitatory amino acids levels and activate glutamate receptors,
which may result in reduction of the nociceptive threshold
and the morphine antinociception
--Wen et al., 2004

The presence of estrogen and progestin receptors in
regions involved in pain sensitivity (e.g. central gray)
has also been demonstrated
--McEwen and Alves, 1999
ovarian hormones interact with NMDA receptors to
induce new synapses.
Confocal microscopic imaging has shown that
estrogen treatment upregulates immunoreactivity
for the largest NMDA receptor subunit and cell
bodies of some neurons in brain
--McEwen, 2001
Discussion
Therefore, high level of estrogens in females may be
responsible for the noticeable increase in the nucleus
accumbens glutamate concentration in this study.
Estrogen induction of new excitatory synapses in
pain pathways may also explain the lower morphine
antinociception in female rats than in males.
Further investigations are required to clarify the
exact molecular mechanisms or interactions between
gonadal steroids and pain modulatory systems.
Conclusion
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