Estradiol-Induced Potentiation of Dopamine Release in Dorsal Striatum Following Amphetamine Administration Requires Estradiol Receptors and mGlu5.

Estradiol potentiates behavioral sensitization to cocaine as well as self-administration of cocaine and other drugs of abuse in female rodents. Furthermore, stimulated dopamine (DA) in the dorsolateral striatum (DLS) is rapidly enhanced by estradiol, and it is hypothesized that this enhanced DA release mediates the more rapid escalation of drug taking seen in females, compared with males. The mechanisms mediating the effect of estradiol to enhance stimulated DA release were investigated in this study. Using in vivo microdialysis and high performance liquid chromatography coupled with electrochemical detection, we first examined the effect of estradiol on amphetamine-induced DA increase in the DLS of ovariectomized rats. We then tested whether the potentiation of this DA increase could be blocked by the estradiol receptor antagonist, ICI 182,780 (ICI), or an antagonist to the metabotropic glutamate receptor subtype 5 (mGlu5), 2-methyl-6-(phenylethynyl)pyridine (MPEP). There is evidence that estradiol receptors collaborate with mGlu5 within caveoli in DLS and mGlu5 is hypothesized to mediate many of the effects of estradiol in the addiction processes in females. Our data show that estradiol enhances the DA response to amphetamine. Either ICI or MPEP prevented the effect of estradiol to enhance DA release. Importantly, our results also showed that neither ICI or MPEP alone is able to influence the DA response to amphetamine when estradiol is not administrated, suggesting that ICI and MPEP act via estradiol receptors. Together, our findings demonstrate that estradiol potentiates amphetamine-stimulated DA release in the DLS and this effect requires both estradiol receptors and mGlu5.

Estradiol acts during a post-pubertal sensitive period to shorten free-running circadian period in male Octodon degus.

The free-running circadian period is approximately 30 min shorter in adult male than in adult female Octodon degus. The sex difference emerges after puberty, resulting from a shortened free-running circadian period in males. Castration before puberty prevents the emergence of the sex difference, but it is not a function of circulating gonadal hormones as such, because castration later in life does not affect free-running circadian period. The aim of this study was to determine whether or not the shortening of the free-running circadian period in male degus results from exposure to gonadal hormones after puberty. We hypothesized that masculinization of the circadian period results from an organizational effect of androgen exposure during a post-pubertal sensitive period. Male degus were castrated before puberty and implanted with capsules filled with dihydrotestosterone (DHT), 17ß-estradiol (E2) or empty capsules at one of three ages: peri-puberty (2-7 months), post-puberty (7-12 months), or adulthood (14-19 months). Long-term exposure to DHT or E2 did not result in a shortened free-running circadian period when administered at 2-7 or 14-19 months of age. However, E2 treatment from 7 to 12 months of age decreased the free-running circadian period in castrated males. This result was replicated in a subsequent experiment in which E2 treatment was limited to 8-12 months of age. E2 treatment at 7-12 months of age had no effect on the free-running circadian period in ovariectomized females. Thus, there appears to be a post-pubertal sensitive period for sexual differentiation of the circadian system of degus, during which E2 exposure decreases the free-running circadian period in males. These data demonstrate that gonadal hormones can act during adolescent development to permanently alter the circadian system.

Role of gonadal hormones on mu-opioid-stimulated [³5S]GTPγS binding and morphine-mediated antinociception in male and female Sprague-Dawley rats.

RATIONALE: Male rats are more sensitive to morphine-mediated antinociception than female rats. A role for gonadal hormones in this sex difference has not been clearly defined. OBJECTIVES: To test the hypothesis that in vivo manipulation of gonadal hormones alters morphine-mediated G protein activation and leads to changes in morphine-mediated antinociception. METHODS: Adult male and female rats were gonadectomized and treated with either estradiol or testosterone in the females or testosterone in the male for up to 10 days. The ability of morphine and the peptidic mu-opioid agonist [D-Ala(2), N-MePhe(4), Gly-ol]-enkephalin (DAMGO) to stimulate [(35)S]GTPγS binding was measured in brain slices. In separate groups of identically treated rats, the antinociceptive response to morphine was determined using the warm-water tail-withdrawal assay. RESULTS: In the thalamus, morphine- and DAMGO-stimulated [(35)S]GTPγS binding was reduced by estradiol treatment of gonadectomized females compared to gonadectomized females treated with vehicle or testosterone. In the nucleus accumbens, the morphine-stimulated [(35)S]GTPγS binding was increased by estradiol treatment of gonadectomized females. In males, castration caused an increase in agonist-stimulated binding in the thalamus and a reduction in the amygdala compared with intact males. No significant changes were seen in mu-opioid agonist-stimulated [(35)S]GTPγS binding in other brain regions. There was no difference in antinociception following the systemic administration of morphine across the different hormonal manipulation conditions and the greater sensitivity of males was maintained irrespective of the treatment conditions. CONCLUSIONS: The modulation of mu-opioid receptor activation of G proteins by manipulation of sex hormones is region-specific and not reflected in antinociceptive responsiveness to morphine.

Changes in circadian rhythms during puberty in Rattus norvegicus: developmental time course and gonadal dependency.

During puberty, humans develop a later chronotype, exhibiting a phase-delayed daily rest/activity rhythm. The purpose of this study was to determine: 1) whether similar changes in chronotype occur during puberty in a laboratory rodent species, 2) whether these changes are due to pubertal hormones affecting the circadian timekeeping system. We tracked the phasing and distribution of wheel-running activity rhythms during post-weaning development in rats that were gonadectomized before puberty or left intact. We found that intact peripubertal rats had activity rhythms that were phase-delayed relative to adults. Young rats also exhibited a bimodal nocturnal activity distribution. As puberty progressed, bimodality diminished and late-night activity phase-advanced until it consolidated with early-night activity. By late puberty, intact rats showed a strong, unimodal rhythm that peaked at the beginning of the night. These pubertal changes in circadian phase were more pronounced in males than females. Increases in gonadal hormones during puberty partially accounted for these changes, as rats that were gonadectomized before puberty demonstrated smaller phase changes than intact rats and maintained ultradian rhythms into adulthood. We investigated the role of photic entrainment by comparing circadian development under constant and entrained conditions. We found that the period (τ) of free-running rhythms developed sex differences during puberty. These changes in τ did not account for pubertal changes in entrained circadian phase, as the consolidation of activity at the beginning of the subjective night persisted under constant conditions in both sexes. We conclude that the circadian system continues to develop in a hormone-sensitive manner during puberty.

Comparison of the antinociceptive response to morphine and morphine-like compounds in male and female Sprague-Dawley rats.

Male rats are more sensitive to the antinociceptive effects of morphine than female rats. This difference is seen across several rat strains using a variety of nociceptive stimuli. However, the literature in regard to sex differences in antinociceptive responses to mu-opioids other than morphine is less consistent. The present study was designed to examine whether there is a structure-activity rationale that determines which mu-opioids will show a differential antinociceptive response between male and female rats. A series of morphinans closely related in structure to morphine, namely, codeine, heroin, hydrocodone, hydromorphone, oxymorphone, and oxycodone, were examined for their antinociceptive activity in male and female Sprague-Dawley rats and compared with the structurally unrelated mu-opioid agonists methadone and fentanyl. Antinociception was measured by the warm-water tail-withdrawal assay. The results show that morphine is more potent in males compared with females > hydromorphone = hydrocodone = oxymorphone, but there was no observable sex difference in the antinociceptive potency of codeine, heroin, oxycodone, methadone, or fentanyl. The potency to stimulate guanosine 5'-O-(3-[35 S]thio)triphosphate ([35S]GTPgammaS) binding and binding affinity of the various morphinans was compared in rat glioma C6 cells expressing the rat mu-opioid receptor; relative efficacy was also compared by stimulation of [35S]GTPgammaS binding in slices of rat brain thalamus. The presence of a sex difference in antinociceptive responsiveness was not related to drug potency, efficacy, or affinity. Consequently, it is likely that differential metabolism of the opioid, possibly by glucuronidation, determines the presence or absence of a sex difference.

Comparison of the antinociceptive effect of acute morphine in female and male Sprague-Dawley rats using the long-lasting mu-antagonist methocinnamox.

Male rats are more sensitive than female rats to the antinociceptive action of morphine. The present study used age-matched (9-10 weeks old) male and female Sprague-Dawley rats to investigate whether this difference is due to variation in micro-opioid receptor binding and G protein activation. In the warm-water tail-withdrawal assay at both 50 degrees C and 55 degrees C, morphine was 2-3 times more potent in males than females. In contrast, micro-opioid receptor number and the binding affinity of the micro-opioid agonists morphine and DAMGO in membranes from whole brain, cortex, thalamus, and spinal cord were not different between males and females. Similarly, morphine and DAMGO stimulation of G protein, determined using GTPase and [(35)S]GTPgammaS binding assays, did not show a difference between the sexes. The long-lasting micro-opioid receptor antagonist methocinnamox (0.32 mg/kg), given 24 h prior to morphine, reduced micro-opioid receptor number by approximately 50% in thalamic and spinal cord tissue from female and male rats and reduced the antinociceptive potency of morphine. Pretreatment of male rats with 0.32 mg/kg methocinnamox reduced the antinociceptive potency of morphine to that observed in female rats expressing a full complement of micro-opioid receptors. However, with increasing pretreatment doses of methocinnamox, the maximal antinociceptive effect of morphine was decreased in females but not males. The results suggest that pathways downstream of the micro-opioid receptor and G protein are more efficient in male rats than in female rats such that there is a larger receptor reserve for morphine-mediated antinociception.