Distinct patterns of DeltaFosB induction in brain by drugs of abuse.

The transcription factor DeltaFosB accumulates and persists in brain in response to chronic stimulation. This accumulation after chronic exposure to drugs of abuse has been demonstrated previously by Western blot most dramatically in striatal regions, including dorsal striatum (caudate/putamen) and nucleus accumbens. In the present study, we used immunohistochemistry to define with greater anatomical precision the induction of DeltaFosB throughout the rodent brain after chronic drug treatment. We also extended previous research involving cocaine, morphine, and nicotine to two additional drugs of abuse, ethanol and Delta(9)-tetrahydrocannabinol (Delta(9)-THC, the active ingredient in marijuana). We show here that chronic, but not acute, administration of each of four drugs of abuse, cocaine, morphine, ethanol, and Delta(9)-THC, robustly induces DeltaFosB in nucleus accumbens, although different patterns in the core vs. shell subregions of this nucleus were apparent for the different drugs. The drugs also differed in their degree of DeltaFosB induction in dorsal striatum. In addition, all four drugs induced DeltaFosB in prefrontal cortex, with the greatest effects observed with cocaine and ethanol, and all of the drugs induced DeltaFosB to a small extent in amygdala. Furthermore, all drugs induced DeltaFosB in the hippocampus, and, with the exception of ethanol, most of this induction was seen in the dentate. Lower levels of DeltaFosB induction were seen in other brain areas in response to a particular drug treatment. These findings provide further evidence that induction of DeltaFosB in nucleus accumbens is a common action of virtually all drugs of abuse and that, beyond nucleus accumbens, each drug induces DeltaFosB in a region-specific manner in brain.

Inducible, brain region-specific expression of a dominant negative mutant of c-Jun in transgenic mice decreases sensitivity to cocaine.

Administration of cocaine induces the Fos family of transcription factors in the striatum, including the nucleus accumbens (NAc), a brain region important for the rewarding effects of addictive drugs. Several Fos proteins are induced acutely by cocaine, with stable isoforms of DeltaFosB predominating after chronic drug administration. However, it has been difficult to study the functional consequences of these Fos responses in vivo. Fos proteins heterodimerize with members of the Jun family to form active AP-1 transcription factor complexes. In the present study, we took advantage of this property and generated transgenic mice, using the tetracycline gene regulation system, that support the inducible, brain region-specific expression of a dominant negative mutant form of c-Jun (Deltac-Jun), which can antagonize the actions of Fos proteins. Expression of Deltac-Jun in the striatum and certain other brain regions of adult mice decreases their development of cocaine-induced conditioned place preference, suggesting reduced sensitivity to the rewarding effects of cocaine. In contrast, Deltac-Jun expression had no effect on cocaine-induced locomotor activity or sensitization. However, expression of Deltac-Jun in adult mice blocked the ability of chronic cocaine administration to induce three known targets for AP-1 in the NAc: the AMPA glutamate receptor subunit GluR2, the cyclin-dependent protein kinase Cdk5, and the transcription factor nuclear factor-kappaB (NFkappaB), without affecting several other proteins examined for comparison. Taken together, these results provide further support for an important role of AP-1-mediated transcription in some of the behavioral and molecular mechanisms underlying cocaine addiction.

Temporal effects of estrogen and progesterone on behavioral and endocrinological responses to acute cocaine administration.

Estrogen and progesterone have been postulated to play a key role modulating cocaine-induced behavioral and neurochemical activation in female rats. This study investigated the temporal relationship between estrogen and progesterone in the modulation of cocaine-induced behavioral alterations. Ovariectomized Fischer rats received s.c. injections of estradiol benzoate 48 hr prior to cocaine or saline treatment and one s.c. injection of progesterone concurrently or 1, 4, 20, 24, 30, 44 or 48 hr after estrogen treatment. Forty-eight hours after estrogen treatment rats received either a single i.p. injection of 15 mg/kg of cocaine or 0.9% saline. Overall, cocaine induced increases in locomotor behaviors (ambulatory and rearing activity). A bimodal interaction between estrogen and progesterone was observed in the modulation of all locomotor activities. A gradual increase in behaviors, which peaked when progesterone was administered 24 hr after estrogen was followed by an inhibition of both ambulatory and rearing activity when progesterone was administered for a shorter period of time. This estrogen and progesterone interaction was not observed in the modulation of cocaine-induced stereotypic activity. However, shorter administration of progesterone in relation to estrogen administration resulted in lowered benzoylecgonine plasma levels when compared to longer progesterone administration times. On the other hand, longer administration of progesterone (48 hr of estrogen and progesterone) caused increases in corticosterone levels in cocaine-treated rats. Thus, the temporal interaction between estrogen and progesterone in the regulation of cocaine metabolism and hypothalamic-pituitary-axis (HPA) activation do not completely correlate with that observed for locomotor behavioral activation. Taken together, these results suggest that temporal interactions between estrogen and progesterone may underlie some of the previously reported estrous cycle and sex effects on cocaine-induced behavioral and endocrinological alteration.

Endocrinological basis of sex differences in cocaine-induced behavioral responses.

Currently, 1.8 million Americans use cocaine, 30% of whom are females. Sex differences in the pattern of cocaine abuse may reside in neuroendocrinological modulations that affect the use of and/or dependence on cocaine. This review discusses sex differences in cocaine-induced behavioral and molecular alterations in the central nervous system, with emphasis on the role of endocrine responses in the neuronal modulations of this drug. Mechanisms and data supporting the role of the hypothalamic-gonadal axis in the modulation of cocaine-induced behavioral and molecular alterations are also provided.

Ovarian hormones modulate cocaine-induced locomotor and stereotypic activity.

Interactions between ovarian hormones and cocaine may underlie gender and estrous cycle differences in cocaine-induced behavioral and neurochemical alterations. The aim of this study was to further understand how ovarian hormones modulate cocaine-induced behavioral alterations. Ovariectomized rats received acute or chronic saline or cocaine (15 mg/kg i.p.) administration and were further subdivided into one of four hormone-treatment conditions: cholesterol (vehicle-control), estrogen, progesterone, or estrogen + progesterone. Overall, acute and chronic cocaine administration increased all locomotor measurements (total locomotor, ambulatory, and rearing counts). Estrogen administration augmented cocaine-induced increases in ambulatory and rearing activity. After chronic cocaine administration, rats in the vehicle-control group developed behavioral tolerance (exhibited by a decrease in activity) in rearing and ambulatory activity. Estrogen replacement not only prevented the development of tolerance in ambulatory and rearing activities, but also enhanced total locomotor activity (sensitization) in response to chronic cocaine. Progesterone treatment did not alter the behavioral responses to acute or chronic cocaine administration. Estrogen + progesterone-treated animals had higher counts of locomotor activity in response to chronic cocaine than did vehicle-control or progesterone-treated rats. In stereotypic behaviors, the different hormonal treatments did not affect activity in cocaine- or saline-treated rats after acute or chronic drug administration. Plasma levels of cocaine did not change after different hormonal treatments. Interestingly, animals' coadministered chronic cocaine and estrogen had higher levels of corticosterone than did nonestrogen cocaine-treated rats. Thus, it is likely that alterations in HPA activation may underlie the observed behavioral differences. In summary, these data suggest that there are interactions between ovarian hormones and cocaine-induced behavioral alterations in female rats, and they extend previous results by showing that estrogen and progesterone affect the development of sensitization.

Vendor differences in cocaine-induced behavioral activity and hormonal interactions in ovariectomized Fischer rats.

Contradictory effects of ovarian hormone on cocaine-induced behaviors have been reported in ovariectomized Fischer rats. To determine if these discrepancies are based on where the rats were purchased, Charles River Laboratories and Taconic Fischer rats were randomly assigned to either cocaine (15 mg/kg, intraperitoneal) or saline treatment; and one of four hormone-pretreatment subgroups: vehicle, estrogen, progesterone or estrogen + progesterone. Vendor differences were observed in cocaine-induced locomotor activities; overall, Taconic rats demonstrated less locomotor activity than Charles River rats. Furthermore, vendor differences in ambulatory activity were also observed after steroid replacement treatment. In Charles River rats, estrogen + progesterone co-administration suppressed cocaine-induced increases in ambulatory activity when compared to other hormone-treated groups given cocaine. In contrast, Taconic rats showed an increase in ambulations after this drug/hormone treatment. Vendor differences were also observed in steroid effects on cocaine-induced rearing activity, where estrogen + progesterone and cocaine caused an increase in rearing in Charles River rats, but not in Taconic rats. No differences between the vendors were observed in saline- or cocaine-treated animals' stereotypic activity. Vendor differences in cocaine-induced locomotor activity were not due to differences in cocaine metabolism, as no differences in plasma levels of benzoylecgonine were observed. Interestingly, Taconic animals had overall higher plasma levels of corticosterone than Charles River rats. Thus, intrinsic differences between different lines of Fischer rats may affect the outcome of ovarian hormone interactions in cocaine-induced behavioral alterations.

Sex differences in cocaine-induced behavioral sensitization.

To further understand how sex differences affect the development and maintenance of sensitization, 48 adult Fischer rats (24 female and 24 male) received chronic administration (14 days) of cocaine (15 mg/kg, i.p.) or saline or a challenge dose (7 days after chronic cocaine administration). Sex differences were observed in the development and maintenance of cocaine-induced total locomotor, ambulatory and rearing activity. Although, overall cocaine administration increased stereotypic activity in both male and female rats, female rats had significantly higher stereotypic activity than male rats across the three behavioral test days (1, 7 and 14). Female rats had statistically significant higher benzoylecognine levels after acute cocaine administration than male rats. However, no differences between male and female rats in benzoylecognine plasma levels were observed after chronic and challenge doses of cocaine administration. Interestingly, after acute and challenge cocaine administration, corticosterone levels were significantly higher in female rats when compared to male rats. This study confirms previous reports that there are sex differences in the behavioral response to cocaine. Moreover, this study expands previous studies by demonstrating that sex differences occur in only certain aspects of cocaine-induced behavioral activation and the development and maintenance of cocaine-induced behavioral sensitization.

Progesterone and cocaine administration affect serotonin in the medial prefrontal cortex of ovariectomized rats.

Due to the hypothetical role of ovarian hormones, estrogen and progesterone, in cocaine-induced behavioral activity and self-administration, this study investigated the effects of cocaine, estrogen, and progesterone administration on monoamine levels in the medial prefrontal cortex of ovariectomized hormone-treated rats. Rats were given either 'binge' cocaine or saline, and one of four hormone treatments: vehicle, estrogen, progesterone, or estrogen+progesterone. The co-administration of progesterone and cocaine resulted in increased levels of serotonin when compared to saline-treated controls and cocaine-treated animals in the other hormone-treatment groups. Further, progesterone-treated rats had higher levels of 5-HIAA than vehicle or estrogen-treated rats. Although levels of dopamine, DOPAC, and homovanillic acid were decreased after cocaine, these alterations failed to reach significance. These results show an interaction between the endocrine environment and cocaine-induced alterations in serotonin system in the medial prefrontal cortex. Thus, these changes may contribute to previously reported gender and estrous cycle differences in behavioral responses to cocaine.

Cocaine affects progesterone plasma levels in female rats.

Female Fischer rats injected with cocaine in a "binge" pattern (15 mg/kg, IP, three times a day, at 1-h intervals) for 1 day had significantly higher levels of progesterone than saline-treated controls (p < 0.001). When analyzed by the stage of the estrous cycle, animals in proestrus showed significantly higher cocaine-induced progesterone plasma levels than those in other stages of the cycle (p < 0.01). Progesterone plasma levels were also increased after a single dose of cocaine (15 mg/kg). However, 3 h postinjection progesterone plasma levels had returned to normal. Thus, cocaine modulation of progesterone plasma levels appears to be an acute effect. In ovariectomized rats pretreated with estrogen, progesterone, or estrogen + progesterone, no significant differences were observed in progesterone plasma levels after acute "binge" pattern cocaine administration. Thus, acute cocaine induced increases in progesterone plasma levels in intact female rats are probably due to an increase in secretion rates of progesterone rather than an acceleration of its biotransformation. Due to the profound effects of progesterone in the modulation of CNS plasticity, the modulation of progesterone plasma level by cocaine may have implications for reproductive processes and neuronal functions of women. Moreover, cocaine may affect the progesterone levels in women utilizing progesterone-based contraception or steroid replacement treatment after menopause.

Ovarian hormone replacement affects cocaine-induced behaviors in ovariectomized female rats.

To determine whether cocaine-induced behavioral alterations are modulated by ovarian hormones, ovariectomized rats were randomly assigned to one of two drug treatment conditions: "binge" cocaine (three 15-mg/kg intraperitoneal (ip) injections, 1 h apart) or saline administration; and four hormone pretreatment sub-groups: vehicle control, estrogen, progesterone, or estrogen+progesterone. Cocaine-treated animals displayed more locomotor activity than saline-treated animals and locomotor activity was higher after the third injection than after the first two injections. When analyzed according to hormone group, the administration of estrogen+progestrone suppressed cocaine-induced locomotion after the first injection; this effect was significant when compared to estrogen-pretreated animals. While in each condition cocaine-treated animals displayed significantly higher stereotypic activity than saline-treated animals, in the estrogen+progesterone replacement group, there was more activity after the second injection of cocaine than after the first. Interestingly, animals in the estrogen+progesterone group had significantly lower plasma levels of the cocaine metabolite, benzoylecgonine, than animals in the progesterone or estrogen groups. These results extend our earlier findings in the intact female rat, which suggest an interaction between the endocrine environment, cocaine metabolism, and cocaine-induced behaviors. These effects may underlie reported sex and estrous cycle differences in cocaine-induced behavioral activity.