The nucleus accumbens as a site of action for rewarding properties of testosterone and its 5alpha-reduced metabolites.

Testosterone (T)'s positive hedonic effects may be mediated by actions of its metabolites, dihydrotestosterone (DHT) or 3alpha-androstanediol (3alpha-diol), in the nucleus accumbens (NA). In Experiment 1, adult, intact, male rats were systemically administered 1 mg of T, DHT, 3alpha-diol or vehicle, at different time points to examine concentrations of androgens in the NA. Rats administered 3alpha-diol had significantly increased concentrations of 3alpha-diol in the region of the brain encompassing the NA. These data are consistent with previous data from our laboratory demonstrating that 3alpha-diol elicits a conditioned place preference (CPP) more effectively than either T or DHT, when administered systemically. In Experiment 2, rats received implants of T, DHT or 3alpha-diol to the NA immediately prior to placement in the CPP apparatus on conditioning days. Implants of T, DHT or 3alpha-diol, but not vehicle, significantly increased time spent on the non-preferred side of the chamber on the test day. This effect was only produced by androgenic stimulation of the shell of the NA and not the core of the NA. Thus, androgen regimens we have previously found to enhance CPP produced the greatest increases in 3alpha-diol concentrations in the NA region and direct implants of T, DHT or 3alpha-diol to the shell, but not the core, of the NA enhanced CPP. These data are consistent with the hypothesis that the hedonic effects of T may be due to actions of its metabolites in the NA.

The testosterone metabolite and neurosteroid 3alpha-androstanediol may mediate the effects of testosterone on conditioned place preference.

The abuse of androgens may be related to their ability to produce positive, hedonic interoceptive effects. Conditioned Place Preference (CPP) has been used in many experiments to examine hedonic effects of drugs. This review is focused on studies from our laboratory that utilized CPP to examine potential positive hedonic effects of testosterone (T), and its androgenic metabolite dihydrotestosterone (DHT), and its metabolite 3alpha-androstanediol (3alpha-diol). We hypothesized that administration of a high concentration of 3alpha-diol would produce a CPP, pharmacological concentrations of plasma androgens, and alter androgen receptors (AR) and the function of GABA(A)/benzodiazepine receptor complexes (GBR). In our studies, we observed that systemic 3alpha-diol (1.0 mg/kg) prior to exposure to the non-preferred side of a CPP chamber significantly increased preference for the non-preferred side of the chamber compared to baseline preference and homecage controls. Furthermore, administration of T, DHT, or 3alpha-diol increased levels of these androgens, decreased ARs (decreased seminal vesicle weight and intrahypothalamic AR) and GBR function (decreased GABA-stimulated chloride influx in cortical synaptoneurosomes, and muscimol binding in the hippocampus compared to control groups). With systemic administration of 3alpha-diol that enhanced CPP, concentrations of 3alpha-diol were increased in the nucleus accumbens (NA). Central implants of T, DHT, or 3alpha-diol to the NA also produced a CPP compared to baseline preference and vehicle controls. These data indicate that systemic 3alpha-diol is more effective at enhancing CPP and increasing circulating 3alpha-diol levels than is T or DHT and that central administration of 3alpha-diol to the NA can condition a place preference. These findings indicate that 3alpha-diol produces positive hedonic effects and suggest that T's variable effects on CPP may be due in part to T's metabolism to 3alpha-diol.

The testosterone metabolite and neurosteroid 3alpha-androstanediol may mediate the effects of testosterone on conditioned place preference.

Testosterone (T) and pregnane neurosteroids can enhance conditioned place preference (CPP). The present experiment examined CPP produced by T and its androgenic metabolite dihydrotestosterone (DHT) and 3alpha-Androstanediol (3alpha-diol; an androstane neurosteroid). Administration of 3alpha-diol (>DHT>T) to intact male Long-Evans rats, 1.0 mg daily for six days, 30 min prior to exposure to the non-preferred side of the CPP chamber significantly increased preference for the non-preferred side of the chamber compared to that seen in home cage controls. Levels of circulating 3alpha-diol were increased significantly in 3alpha-diol>DHT>T-administered rats, compared to rats that had vehicle administered or androgen-administration discontinued. Androgen administration decreased seminal vesicle weight and intrahypothalamic androgen receptor (AR) binding compared to that seen in rats that had vehicle administered or androgen-administration discontinued. Testosterone, DHT, and 3alpha-diol decreased GABA-stimulated chloride influx in cortical synaptoneurosomes, and muscimol binding in the hippocampus compared to that seen in rats with vehicle administered or that had androgen-administration discontinued. These data indicate that administration of 3alpha-diol is more effective at enhancing CPP and increasing circulating 3alpha-diol levels than is DHT or T administration, and that all of the androgen regimens employed decreased peripheral and hypothalamic androgen receptor binding and cortical and hippocampal GABA(A) receptor function. Hence, whether the effects of 3 alpha-diol on CPP are mediated by differential actions at ARs or GABA(A) receptors in particular brain regions needs to be determined.

Postpartum aggression in mice: the role of ovarian hormones.

The development of aggression during the immediate postpartum period was investigated in Rockland-Swiss albino female mice. Aggression was low immediately following delivery and increased with advancing lactation. The majority of females delivered during the dark cycle, however, whether females delivered or were tested during the light or dark cycle did not influence aggression. Females ovariectomized on Gestation Day 18 displayed aggression sooner than sham-operated controls. Estrogen treatment restored typical postpartum docility in prepartum ovariectomized females. Ovariectomy also increased the number of nursing bouts, but not total nursing time, when compared to sham-operated females. These findings suggest that peripartum estrogen stimulation, directly or through influencing nursing activity, prevents aggression in females immediately postpartum.

Investigation of anabolic steroids in two taste aversion paradigms.

The aversive effects of estradiol have been studied in two different taste aversion paradigms. A similar investigation was undertaken for the anabolic-androgenic steroids, nandralone and testosterone cypionate, using Rockland-Swiss mice. Experiments 1 and 2 used the brief exposure of a novel saccharin solution as the conditioned stimulus for taste aversion learning, and showed that anabolic steroids (1 mg) do not induce taste aversions. Instead, these hormones induced a small non-contingent increase in saccharin preference. Experiment 3 showed that daily nandralone administration (1 mg/day) had a greater anabolic effect than the same dose of testosterone cypionate. Experiment 4 paired the continuous exposure to a novel diet with daily nandralone injections, and showed that steroid treatment increased intake of the novel diet. When the novel diet was subsequently presented with the familiar diet in a two-choice preference test, there was no indication that an aversion was conditioned to the novel target diet. On the contrary, nandralone treatment significantly increased the preference for the novel diet. These experiments show that anabolic-androgenic steroids do not have aversive effects in mice, and that they may have positive consequences.

Estradiol benzoate can function as an unconditioned stimulus in a conditioned taste aversion paradigm.

The extent to which gonadal steroid hormones can serve as unconditioned stimuli in a conditioned taste aversion paradigm was examined in Rockland-Swiss albino mice. With saccharin serving as the conditioned stimulus, subcutaneously injected estradiol benzoate, but not progesterone or testosterone propionate, was found to be a potent unconditioned stimulus in both male and female mice. Dose-response effects were also observed; increasing dosages of estradiol benzoate led to increasingly stronger conditioned aversions in both males and females. The aversion detected in males was more resistant to extinction than that seen in females. Prepubertal gonadectomy reversed the sex-dependent effects of estradiol benzoate in learned aversions in adulthood; castration of males promoted the extinction process, whereas ovariectomy of females retarded extinction. The results may be useful for our understanding of the mechanisms involved in conditioned taste aversion learning as well as a wide array of hormone-dependent behavioral responses.

Genotype modulates the aggression-promoting quality of progesterone in pregnant mice.

During late pregnancy, female mice of the DBA/2J inbred strain are more likely to exhibit aggressive behavior toward a standard stimulus intruder male than C57BL/6J females. This strain difference can not be accounted for by differences in circulating levels of progesterone (P) since pregnant DBA/2J and C57BL/6J females exhibit similar patterns of the steroid throughout pregnancy. Upon receiving subcutaneously implanted Silastic capsules containing P, virgin DBA/2J mice are more likely than virgin C57BL/6J to respond to the steroid by exhibiting aggression. Strain differences in the aggressive behavior exhibited by pregnant mice may be related to genotype-based variation in central neural tissue sensitivity to P.

Suckling stimulation modulates the maintenance of postpartum aggression in mice.

Previous work in female mice has shown that suckling stimulation is essential for the initiation of male-directed aggression during the immediate postpartum period. The present work examined the importance of this stimulus for the maintenance of aggression during lactation. Surgical excision of nipples (thelectomy) on Postpartum Day 5 accelerates the decline in aggression that ordinarily occurs during the final third of lactation (Postpartum Days 15-21), but does not disrupt the behavior during the middle third of lactation (Postpartum Days 6-12). Thus, both the initiation of aggression during the early postpartum period and the maintenance of the behavior during late lactation are controlled by suckling from young.

Prenatal stress alters maternal aggression in mice.

Prenatal stress (heat and restraint) reduced pregnancy-induced and elevated postpartum aggression in Rockland-Swiss (R-S) albino female mice. Though prenatally-stressed females were indistinguishable from control animals with respect to parental behavior during the virgin state, the former displayed slightly lower levels of nestbuilding during early pregnancy, and delivered slightly more male offspring at parturition. The young born to prenatally-stressed females exhibited deficits in birth weights and body weight gain in contrast to pups delivered by control females. The anogenital distances of prenatally-stressed females were shorter than those of control females, suggesting that alterations in fetal testosterone exposure may be responsible for disruptions in behavior and reproduction.

Genotype modulates prenatal stress effects on aggression in male and female mice.

Prenatal stress (heat and restraint) significantly increased postpartum aggression (proportion of animals fighting and/or the intensity of the behavior) in C57BL/6J female mice and reduced the behavior in DBA/2J females. For intermale aggression, prenatal stress increased the behavior (intensity of aggression) in C57BL/6J males but did not affect aggressive behavior in DBA/2J animals. Infanticidal behavior (the killing of young) exhibited by male mice was not influenced by prenatal stress in either strain. Relative anogenital distance measurements in neonates at birth did not serve as a reliable predictor of strain variation in prenatal stress effects. Prenatal stress did not influence this measure of prenatal androgen exposure in DBA/2J or C57BL/6J females. For males, prenatal stress elevated relative anogenital distance in C57BL/6J mice and decreased this measure in DBA/2J animals. Prenatal stress effects on aggressive behavior in male and female mice therefore depend upon genotype. Strain-dependent differences may be modulated by differences in endocrine reactivity to prenatal stress/and or differential central neural tissue sensitivity to hormones.

Adrenal modulation of parental behavior in neonatally gonadectomized C57BL/6J male mice.

C57BL/6J male mice ordinarily kill neonatal mouse pups even if they are rendered androgen deficient by neonatal castration. Experiment 1 showed that adrenalectomy during adult life significantly decreased the tendency of neonatally gonadectomized males to kill newborns. Experiment 2 demonstrated that testosterone exposure (via silastic implants) during adult life prevented the effects of adrenalectomy in that killing was elevated and retrieving was reduced in neonatally gonadectomized males. Adrenal androgens may be responsible for maintaining killing behavior in neonatally castrated C57BL/6J male mice.

Prior intrauterine position influences body weight in male and female mice.

In two longitudinal studies, intrauterine location of male and female Rockland-Swiss mice relative to fetuses of the same and opposite sex dramatically influenced body weight. In one study, body weight of males and females that were located in utero between two female fetuses (OM animals) or between two male fetuses (2M animals) was assessed from birth to the time of weaning (25 days of age). The body weights of 2M females were indistinguishable from those of OM and 2M males on all but a few of the 25 postnatal days of life. Also, 2M females were significantly heavier than OM females from Postnatal Day 6 onward, and 2M males weighed significantly more than OM males from Postnatal Day 19 onward. In a second study, food intake and body weight of animals from different intrauterine locations were examined from 25 to 120 days of age. Regardless of prior intrauterine position, males were always heavier than females. However, prior in utero location modulated body weight in that 2M females were significantly heavier than OM females and 2M males were reliably heavier than OM males. Intrauterine position effects were observed in the absence of any appreciable influence of this variable on levels of food intake. Taken together, the results suggest that prior in utero location may influence metabolic set points involved in the regulation of body weight and fat storage.

Mice: progesterone stimulates aggression in pregnancy-terminated females.

Three experiments were conducted in order to assess the role of progesterone (P) in the aggressive behavior displayed by late pregnant Rockland-Swiss mice toward adult male intruders. In Experiment 1, hysterectomy on the 15th day of gestation reduced the aggressive behavior normally displayed by pregnant mice toward male intruders. In Experiment 2, Silastic implants of P stimulated aggression in hysterectomized mice but did not fully restore the behavior to the level of fighting normally observed in pregnant animals. In Experiment 3, aggressive behavior in P-treated hysterectomized animals was inhibited by simultaneous exposure to estradiol (E). Also, treatment with E alone did not stimulate aggression in hysterectomized mice. While pregnancy-induced aggression is promoted by P, other neuroendocrine factors may act in concert with the steroid to fully stimulate aggression in gravid mice.

Mice: postpartum aggression is elevated following prenatal progesterone exposure.

Pregnant Rockland-Swiss (R-S) mice were injected with sesame oil or 250 or 500 micrograms of progesterone (P) on Days 12 through 16 of gestation and the postpartum aggressive behavior of their female offspring was examined in adulthood. Both doses of P significantly increased the intensity of aggression (number of attacks) exhibited by the female offspring toward an adult male intruder. The low dose of P also produced significant increases in relative anogenital distance. These effects were seen in the absence of any effects on body weight at birth or in adulthood, or on reproductive performance. The findings support previous research, in both animals and humans, showing that prenatal brain differentiation and subsequent behavior are masculinized by prenatal exposure to progesterone.

Intrauterine contiguity influences regulatory activity in adult female and male mice.

Female mice located in utero between two female fetuses exhibited higher levels of locomotor activity in adulthood than did females located between two male fetuses. Male mice, which were less active than females, also were influenced by intrauterine contiguity. Males located in utero between two female fetuses were more active than males which resided between two male fetuses. These results indicate that intrauterine position influences behaviors involved in the maintenance of metabolic homeostasis.