The role of the androgen receptor in anabolic androgenic steroid-induced aggressive behavior in C57BL/6J and Tfm mice.

Humans self-administer anabolic androgenic steroids (AAS) at superphysiological doses for the purpose of building muscle mass and enhancing physique whereas considerably lower doses of AAS are prescribed in the clinic to treat a variety of disorders. A number of studies have demonstrated that individual AAS influence aggressive behavior in rats and mice, but few studies have examined the aggression-enhancing effects of combinations of AAS. Using the resident-intruder paradigm, Experiment 1 determined whether a cocktail of commonly abused AAS increased aggressive behavior in gonadally-intact male C57BL/6J mice and examined whether the androgen receptor (AR) was involved. Mice given either AAS cocktail or the cocktail and the AR antagonist, flutamide, for 6 weeks were subject to three weekly tests in which the percentage of mice that fought, the latency to initiate an aggressive event and the number of aggressive events per 5-min-fight session were recorded. In C57BL/6J mice, 6 weeks of AAS administration increased the likelihood of fighting, however, within the subset of mice that engaged in aggression, AAS did not specifically modulate the latency to fight or the number of aggressive events per fight. In addition, co-administration of flutamide only slightly altered the likelihood that mice given AAS will initiate a fight. Experiment 2 examined the aggression-promoting effects of AAS in gonadally-intact adult testicular feminization mutant (Tfm) mice, which are deficient in functional ARs. Overall, fewer Tfm mice compared to C57BL/6J mice fought in both drug conditions (vehicle or AAS). Taken together, these data suggest that given the presence of AR during development, AAS enhance adult male aggression in C57BL/6J mice through AR-independent and AR-dependent pathways. In contrast, in adult Tfm mice, the likelihood of AAS-enhanced aggression in adulthood is significantly reduced.

An intact medial preoptic area is necessary for zaprinast to modulate paced mating behavior in female rats.

The present study examined the interaction between the regulation of paced mating behavior by the medial preoptic area (mPOA) and by the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway, as modulated by zaprinast, a phosphodiesterase inhibitor. Rats receiving mPOA or sham lesions were tested for paced mating behavior. Subsequently, rats were treated with zaprinast (3 mg/kg) before a second paced mating test. The expected lengthening of contact-return latencies following intromissions and ejaculations was observed in rats with mPOA lesions relative to rats with sham lesions. In addition, rats with sham lesions responded to zaprinast with a lengthening of contact-return latency following ejaculation. Contact-return latencies did not change in response to zaprinast in rats with mPOA lesions. These results demonstrate that the alterations in paced mating behavior observed in rats with mPOA lesions persist despite manipulation of the nitric oxide-cyclic guanosine monophospate pathway.

The display of paced mating behavior in a rat model of endometriosis.

Endometriosis is a disorder associated with chronic pelvic pain and ill effects on women's sexual health. The present study examined the effects of pelvic endometriotic implants on the display of paced mating behavior in female rats. Approximately 2 months after the surgical induction of endometriosis, rats were tested for paced mating behavior during proestrus (Experiment 1) or after bilateral ovariectomy and hormone replacement (Experiment 2). Although endometriotic implants were confirmed at autopsy, rats with surgical endometriosis in both experiments exhibited normal patterns of paced mating behavior. The positive relationship between implant material and contact-return latency following ejaculation in Experiment 2 suggests that the sensitivity to vigorous mating stimulation may be influenced by endometriosis.

Genitosensory nerve modulation of paced mating behavior: evidence for pelvic, but not hypogastric, nerve influence.

The pelvic nerve is known to play a role in the behavioral and neurochemical responses exhibited during paced mating behavior. The present study extended the analysis of the contribution of the genitosensory nerves to the display of paced mating behavior to include bilateral hypogastric nerve transection, bilateral pelvic nerve transection, or transection of both the hypogastric and pelvic nerves. Rats with pelvic nerve transection were less likely to exit the male compartment, took longer to exit the male compartment following intromissions, and returned to the male more quickly following intromissions compared to rats with an intact pelvic nerve. In contrast, hypogastric nerve transection alone did not affect paced mating and had no modulating effect on the paced mating behavior of rats with pelvic nerve transection. Our results support the view that key aspects of paced mating behavior are modulated by signals transmitted via the pelvic nerve, without any discernable contribution from the hypogastric nerve.

Androgen inhibition of sexual receptivity is modulated by estrogen.

Sexual receptivity induced in ovariectomized rats by the long-term administration of estradiol benzoate (EB) can be inhibited by concurrent administration of androgens. Experiment 1 examined the role of time course and dose of androgens in the inhibition of estrogen-induced sexual receptivity. Ovariectomized rats were treated with EB (2.0 microg per rat per day) for 6 days and tested for sexual receptivity (Test Day I). EB treatment continued for 15 days concomitant with daily administration of one of three doses of dihydrotestosterone propionate (DHTP; 7.5, 0.75, 0.075 mg/kg) or 3α-androstanediol (3α-Adiol; 3.75, 1.0, 0.375 mg/kg). Four tests for sexual receptivity were conducted on days 3, 6, 14, and 15 of the androgen/vehicle treatment period (Test Days II-V). On Day 15 (Test Day V), the rats received progesterone (1.0 mg per rat) 4 h before testing. Using the same experimental design, Experiment 2 examined the effect of increasing the dose of estrogen on the androgenic inhibition of sexual receptivity. Ovariectomized rats were treated with one of two doses of EB (2.0 or 10.0 microg per rat per day) concomitant with daily administration of DHTP (7.5 mg/kg) or 3α-Adiol (3.75 mg/kg). In Experiment 1, the highest doses of both DHTP and 3α-Adiol significantly inhibited estrogen-induced sexual receptivity. Data from Experiment 2 indicate that the inhibitory effects of DHTP but not 3α-Adiol can be moderated by an increased dose of EB.

Paced mating behavior persists in rats with vaginocervical Lidocaine.

The present study tested whether the topical application of a local anesthetic (Lidocaine) to the vaginocervical region altered the pattern of paced mating behavior displayed by gonadectomized, hormone-primed female rats. Both rats receiving Lidocaine and rats receiving vehicle exhibited the expected lengthening of contact-return latency as the intensity of the mating stimuli increased (mount

Lesions of the medial preoptic area interfere with the display of a conditioned place preference for vaginocervical stimulation in rats.

The present study examined the effect of ibotenic acid lesions of the medial preoptic area (mPOA) on the expression of a conditioned place preference (CPP) for vaginocervical stimulation. Rats with bilateral lesions of the mPOA failed to display the CPP for vaginocervical stimulation shown by rats with sham or incomplete lesions. These findings provide additional support for the role of the mPOA in the neural circuitry underlying the reinforcing effects of female sexual behavior and raise the possibility that the altered pattern of approach and withdrawal behavior observed following lesions of the mPOA may be attributable in part to a diminution of the reinforcing effects of vaginocervical stimulation.

Conditioned place preference for mating is preserved in rats with pelvic nerve transection.

Female rats exhibit a conditioned place preference (CPP) for a context paired with mating. The present experiment tested the hypothesis that the activation of the pelvic nerve mediates the reinforcing effects of mating for female rats. Rats underwent bilateral pelvic nerve or sham transection and then received paced mating, nonpaced mating, or the control treatment during a CPP procedure. Pelvic nerve transection did not affect the CPP for paced or nonpaced mating. In tests of paced mating behavior, contact-return latencies following intromissions were significantly shorter in rats with pelvic nerve transection than they were in rats with sham transections. These results show that the pathway conveying the reinforcing effects of mating stimulation does not depend on the integrity of the pelvic nerve, but that activation of the pelvic nerve contributes to the display of paced mating behavior.

Life without thyroxine to 3,5,3'-triiodothyronine conversion: studies in mice devoid of the 5'-deiodinases.

Considerable indirect evidence suggests that the type 2 deiodinase (D2) generates T(3) from T(4) for local use in specific tissues including pituitary, brown fat, and brain, whereas the type I deiodinase (D1) generates T(3) from T(4) in the thyroid and peripheral tissues primarily for export to plasma. From studies in deiodinase-deficient mice, the importance of the D2 for local T(3) generation has been confirmed. However, the phenotypes of these D1 knockout (KO) and D2KO mice are surprisingly mild and their serum T(3) level, general health, and reproductive capacity are unimpaired. To explore further the importance of 5'deiodination in thyroid hormone economy, we used a mouse devoid of both D1 and D2 activity. In general, the phenotype of the D1/D2KO mouse is the sum of the phenotypes of the D1KO and D2KO mice. It appears healthy and breeds well, and most surprisingly its serum T(3) level is normal. However, impairments in brain gene expression and possibly neurological function are somewhat greater than those seen in the D2KO mouse, and the serum rT(3) level is elevated 6-fold in the D1/D2KO mouse but only 2-fold in the D1KO mouse and not at all in the D2KO mouse. The data suggest that whereas D1 and D2 are not essential for the maintenance of the serum T(3) level, they do serve important roles in thyroid hormone homeostasis, the D2 being critical for local T(3) production and the D1 playing an important role in iodide conservation by serving as a scavenger enzyme in peripheral tissues and the thyroid.

Zaprinast, a phosphodiesterase type-5 inhibitor, alters paced mating behavior in female rats.

Nitric oxide (NO) is the primary mediator of blood flow in female genital tissues and drugs that enhance the activity of nitric oxide, such as phosphodiesterase type-5 (PDE-5) inhibitors, increase vaginal blood flow in anesthetized rats. The goal of the present study was to test the effects of one PDE-5 inhibitor, zaprinast, on the display of sexual behaviors in gonadectomized, estrogen- and progesterone-treated female rats. Experiment 1 demonstrates that zaprinast alters paced mating behavior by lengthening the contact-return latency to ejaculation; there is a significant relationship between dose of zaprinast (range 1.5-6 mg/kg) and contact-return latency to ejaculation. Experiment 2 illustrates that zaprinast has no effect on preference for an intact male as measured in a No Contact partner preference test. Rats receiving zaprinast tend to exhibit reduced locomotor activity in both experiments. Collectively, these findings demonstrate that modulation of the NO-cGMP pathway using a PDE-5 inhibitor alters the display of paced mating behaviors in rats.

Artificial vaginocervical stimulation induces a conditioned place preference in female rats.

Female rats express a conditioned place preference (CPP) for a context paired with mating. During a mating encounter, the female rat is exposed to several different types of stimuli, including, but not limited to, vaginocervical stimulation and social contact. The present experiment tested the hypothesis that two components of the mating interaction, vaginocervical stimulation or social contact, each induce a CPP in female rats. During conditioning rats received nonpaced mating, artificial vaginocervical stimulation, social interaction or a control treatment. Rats expressed a CPP for the context paired with nonpaced mating or artificial vaginocervical stimulation whereas social interaction and the control treatment did not induce a CPP. The present findings highlight the important role that vaginocervical stimulation plays in the reinforcing effects of mating in female rats.

Thyroid hormone homeostasis and action in the type 2 deiodinase-deficient rodent brain during development.

Considerable indirect evidence suggests that the type 2 deiodinase (D2) generates T3 from T4 for local use in specific tissues such as pituitary, brown fat, and brain, and studies with a D2-deficent mouse, the D2 knockout (D2KO) mouse, have shown this to be the case in pituitary and brown fat. The present study employs the D2KO mouse to determine the role of D2 in the developing brain. As expected, the T3 content in the neonatal D2KO brain was markedly reduced to a level comparable with that seen in the hypothyroid neonatal wild-type mouse. However, the mRNA levels of several T3-responsive genes were either unaffected or much less affected in the brain of the D2KO mouse than in that of the hypothyroid mouse, and compared with the hypothyroid mouse, the D2KO mouse exhibited a very mild neurological phenotype. The current view of thyroid hormone homeostasis in the brain dictates that the T3 present in neurons is generated mostly, if not exclusively, from T4 by the D2 in glial cells. This view is inadequate to explain the findings presented herein, and it is suggested that important compensatory mechanisms must be in play in the brain to minimize functional abnormalities in the absence of the D2.

Female rats exhibit a conditioned place preference for nonpaced mating.

Paced, but not nonpaced, mating behavior is reported to induce a conditioned place preference (CPP) in female rats. Contrary to these previous findings, Experiment 1 showed that female rats that received 15 intromissions from a single male rat during each of five conditioning sessions exhibited a CPP for the compartment associated with mating when the intromissions were delivered via a paced or nonpaced paradigm. Experiment 2 demonstrated that nonpaced mating induced a CPP when a single male delivered the 15 intromissions but not when the male was replaced following ejaculation and a new male allowed to complete the requisite number of intromissions. These findings invite reevaluation of the reinforcing aspects of mating behavior in female rats.

Stimulus animal characteristics do not modulate the expression of partner preference by female rats.

The stimulus animals used in tests of partner preference in female rats vary. To test the hypothesis that the alternative stimulus animal modulates the preference for an intact male, gonadectomized (GDX) female rats received estradiol benzoate plus progesterone or the oil vehicle and were tested for partner preference with either an intact male paired with a GDX hormone-primed female or an intact male paired with a GDX male. Rats were tested under conditions that limited physical contact (No Contact) or allowed sexual interaction (Contact). Stimulus animal condition was not a primary determinant of partner preference. In contrast, contact condition and hormone treatment modulated preference, as well as activity levels and the display of proceptive behaviors. Our findings demonstrate that the characteristics of the alternative stimulus animals tested here do not play a significant role in modulating partner preference in female rats.

Sex- and age-specific effects of anabolic androgenic steroids on reproductive behaviors and on GABAergic transmission in neuroendocrine control regions.

Illicit use of anabolic androgenic steroids (AAS) has become a prevalent health concern not only among male professional athletes, but, disturbingly, among a growing number of women and adolescent girls. Despite the increasing use of AAS among women and adolescents, few studies have focused on the effects of these steroids in females, and female adolescent subjects are particularly underrepresented. Among the hallmarks of AAS abuse are changes in reproductive behaviors. Here, we discuss work from our laboratories on the actions of AAS on the onset of puberty and sexual behaviors in female rodents, AAS interactions and sex- and age-specific effects of these steroids on neural transmission mediated by gamma-aminobutyric acid receptors within forebrain neuroendocrine control regions that may underlie AAS-induced changes in these behaviors.

Ibotenic acid lesions of the medial preoptic area disrupt the expression of partner preference in sexually receptive female rats.

The present study evaluated the effects of ibotenic acid lesions of the medial preoptic area (mPOA) on the display of partner preference in ovariectomized, estrogen- and progesterone-primed rats. Preference for a sexually vigorous male or an estrous female rat was determined in one of two conditions: unlimited physical access to the stimulus rats (Contact condition) or access that was limited to olfactory, auditory and visual cues (No-contact condition). Lesions of the mPOA reduced the male preference, social preference, and arena crossings, independent of test condition. However, the reduction in male preference following mPOA lesions was most pronounced during tests with unlimited physical access. These results suggest that the mPOA may be involved in integrating somatosensory signals from coital stimulation with the motor responses associated with the appetitive aspects of female sexual behavior.

Hormonal status and test condition, but not sexual experience, modulate partner preference in female rats.

A series of experiments was conducted to determine the contributions of hormonal status, test condition, and sexual experience to the display of partner preference by female rats. Preference for a sexually active male rat over a sexually receptive female rat was assessed in independent groups of female rats tested in a condition limiting physical contact (No Contact) and a condition allowing for sexual interaction (Contact). Although hormonal status and test condition influenced the preference for a sexually active male, repeated testing and sexual experience had no effect. Experiment 1 demonstrated that independent of test condition, preference for the male is stronger in estrogen- and progesterone-primed rats than in rats receiving the vehicle. Moreover, independent of hormone condition, rats tested in the No Contact condition exhibit a stronger preference for the male than rats tested in the Contact condition, reflecting in part the active pacing of mating stimulation by sexually receptive rats tested in the Contact condition. Experiment 2 showed that the overall pattern of partner preference in proestrous and diestrous rats was similar to that observed in ovariectomized, estrogen- and progesterone-primed, and oil-treated rats, respectively. In Experiment 3, rats primed with estrogen alone did not exhibit a preference for the male even though fully receptive. Experiments 4 and 5 demonstrated that sexual experience does not affect the expression of preference for the male in estrogen- and progesterone-primed rats. The present findings demonstrate that the female rat's preference for the male is stable across repeated tests and is not affected by sexual experience. Our results also confirm that gonadal hormones influence the expression of a preference for a sexually active male versus a sexually receptive female and demonstrate that the magnitude of preference is modulated by test conditions.

Paced mating behavior in the female rat following lesions of three regions responsive to vaginocervical stimulation.

The present study examines the effects of ibotenic acid lesions of the medial amygdala, the bed nucleus of the stria terminalis and the medial preoptic area on the display of paced mating behavior in female rats. Lesions of either the medial amygdala or the bed nucleus of the stria terminalis have no effect on the display of paced mating behaviors in ovariectomized, hormone-primed rats. In contrast, lesions of the medial preoptic area significantly lengthen contact-return latencies following intromissions and ejaculations and increase withdrawal from the male following intromissions. The present study demonstrates that the medial amygdala and the bed nucleus of the stria terminalis are not involved in the behavioral responses accompanying paced mating behavior, whereas the medial preoptic area is a critical component of the neural circuit mediating paced mating behavior as well as other appetitive aspects of mating.

Amphetamine modulation of paced mating behavior.

The present study evaluated the effects of acute and repeated, intermittent amphetamine administration on paced mating behavior in ovariectomized (OVX) rats primed with estrogen and progesterone. In Experiment 1, female rats were tested for paced mating behavior following acute administration of amphetamine (1.0 mg/kg). Amphetamine increased the likelihood that a female would withdraw from a male following a mount or an intromission. Although this dose of amphetamine did not alter sexual receptivity or the latency to return to a male after sexual stimulation, locomotor activity was increased significantly. Experiment 2 evaluated the dose response characteristics of acute amphetamine (0.5, 1.0 and 2.0 mg/kg) administration on paced mating behavior. In agreement with Experiment 1, amphetamine at all doses increased the likelihood that a female would withdraw from a male following sexual stimulation. In Experiment 3, female rats were tested for partner preference (sexually active male vs. estrous female) following acute amphetamine administration. Amphetamine treatment augmented both social and sexual preferences. In Experiment 4, female rats were administered estrogen (20 microg/kg) and amphetamine (1.0 mg/kg) for 3 weeks and tested for paced mating behavior 1 and 4 weeks later, amphetamine free. Repeated intermittent exposure to amphetamine shortened the latency to return to a male after receiving a mount on the test conducted 1 week after the final drug injections. Collectively, these results suggest that the acute effects of amphetamine on paced mating behavior may reflect a reduction in social and sexual behaviors and an increase in locomotor activity, whereas the effects of repeated exposure may reflect a change in incentive motivation.

Behavioral and physiological responses to anabolic-androgenic steroids.

Anabolic-androgenic steroids (AAS) are synthetic derivatives of testosterone originally designed for therapeutic uses to provide enhanced anabolic potency with negligible androgenic effects. Although AAS continue to be used clinically today, the medical benefits of low therapeutic doses of AAS stand in sharp contrast to the potential health risks associated with the excessive doses self-administered not only by elite athletes and body builders, but by a growing number of recreational users, including adolescent boys and girls. The deleterious effects of AAS on peripheral organs and the incidence of altered behaviors in AAS abusers have been well documented in a number of excellent current reviews for clinical populations. However, a comparable synthesis of nonclinical studies has not been made. Our purpose in this review is to summarize the literature for animal models of the effects of supraphysiological doses of AAS (e.g. those that mimic human abuse regimes) on behaviors and on the neural circuitry for these behaviors. In particular, we have focused on studies in rodents that have examined how AAS alter aggression, sexual behaviors, anxiety, reward, learning, and locomotion and how AAS alter the expression and function of neurotransmitter systems and other signaling molecules that underlie these behaviors.