Hormone-dependent aggression in female rats: testosterone implants attenuate the decline in aggression following ovariectomy

Female rats were individually housed with a sterile male for a 4- to 5-week period. Each female was then tested for aggression toward an unfamiliar female intruder at weekly intervals. Those females that displayed a high level of aggression on each of three weekly tests were ovariectomized and given subcutaneous implants of testosterone-filled tubes, ovariectomized and given subcutaneous implants of empty tubes, or sham-ovariectomized and implanted with empty tubes. These implants should produce a serum testosterone concentration of about 0.6 ng/ml, compared to 0.17 ng/ml in intact females. Beginning 1 week postoperatively, the aggression of each female was tested weekly for 4 weeks. Ovariectomized females with testosterone implants displayed a level of aggression significantly higher than that of ovariectomized females with empty implants on 3 of 4 weekly tests. The level of aggression by females with testosterone implants was not significantly different from that of sham-ovariectomized females on the first postoperative test. Additional observations showed that testosterone implants did not produce an increase in aggression in females whose preoperative level of aggression was low. Further, Silastic implants containing estrogen (1 to 2 mm long) sufficient to maintain a serum estrogen level of 20 to 30 pg/ml also attenuated the decline of aggression following ovariectomy. These results suggest that testosterone and estrogen may both contribute to the biological substrate of hormone-dependent aggression in female rats.     

Activation of POMC neurons during general arousal but not sexual behavior in male rats

Exogenous opioids influence male rat sexual behavior, suggesting that endogenous opioid peptides are released during mating. Supporting this hypothesis, the authors recently showed that mating induced activation of mu opioid receptors. However, it is unknown which ligand(s) is acting on these receptors during mating. The current set of experiments tested the hypothesis that beta-endorphin-producing neurons, that is, proopiomelanocortin (POMC) neurons, are activated during sexual behavior. Mating-induced activation of POMC neurons was investigated during either the dark phase or the light phase, following different components of male rat sexual behavior or following control manipulations that resulted in general arousal. Results show activation of POMC neurons in the mediobasal hypothalamus following general arousal but not specifically related to sexual behavior per se. In addition, mating did not activate the subpopulation of POMC neurons that project to the medial preoptic nucleus. These results suggest that it is unlikely that POMC neurons contribute to the action of endogenous opioids in the brain area during sexual behavior but instead may contribute to the change in arousal state essential for the expression of sexual behavior.     

Cohabitation with a female activates testosterone-dependent social aggression in male rats independently of changes in serum testosterone concentration

Male hooded rats (350 to 450 g) were sham-castrated, castrated and implanted with testosterone-filled, or castrated and implanted with empty Silastic tubes. Twenty-four hours postoperatively the animals in each group were housed with a female or a male similar in size to the female. Beginning one week following surgery and continuing for three weeks thereafter, the female or male cagemate was removed once each week while a 15-min test of aggression toward an unfamiliar male intruder was conducted. During the aggression tests, lateral attacks, lunge attacks, bites, on-top, and piloerection were recorded. At the first aggression test, males housed with females were significantly more aggressive than their counterparts housed with males. In contrast, different testosterone regimes did not consistently influence the initial activation of intermale social aggression. At the second and third tests, males with testicular testosterone or a replacement were significantly more aggressive than their castrated controls on most measures but males housed with females continued to be more aggressive than the comparable group housed with males. These results suggest that normal fluctuations in serum testosterone concentration associated with sexual interaction are not necessary for the initial activation of intermale social aggression. Both repeated exposure to unfamiliar males as well as cohabitation with a female are effective stimuli for activation of testosterone-dependent social aggression.     

Parachlorophenylalanine-induced serotonin depletion increases offensive but not defensive aggression in male rats

Cerebral 5-HT depletion has been shown to facilitate elicitation of various kinds of aggressive behavior in rats. The question as to whether both offensive and defensive aggressive reactions are affected to the same extent was examined in a resident-intruder paradigm where an ethological analysis of the two animals allows an evaluation of non-social activities as well as agonistic interactions, including both offense and defense. PCPA (375 mg/kg IP) was administered either to the resident or the intruder and the interactions with an untreated conspecific were recorded in the resident's home cage for an 8 min period three days after injection when 5-HT was maximally reduced. PCPA treatment increased the occurrence of social approach and offensive postures in resident rats, whereas their untreated partners displayed more defensive reactions. When intruders were injected, only non-significant increases in approach and offense were observed. In no case did PCPA affect occurrence of defensive postures in the injected animals. These results confirm that serotonin plays a role in controlling offensive aggression but not defensive behavior.     

Naltrexone effects on male sexual behavior, corticosterone, and testosterone in stressed male rats

Chronic physical or psychological stress disrupts male reproductive function. Studies in our laboratory have shown that stress by immersion in cold water (ICW) and by electrical foot shocks (EFS) has inhibitory effects on male sexual behavior; these effects do not seem to be mediated by an increase in corticosterone, nor by a decrease in testosterone. On the other hand, it is known that endogenous opioids are released in the brain in response to these same stressors; consequently, they could be participating in the impairment of sexual behavior, as well as in the changes in corticosterone and testosterone caused by stress. The aim of this study was to analyze the effects of the opioid antagonist naltrexone (NTX) on male sexual behavior, corticosterone, and testosterone in both stressed sexually experienced and naive male rats. Sexually experienced adult male rats were assigned to one of the following groups (n = 10 each): 1) control group, males without sexual evaluation; 2) control group, rats injected ip with saline, non-stressed; 3) control group, rats injected with NTX (3 mg/kg) non-stressed; 4) rats injected ip with saline, and stressed by EFS; 5) rats injected ip with NTX (1.5 mg/kg) and stressed by EFS; 6) rats injected ip with saline and stressed by ICW; 7) rats injected ip with NTX (1.5 mg/kg) and stressed by ICW; 8) rats injected ip with NTX (3 mg/kg) and stressed by ICW. Naive males were assigned to the same control groups but only stressed by ICW and the NTX dose used was 3 mg/kg. Injections were given 30 min before stress sessions. Stress was applied on 20 consecutive days. Male sexual behavior was assessed 15 min after EFS or 30 min after ICW, on days 1, 4, 8, 12, 15, and 20. Trunk blood was collected at the end of the experiments on day 20 of stress. Corticosterone and testosterone were evaluated by HPLC.Mount, intromission and ejaculation latencies were longer in control saline naive males compared to control saline sexually experienced males on the first day. NTX administration to control naive males caused a decrease in mount, intromission, and ejaculation latencies, as well as an increase in ejaculatory frequency/30 min, compared to control-saline only on day 1. Stressed naive males showed higher mount, intromission and ejaculation latencies, compared to control and stressed sexually experienced males, as well as comparable increase in corticosterone and decrease in testosterone plasma levels. NTX administration before exposure to stress prevented the modifications caused by stress in sexual parameters. Sexual behavior in control sexually-active males injected with saline or NTX was not modified. Saline stressed males showed the previously reported alterations in sexual behavior, as well as an increase in corticosterone and a decrease in testosterone plasma levels. Stressed males injected with NTX before exposure to stress showed no alterations in male sexual behavior. NTX in control non-stressed males did not modify corticosterone plasma levels, but did cause a significant increase in plasma testosterone. The increase in corticosterone and the decrease in testosterone due to stress, were attenuated with the opioid antagonist, both in naive and sexually experienced males. Prevention of ICW stress effects was more effective with higher doses of NTX (3 mg/kg). These data suggest that endogenous opioids could be participating in the effects caused by stress on male sexual behavior, corticosterone, and testosterone.     

Dihydrotestosterone activates sexual behavior in adult male hamsters but not in juveniles

The effect of an androgenic metabolite of testosterone, dihydrotestosterone (DHT), on reproductive behavior and brain androgen receptor (AR) immunoreactivity was compared in juvenile and adult male Syrian hamsters. Prepubertal and adult animals were castrated and treated with 0, 500, or 1000 microg of DHT daily for 1 week and then tested for their ability to engage in mating behavior. The 1000-microg dose of DHT activated intromissions in adult but not prepubertal males. Brains were collected immediately after the behavioral test to investigate whether the lack of a behavioral response to DHT prior to puberty is associated with fewer AR-immunoreactive (AR-ir) cells in the forebrain nuclei that mediate male sexual behavior. In four of the five nuclei within the behavioral circuit that were examined, the number of AR-containing cells was similar in prepubertal and adult males treated with 1000 microg of DHT. Only in the anterior medial amygdala (MeA) was there a greater number of AR-ir cells in adults. These data indicate that (1) DHT does not activate components of male reproductive behavior prior to puberty and (2) the lack of behavioral responsiveness to DHT in prepubertal males is most likely not related to an overall reduction in ARs within the forebrain circuit that mediates mating behavior.     

Serum leptin correlates with serum uric acid but not serum testosterone in non-obese male adolescents

To identify the serum factors that affect circulating leptin levels, we measured the serum concentrations of leptin, testosterone (T), estradiol (E), serum alanine aminotransferase, total cholesterol and uric acid (UA) in healthy male adolescents (age, 18.3 +/- 0.1 years, n=96). We also measured body mass index (BMI), percent body fat and thickness of skin fold to assess the effect of body constitution on serum leptin level. Since serum concentration of leptin significantly correlated with BMI (r=0.820, p<0.001), we analyzed the relation-ship between leptin/BMI ratio (L/BMI) and serum parameters. Analysis of data of subjects with normal serum T level showed a significant inverse correlation between L/BMI and serum T levels (n=96, r=-0.294, p<0.005), but no such correlation was present among non-obese subjects (n=70) with BMI of +/-20% of normal (22 kg/m2). There was no correlation between L/BMI and serum E level. Serum UA level significantly correlated with L/BMI in both the test group (n=96, r=0.520, p<0.001) and non-obese subjects (r=0.369, p<0.005). Stepwise multiple regression analysis showed that UA independently and significantly influenced serum leptin levels in both the test and control groups. Our results demonstrate that T weakly influences serum leptin concentration, and that UA concentrations strongly influences serum leptin in healthy male adolescents independent of their obesity level.     

Relation of autogrooming to sexual behavior in male rats

Grooming and penile reflexes were studied in male rats that were restrained in supine position with the penile sheath retracted or were free to copulate with sexually receptive females. In Experiment 1 there was a reliable concordance in supine males between the tendency to groom and the tendency to display penile reflexes. In Experiment 2 we analyzed the sequential organization of grooming and genital events in supine tests. It was assumed that many or most episodes of ventral grooming would have been genital grooming had access to the genitalia not been prevented by restraint. Paw grooming tended to precede clusters of penile responses, whereas ventral grooming started after the onset of erections. Experiment 3 was an exploration of grooming in the context of copulation, rather than supine restraint. Males groomed their genitalia immediately after all intromissions and after all mounts that ended mount bouts. The duration of grooming was not affected by whether or not intromission occurred. Finally, in Experiment 4 we observed genital and nongenital grooming and recorded electromyographic (EMG) activity from the striated bulbospongiosus muscle (mBS) of the penis in freely moving rats. Consistently, mBS activity led to genital grooming with a short latency, whereas nongenital grooming rarely led to genital grooming, and EMG activity was not associated with nongenital grooming nor did it tend to follow after genital grooming was initiated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Neonatal testosterone administration, but not in utero contiguity to males, augments the display of male sexual behavior by testosterone-treated adult female mice

Male copulatory behavior of adult female mice given slow-release capsules of testosterone was examined in animals that developed in utero contiguous to two males (mFm) or to two females (fFf). Other females of unspecified uterine position which were injected with testosterone propionate on the day of birth as well as intact males also were examined. mFm and fFf females did not differ on any measure; latency to the first mount, number of mount bouts, number of mount bouts with genital thrusting. The perinatally androgenized females exhibited more mount bouts and more bouts accompanied by genital thrusting than did mFm and fFf subjects. The former also displayed more mount bouts with thrusting on the second pair of tests than males. Lastly, a greater proportion of perinatally androgenized females than mFm or fFf animals displayed male sexual behavior two weeks following removal of the testosterone-containing capsule.     

Activation of sexual reflexes of male rats by dihydrotestosterone but not estrogen

Previous studies have shown that withdrawal and administration of testosterone propionate (TP) has a quantitative influence on sexual reflexes which parallels changes in copulatory activity following castration and administration of TP. The present study involving castrated spinal male rats explored further this parallel, focusing on the effects on sexual reflexes of the administration of dihydrotestosterone propionate (DHTP) and estradiol benzoate (EB), both of which can activate sexual behavior in spinally intact castrated male rats, but only if given in very large doses for a prolonged period of time. A parallel effect on reflexes and behavior was not found inasmuch as DHTP activated sexual reflexes at a dose (200 microgram daily) considerably below that needed to activate behavior, and EB did not appreciably activate reflexes, even after prolonged treatment at levels (100-200 microgram) higher than necessary to activate behavior. The results, with EB in particular, point out that the display of intromissive and ejaculatory patterns in rats may not involve spinal neural mechanisms that are customarily associated with these behavioral patterns.     

Stress and sexual behavior in male rats

The relationship between sexual behavior and stress in male rats was investigated. Stress induction by psychophysical immobilization fo 3 hours along three days caused significant alterations of the different parameters which make up the sexual behavior of male rats. An extension of the stressing situation from 3 to 6 hours under the same experimental conditions produced very similar sexual disturbances. Our experimental results suggest that effects of acute stress on sexuality may be independent of time.     

Hippocampal long-term potentiation is reduced in mature compared to young male rats but not in female rats

Aging is associated with a decline in cognitive function which could be due to reduced synaptic plasticity. Hippocampal long-term potentiation (LTP) is an activity-dependent form of increased transmission efficacy at synapses that is considered the basis for some forms of learning and memory. We studied the N-methyl-d-aspartic acid (NMDA) receptor-dependent LTP in the CA1 region of hippocampus in young (2 months) and mature (8 months) male and female rats. We have found that in young male rats the tetanus increased the magnitude of excitatory post-synaptic potentials to 204+/-10% of basal while in mature male rats the magnitude of the LTP was significantly lower reaching only 153+/-11% of basal. This decrease did not occur in female rats. Similar changes occurred in the content of the NMDA receptor subunits NR1 and NR2A in hippocampus. The amount of both subunits was reduced significantly (15-16%) in hippocampus of 8-month-old compared with 2-month-old male rats. This decrease was not observed in female rats. Moreover, there is a significant correlation between the content of NR1 subunit and the magnitude of the potentiation. These data suggest that some of the neurobiological changes induced in hippocampus by aging are different in males and females.     

Hormone-dependent aggression in the female rat: testosterone plus estradiol implants prevent the decline in aggression following ovariectomy

Female rats were individually housed with a sterile male for the duration of the experiment. Beginning 7 to 10 weeks after the start of cohabitation, each female was tested for aggression toward an unfamiliar female at weekly intervals for 3 weeks. Females that displayed consistent and substantial aggression were given one of the following treatments: ovariectomy followed by both testosterone and estradiol implants, ovariectomy followed by 2 empty implants, or sham ovariectomy followed by 2 empty implants. The implants were subcutaneously placed hormone-filled Silastic capsules. They were expected to produce a serum testosterone concentration of 0.5 ng/ml and an estradiol concentration of 15 pg/ml. Postoperatively, the aggression of each female continued to be assessed on a weekly basis for 3 weeks. Ovariectomized females with hormone implants displayed a level of aggression postoperatively similar to that of sham-ovariectomized females and significantly greater than that of ovariectomized females with empty implants. These results, together with others, suggest that estradiol and testosterone act together to form the hormonal foundation of hormone-dependent aggression by females cohabiting with a sterile male.     

Age-related deficits in brain androgen binding and metabolism, testosterone, and sexual behavior of male rats

Brain androgen binding and metabolism, serum testosterone (T), and sexual behavior were measured in old and young male Fischer 344 rats. After completion of sexual behavior tests, blood was collected for T assay and brains were removed for simultaneous measurements of cytosolic (ARc) and nuclear (ARn) androgen receptors and aromatase activity (AA) in the preoptic area (POA), hypothalamus (HYP) and amygdala (AMG). In Experiment 1, old and young intact males were examined. None of the old males ejaculated in any of the tests of sexual behavior whereas all of the young males ejaculated. The old males had lower levels of serum T, lower levels of ARn in the POA and HYP and lower levels of AA in the POA. The ARc levels of the old and young males did not differ. Experiment 2 was designed to determine if the deficits in brain androgen binding and metabolism were due to low levels of T. Old and young T-treated gonadectomized (GX-T) males and young intact (I) males were examined. T levels were comparable in the young and old GX-T males and were higher in each of these groups than in the young I males. In sexual behavior tests, all of the young but only 25% of the old GX-T males ejaculated. Although ARn levels in the old GX-T males were lower than in the young GX-T males, they were comparable to the young I male levels. No age-related differences in T induction of AA were observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Infradian fluctuations in serum testosterone levels in male laboratory rats

The dynamics of serum testosterone was studied in laboratory male rats kept separately from females. Infradian rhythm of changes in testosterone level equal to 4 days and ultradian (within 24 h) periods equal to 160 and 480 min were detected. The maximum serum level of testosterone in male rats was synchronized with the greatest number of estrous females.     

Prostaglandin E2 accelerates sexual behavior in male rats

Sexual behavior in male rats is accompanied by an increase in body temperature of 18 degrees C. It has been suggested that this increase may be, at least in part, a febrile response mediated by the endogenous central release of prostaglandins of the E series (PGE). This putative release of PGE could also affect the expression of sexual behavior, a possibility that was tested in the present experiment. PGE2 was infused into the cerebral aqueduct and sexual behavior and hypothalamic temperature were monitored. PGE2 infusion raised hypothalamic temperature and decreased the postejaculatory interval and ejaculation latency. The exact cause of this acceleration of sexual behavior cannot as yet be determined.     

Self-injurious behavior in male rhesus macaques does not reflect externally directed aggression

Self-injurious behaviors (SIB), such as self-biting and self-wounding, have been observed in a small percentage of captive nonhuman primates. Because rhesus monkeys that exhibit SIB also tend to be more aggressive, it was hypothesized that SIB is related to externally directed aggression and is associated with contexts in which physical contact between participants is prevented. The purpose of this study was to test the hypothesized relationship between SIB and outward aggression. Subjects were first presented with videotapes of conspecifics, scenery and a blank screen, and their behavior was recorded. Levels of salivary cortisol, an indicator of stress, were also measured before and after presentation of the videos. Although aggression increased when subjects viewed tapes containing conspecifics, neither cortisol levels nor self-biting behavior varied as a function of tape content. The subjects were then placed in two additional test situations: an empty room and the same room containing an unfamiliar conspecific. Aggression was significantly higher in the stranger condition compared to the empty room condition. The two situations yielded parallel increases in cortisol, suggesting that being alone was just as stressful as being paired with an unfamiliar conspecific. Self-biting rates were also similar in these two conditions. Thus, contrary to our prediction, increases in aggression did not correlate with increases in SIB. These results suggest that under similarly stressful conditions, SIB and externally directed aggression are unrelated.     

Coexpression of sexual behavior and maternal aggression: the ambivalence of sexually active mother rats toward male intruders

The sexually active female rat solicits the male to approach for copulation, while the maternal dam displays aggression to expel him from the nest, suggesting that both behaviors are mutually exclusive. However, the rat has a postpartum estrus during which she is sexual and maternally motivated. Can she perceive the male as attractive and aversive, soliciting and attacking him at the same time? This study shows that postpartum estrous females exhibit a merge of sexual and maternal aggressive responses toward male intruders in the home cage. The concurrent expression of these behaviors did not affect their intensities, although the stimulation of maternal behavior increased maternal aggression without modifying sexual solicitation. These results indicate that the postpartum estrous rat can optimally express two opposite and independently regulated motivations, and that the male can be perceived as an ambivalent stimulus.     

Evidence implicating aromatization of testosterone in the regulation of male ferret sexual behavior

We compared the effects of the aromatase inhibitor, 1,4,6-androstatriene-3,17-dione (ATD) and castration on the expression of mating behavior in adult male ferrets which were in breeding condition. Males implanted SC with Silastic capsules containing ATD displayed significantly less neck gripping, mounting and intromittive behavior than intact males which received empty capsules, although the ATD-induced reductions in behavior were not as large as those seen after castration. ATD had no effect on mating behavior in castrated males. As reported in another publication, brain aromatase activity was significantly reduced in both the intact and castrated males treated with ATD in the present study. Plasma estradiol (E2) levels were uniformly low in intact and castrated males, regardless of whether they received ATD or no steroid. As expected, plasma testosterone (T) levels were significantly lower in castrated than in intact males, and ATD treatment did not affect these values. These results suggest that E2 formed via the neural aromatization of T contributes to the activation of masculine sexual behavior in intact male ferrets in breeding condition.     

Sexual motivation is demasculinized, but not feminized, in prenatally stressed male rats

Sexual motivation and copulation in male rats are associated with dopamine release in the nucleus accumbens. Demasculinized copulatory behavior has been demonstrated in prenatally stressed adult male rats. We have previously reported that approximately 80% of prenatally stressed male rats do not exhibit copulation and that no significant changes in nucleus accumbens dopamine release are seen during exposure to estrous females. In the present study, we investigated whether prenatal stress affects sexual motivation in these animals as adults. Pregnant Wistar rats were subjected to immobilization stress for two hours daily from day 15-19 of gestation. The prenatally stressed male offspring at the age of 3 months were allowed contact with receptive female rats for a 30 min period per week for 10 weeks; then, between the age of 5 and 6 months, their sexual motivation and copulatory activity were measured. Sexual motivation was measured in terms of sexual partner preference. The number of visits and the duration of each visit to an estrous female (stimulus female) or to a sexually active male rat (stimulus male) were recorded. Compared with control males, prenatally stressed male rats showed a significantly lower number of visits and a shorter duration of each visit to stimulus females. Prenatally stressed males showed no preference for male or female stimulus rats in terms of the number of visits and the duration of each visit, whereas control rats showed a significantly higher number of visits and duration of visits to female stimulus rats than male stimulus rats. A significant decrease in copulatory activity was observed in the prenatally stressed male offspring compared with control male rats, with most of the prenatally stressed males failing to show copulation. In vivo microdialysis experiments were performed on the nucleus accumbens with concurrent observation of sexual behavior. The prenatally stressed rats that did not exhibit copulation showed no significant changes in nucleus accumbens dopamine release during exposure to a stimulus male behind a wire-mesh barrier and the amount of dopamine release remained at the basal levels during actual physical contact. These results, combined with those of our previous report, indicate that sexual motivation in prenatally stressed male rats is demasculinized, but not feminized.