Sex with knockout models: behavioral studies of estrogen receptor alpha.

Estrogens are an important class of steroid hormones, having multiple targets, in the body and brain, and exerting ubiquitous effects on behavior. At present, two estrogen receptors (ERalpha and beta) have been cloned and sequenced in mammals. In the brain these receptors are regionally specific, but both have widespread distributions, which are largely non-overlapping. Given the newly emerging complexities of estrogen's mechanisms of action it is important to distinguish which pathways are involved in modifying which behaviors. We use a knockout mouse, lacking functional copies of the estrogen receptor alpha (ERalpha) gene, to study the mechanisms by which estrogens mediate behaviors. There are pronounced ramifications of ERalpha gene disruption on behavior. First, female ERalpha knockout (ERalphaKO) mice do not display normal feminine sexual behavior. Second, treatment of adult mice with androgens promotes masculine sexual behavior in both sexes. However, male-typical sexual behavior is severely compromised in male and female ERalphaKOs. Third, male ERalphaKOs do not exhibit the same social preferences for female mice as do wildtype (WT) littermates. Thus, the ERalpha is essential for normal expression of sexual behaviors. In addition, gonadectomized ERalphaKO and WT mice rapidly learn to escape from the Morris water maze. Exogenous estrogen treatment prevents WT females from learning this task, yet, has no effect in ERalphaKO mice, suggesting that estrogens effects on learning in adult females involves the ERalpha. Based on these data we hypothesize that ERalpha mediates many of the effects of estrogen on sexual behavior, learning, and memory.     

ERalpha, but not ERbeta, mediates the expression of sexual behavior in the female rat

Estrogen has well known effects on sexual behavior, however the role of the estrogen receptors (ER) alpha and beta on sexual behavior remains to be fully determined. This study investigated the individual and co-operative involvement of ERalpha and beta on sexual behaviors in the adult female rat. Subtype selective ER agonists, propyl-pyrazole triol (PPT; ERalpha agonist) and diarylpropionitrile (DPN; ERbeta agonist) were utilized to examine each receptor subtype's contribution, individual and co-operative, for both receptive (lordosis) and proceptive (hopping/darting, 'ear wiggling') female sexual behaviors. Ovariectomized female rats received subcutaneous injections of either: sesame oil (OIL), dimethylsulfoxide (DMSO), estradiol benzoate (EB; 10 microg/0.1 ml OIL), one of three doses of the ERalpha agonist PPT (1.25mg, 2.5mg or 5.0mg/0.1 ml DMSO), one of three doses of the ERbeta agonist DPN (1.25mg, 2.5mg or 5.0mg/0.1 ml DMSO) or a combination dose of PPT and DPN (2.5mg PPT+2.5mg DPN/0.1 ml DMSO) for two consecutive days, 48 and 24h prior to testing followed by a progesterone injection (500 microg/0.1 ml OIL) 4h prior to testing in order to elicit sexual behavior. The ERalpha agonist PPT, but not the ERbeta agonist DPN, elicited both proceptive and receptive behavior. PPT at doses of 2.5 and 5.0mg significantly elicited lordosis and proceptive behavior ('ear wiggling', hopping and darting). Intriguingly, the administration of both agonists together at the 2.5mg dose resulted in reduced levels of proceptivity and receptivity, suggesting that ERbeta modulates ERalpha's ability to elicit receptive and proceptive sexual behavior.     

Effects of nucleus accumbens lesion on female rat sexual receptivity and proceptivity in a partner preference paradigm

A partner preference paradigm, stud male vs estrous female, was used to study sexual behavior. Ovariectomized Wistar rats received bilateral electrolytic (n = 33) or sham (n = 16) lesion of the nucleus accumbens. Animals were tested in two different experimental situations, either with stimulus animals tethered (test with possibility of mating) or with stimulus animals behind a wire mesh (test without possibility of mating). Each test was carried out once prior to surgery and twice postoperatively following priming with estradiol benzoate and progesterone. Lordosis, rejection behaviors and soliciting patterns were scored in tests with tethered stimulus animals. The tendency of the experimental female to approach and remain in the vicinity of each stimulus animal was quantified to study the partner preference. Nucleus accumbens lesion increased the number of rejection responses to male mount attempts without modifying either receptivity estimated by lordosis reflex or soliciting behaviors. Control females showed a statistically significant preference for the male throughout the experiments. Lesioned females exhibit a preference for the male only in the test without sexual interaction possibility. This preference disappeared in the test with possibility of mating and it was even reversed when the male showed copulatory activity. These results, together with the absence of changes in lordosis behavior and soliciting activity, suggest that both the inversion in preference and the higher levels of mount rejections found in the lesioned animals are more likely attributable to hyperreactivity to the copulatory stimulus rather than an alteration in the female's rat sexual motivation.     

Preoptic lesions increase the display of lordosis by male rats

Male rats do not normally show feminine patterns of sexual behavior even when injected with the ovarian hormones estrogen and progesterone. We find that brain lesions which damage the preoptic-anterior hypothalamic continuum augment the display of lordosis in hormone-treated male rats. The most effectively feminizing brain lesions are ones which bilaterally destroy a substantial portion of the medial preoptic area encompassing the sexually dimorphic nucleus of the preoptic area (SDN-POA). Males with particularly large preoptic lesions are receptive following estrogen treatment and show a progesterone facilitation of receptivity. In this respect, they cannot be behaviorally distinguished from females. Thus, axons originating in and/or passing through the preoptic area apparently inhibit the display of feminine sexual behaviors in males. Preoptic development and lordosis are each predictably affected by perinatal stimulation by testicular hormones, and hormone-stimulated preoptic development may form the neurological basis for some of the defeminizing effects of perinatal hormonal exposure. Our results raise the possibility that the site of this behavioral defeminization is the SDN-POA.     

Female sexual behaviors in male rats with dorsal raphe nucleus lesions: Treatment with p-chlorophenylalanine

The effects of the serotonin-synthesis inhibitor, p-chlorophenylalanine (PCPA) on female sexual behaviors were examined in male rats with or without lesions (DRL) of the dorsal raphe nucleus, which contains a large number of serotonergic cell bodies. Estrogen-primed castrated males without brain surgery (control) showed extremely low levels of lordosis compared with females. On the other hand, DRL males displayed lordosis response more frequently than control males, but the lordosis quotient (LQ) in this group was lower than that in females. As well as DRL males, all PCPA-treated males showed lordosis, the mean LQ being comparable to the DRL group. Thus, the destruction of the dorsal raphe nucleus or the deprivation of serotonin by PCPA treatment facilitates manifestation of lordosis behavior in male rats. However, synergistic effect of DRL and PCPA treatments on female sexual behaviors have not been observed. The mean LQ in PCPA-treated male rats with DRL was almost the same as in DRL males or PCPA-treated males. These results suggest that the possible site of action of PCPA in regulating female sexual behavior in male rats is the serotonergic neurons in the dorsal raphe nucleus. Furthermore, the lordosis-facilitating effect of DRL is due to destruction of the serotonergic cell bodies in the dorsal raphe nucleus.     

Experimental assessments of behavioral and anatomical components of female chacma baboon (Papio ursinus) sexual attractiveness

Two experiments were carried out in order to separate the effects of the behavioral and the anatomical components of female sexual attractiveness in baboons. In the first experiment, two ovariectomized females were given increasing dosages of exogenous estradiol benzoate, and data were collected on their behavior as well as on male sexual arousal as assessed by the occurrence of coagulated seminal emissions of the type that result from masturbation. Male sexual arousal levels increased positively with the females' perineal swellings and appeared relatively unrelated to the females' behavior. In the second experiment, data were collected from five different groups of males in a systematic fashion across the period of a full menstrual cycle for their varied stimulus female partners. Behavior observations were carried out on the five different stimulus females, and ratings also were made of fluctuations in their perineal tumescence. The results of this experiment make it clear that male sexual arousal fluctuated across the study cycles in a fashion very similar to that of the perineal swellings of the stimulus females. Behavior was shown during all phases of the cycle, and, although it may have had a positive influence on male arousal during the follicular phase of the female's cycle, it had no apparent effect during the luteal phase. These results taken together establish for the first time that an estrogen stimulated female perineum per se has the potential to sexually arouse the male baboon. In addition, the data suggest that, although behavior may exert a positive effect on male arousal, in the absence of an estrogen stimulated perineum it is not sufficient to arouse males to masturbate. Finally the results establish that, although intact females appear to show behavior increases which may be arousing to males around the time of ovulation, these behaviors do not have to be shown by a female in order to arouse a male, provided she is swollen.     

A clinical investigation of the possible effects of long-term habituation of sexual arousal in assisted covert sensitization

Behavior principles are naturally implicated in most of the behavior therapy techniques used in the treatment of sexual dysfunctions and sexual disorders. Hypotheses differ, however, in the extent to which behavior therapy procedures emphasize the classical conditioning of sexual arousal (e.g. penile responses) or the operant conditioning of deviant sexual behavior patterns, or other often ignored behavior principles such as habituation. The purpose of the present case study is to investigate the possibility that at least part of the clinical effects of a widely used behavior therapy technique for sexual deviations, covert sensitization, may directly involve habituation processes. Results of the assisted covert sensitization case study indicate that, while expected decrements in sexual arousal to sexually inappropriate stimuli were observed, decrements in sexual arousal and self-report were also observed for sexually appropriate stimuli which received no aversive consequences, only repeated stimulus presentation over time. It is argued that habituation processes need further empirical investigation as a potential behavioral mechanism in the beneficial clinical effects usually noted in covert sensitization procedures.     

A review and reevaluation of the role of serotonin in the modulation of lordosis behavior in the female rat.

The role of serotonin (5-HT) in the modulation of sexual receptivity (lordosis) in the female rat is reviewed and reevaluated. The effects on lordosis of drug treatments that decrease or increase the activity and availability of central 5-HT are first discussed, and this is followed by an evaluation of the effects of drugs that act directly at 5-HT receptors. In order to shed light on the physiological significance of effects of serotonergic drugs on lordosis, there is also a review of what is known of changes in levels of serotonergic activity and densities of 5-HT receptors in the female rat brain that take place through the estrous cycle and in response to administration of behaviorally effective doses of gonadal steroids. Serotonin has generally been thought to have a tonic, inhibitory effect on lordosis. However, it is concluded that 5-HT can either inhibit or facilitate lordosis depending on which subtypes of central 5-HT receptors become activated. Because of a lack of consistent or compelling evidence of effects of ovarian hormones on serotonergic activity or 5-HT receptors in critical areas of the brain, it is stated that there is at present no basis to conclude that the effects of pharmacological manipulations of serotonergic activity on lordosis reflect an important, physiological role of 5-HT in the modulation of lordosis behavior in the female rat.     

Lesions of the preoptic area facilitate lordosis behavior in male and female guinea pigs

Male and female guinea pigs received radiofrequency lesions in the medial preoptic area (MPOA). Animals were gonadectomized, treated with estrogen and progesterone, and tested for the occurrence of the lordosis response to manual stimulation. Females with MPOA lesions exhibited enhanced lordosis behavior, shorter latencies to heat, longer duration of heat and longer maximum lordosis duration than sham control females. In males with MPOA lesions, the lordosis response could be elicited by manual stimulation, in contrast to no response in the sham control males. Furthermore, MPOA-lesioned males were insensitive to the inhibitory effects of progesterone on lordosis behavior, while MPOA-lesioned females were as sensitive as sham controls to the inhibitory effects of progesterone. The results suggest that a neural mechanism resides within the MPOA which inhibits the occurrence of lordosis behavior in both male and female guinea pigs and which is not involved in a sexual dimorphism in responsiveness to progesterone.     

Double oestrogen receptor alpha and beta knockout mice reveal differences in neural oestrogen-mediated progestin receptor induction and female sexual behaviour

To test the hypothesis that oestrogen receptor alpha (ERalpha) and ERbeta act together to mediate the actions of oestrogen in the ventromedial hypothalamus (VMH), we used mice with single or double knockout mutations of the ERalpha and ERbeta genes. Ovariectomized mice were implanted with 17beta-oestradiol and killed 5 days later. Oestradiol treatment promoted progestin receptor (PR)-immunoreactivity (-ir) in the VMH of all genotypes, but was maximal in brains of wild-type and ERbetaKO females. Analysis of specific VMH subregions revealed that PR-ir induction was limited to the caudal VMH in ERalphaKO and ERalphabetaKO mice. In the rostral VMH, oestradiol only induced PR-ir in wild-type and ERbetaKO mice, and the number of PR-ir neurones in this region was greater in ERbetaKO than wild-type females. Next, we tested the ability of a dopamine agonist and progesterone to facilitate sexual behaviour in females lacking functional ERalpha, ERbeta, or both receptors. Ovariectomized mice were implanted with oestradiol, and tested for sexual behaviour three times after administration of the dopamine agonist, apomorphine, followed by two tests concurrent with progesterone treatment and a final test with just apomorphine treatment. ERalphaKO and ERalphabetaKO females failed to display lordosis under any testing conditions, while ERbetaKO females exhibited lordosis behaviour equal to that of wild-type females. Our data show that a subpopulation of PR-ir neurones is induced by oestradiol in the caudal VMH of female mice lacking both ERalpha and ERbeta genes. We hypothesize that this action of oestradiol is either mediated by a novel ER or by the mutant portion of the AF2 subregion of the ERalpha gene present in ERalphaKO brain. However, despite the presence of PR in VMH, females lacking a functional ERalpha gene do not display sexual behaviour, via either ligand-dependent or -independent activation.     

Gender identity change in transsexuals. Follow-up and replications.

An apparently successful change in gender identity of an adolescent transsexual was reported several years ago. Gender-specific motor behavior, appropriate sex role social behavior, cognitive sexual activity, and finally sexual arousal patterns were defined, measured, and sequentially modified. In this report, a 6 1/2-year follow-up is presented and the application of a similar therapeutic package to two additional cases is described. In the first case, sweeping changes in gender identity, sex-role behavior, and sexual arousal patterns were observed. In the second of these cases, rigid feminine gender was given up, but the patient chose to retain homosexual arousal. Social adaptation was satisfactory. These findings and other recent developments point to the possibility of psychosocial intervention as an alternative to surgery in the treatment of transsexualism.     

Evidence that the effect of melanocortins on female sexual behavior in preoptic area is mediated by the MC3 receptor; Participation of nitric oxide

alpha-MSH is involved in reproductive processes and can regulate the expression of lordosis, an important component of female reproductive behavior in rats and many other species. In this study, we investigated the effects of MSH peptides on lordosis behavior when injected in medial preoptic area (POA) of ovariectomised rats primed with estradiol. The results show an increase in lordotic activity after bilateral administration of alpha-MSH and gamma-MSH. Interestingly, the treatment with the MC4 receptor antagonist HS014 did not block the stimulatory effect of alpha-MSH. Moreover, the injection of HS014 did not itself modify the lordosis quotient. Nitric oxide has been suggested to play a crucial role in the regulation of lordosis behavior via stimulation of guanylyl cyclase to synthesize cGMP. In order to determine the participation of NO in the effect of the melanocortins, another group of rats were treated with L-NAME, an inhibitor of NOS, alone or 15 min before the injection of alpha-MSH or gamma-MSH. The injection of L-NAME into the POA of E-primed rats 15 min before the test for sexual receptivity did not modify significantly the lordosis quotient at the two doses examined. The treatment with L-NAME at the lowest dose completely abolished the stimulatory effect of alpha-MSH and gamma-MSH on sexual behavior. The results indicate that the effects of MSH peptides on female sexual behavior in this area are mediated through specific MC receptor, that could be the MC3 receptor and that NO mediates the melanocortins effects.     

Self-related processing in the sexual domain: a parametric event-related fMRI study reveals neural activity in ventral cortical midline structures

Self-related processing, reflecting the evaluation of environmental signals with regard to personal relevance, is fundamental for decision-making and subsequent behavioral responses. While self-related processing has already been investigated in several domains, one important domain, the sexual domain, has been spared so far. Recent imaging studies suggest that self-related processing in different domains involves common regions in medial orbitofrontal and prefrontal cortex, the so-called ventral cortical midline structures (CMS). However, the same regions have also been implicated in sexual arousal, especially with regard to emotional processing in sexual arousal. Therefore it remains unclear whether this involvement of ventral cortical midline regions reflects emotional processing in sexual arousal or associated self-relatedness. We here report data from a parametric event-related fMRI study that investigated the neural correlates of self-related processing in sexual arousal, using erotic pictures from the International Affective Picture System. It was found that self-related activity associated with sexual arousal showed neural activity in ventral CMS regions such as the venteromedial prefrontal cortex (VMPFC) and the perigenual anterior cingulate cortex (PACC), while self-related activity not associated with sexual arousal showed neural activity in the dorsomedial prefrontal cortex (DMPFC). Our study indicates that self-relatedness may be considered a crucial component in sexual arousal that is mediated by neural activity in ventral cortical midline structures.     

Interactions between estrogen and progesterone in neural tissues that mediate sexual behavior of guinea pigs

(1) Interactions of estrogen and progesterone with each other and with the neural tissues that regulate sexual behavior in female guinea pigs were studied. (2) Long-acting preparations of estradiol-17β (E₂) were more effective in facilitation of lordosis than other estrogens. (3) A major behavior-facilitating site of action of E₂ is in the medial basal hypothalamus. (4) E₂ is selectively taken up by a saturable receptor system in fractions of hypothalamic tissue. (5) Although early effects of E₂ on neural tissues that mediate lordosis can be mimicked by certain anti-estrogens, anti-estrogens do not mimic long-term effects of E₂ that are required for optimum expression of lordosis behavior. (6) Progesterone (P) is required in extremely small quantities for facilitation of lordosis behavior in estrogen-primed female guinea pigs. (7) Facilitation of sexual behavior is a short-term effect of P that is mediated by the medial basal hypothalamus. (8) P inhibits the expression of lordosis behavior via a mechanism that is represented in the midbrain. (9) This inhibitory action of P has a longer latency than the facilitatory action of P. (10) Prolonged residence of E₂ in the hypothalamus temporarily favors the expression of short-term facilitatory actions of P on lordosis. (11) The long-term lordosis-antagonizing effects of P are not due to inhibition of E₂ uptake in the hypothalamus. (12) Inhibitory effects of P on lordosis may not depend on neural cells that have the ability to concentrate E₂ in their nuclei.     

The role of melanocortin receptors in sexual behavior in female rats

Earlier data have indicated that alpha-MSH may play a role for sexual behavior in rats. In this study we investigated the effects of MSH peptides on sexual receptivity in ovariectomized-adrenalectomized female rats, pre-treated with benzoate of estradiol, in presence of vigorous male rats. The results show that alpha-MSH significantly increases lordosis behavior in female rats after injections into the ventromedial nucleus. Interestingly, we have for the first time shown that gamma-MSH also causes significant increase in lordosis in female rats. Furthermore, we show that HS014, an antagonist for the central MC receptors, in dose dependent manner blocks the effect of alpha-MSH on lordosis. The results indicate that the effects of MSH peptides on female sexual behaviour are mediated through a specific MC receptor, which could be the MC3 receptor.     

Self-related processing in the sexual domain: A parametric event-related fMRI study reveals neural activity in ventral cortical midline structures

Abstract

Self-related processing, reflecting the evaluation of environmental signals with regard to personal relevance, is fundamental for decision-making and subsequent behavioral responses. While self-related processing has already been investigated in several domains, one important domain, the sexual domain, has been spared so far.

Recent imaging studies suggest that self-related processing in different domains involves common regions in medial orbitofrontal and prefrontal cortex, the so-called ventral cortical midline structures (CMS). However, the same regions have also been implicated in sexual arousal, especially with regard to emotional processing in sexual arousal. Therefore it remains unclear whether this involvement of ventral cortical midline regions reflects emotional processing in sexual arousal or associated self-relatedness. We here report data from a parametric event-related fMRI study that investigated the neural correlates of self-related processing in sexual arousal, using erotic pictures from the International Affective Picture System. It was found that self-related activity associated with sexual arousal showed neural activity in ventral CMS regions such as the venteromedial prefrontal cortex (VMPFC) and the perigenual anterior cingulate cortex (PACC), while self-related activity not associated with sexual arousal showed neural activity in the dorsomedial prefrontal cortex (DMPFC). Our study indicates that self-relatedness may be considered a crucial component in sexual arousal that is mediated by neural activity in ventral cortical midline structures.     

Site-specific inhibition of receptivity by intracranial anisomycin in hamsters

The ventromedial nucleus of the hypothalamus (VMH) has been implicated in the mediation of the hormonal control of female rodent sexual behavior. However, in hamsters, progesterone (P) has been found to have effects on sexual receptivity in other diencephalic and mesencephalic sites as well. Progesterone is thought to exert its behavioral effects by altering protein synthesis in CNS target neurons. We tested the effects of 30 gauge implants of the protein synthesis inhibitor anisomycin in the preoptic area (POA), VMH, and ventral mesencephalon (VMES) 30 minutes before 500 micrograms P SC, on the facilitation of lordosis in ovariectomized estrogen-primed female hamsters. The same animals were tested one week later with estrogen and progesterone treatment but without anisomycin. Anisomycin reduced sexual receptivity (lordosis) when placed in the VMH or VMES, but not when delivered to the POA. The results confirm the importance of the VMH in the mediation of progesterone facilitation of female sexual behavior, but also provide evidence that ventral midbrain structures may play a role in female sexual receptivity in hamsters. These two structures may be important for different aspects of lordosis. Progesterone effects in both sites appear to be protein synthesis dependent.     

Female sexual behavior in the golden hamster following kainic acid lesions in the lateral septal area

Patterns of female sexual behavior in the hamster are qualitatively different from that observed in the rat. Hamsters demonstrate tonic immobility and a sustained lordotic response that can last for several minutes following minimal stimulation from the male while in rats, the lordotic response is highly dependent upon continued stimulation from the male and lasts for only a few seconds following a mount or intromission. Also, the hamster requires both estrogen and progesterone priming in order to display female sexual behavior but the rat can show lordotic behavior following treatment with estrogen alone. Electrolytic lesions in the lateral septal area of female rats produce a facilitation in female sexual behavior and increase behavioral sensitivity to estrogen. However, lesions in the lateral septal area of rats produced by the neurotoxin kainic acid (KA) result in deficits in female sexual behavior following estrogen and progesterone treatment. Given these species differences in sexual behavior, we have examined the effects of KA lesions in the lateral septal area of hamsters on female sexual behavior. Adult female golden hamsters were ovariectomized and tested for female sexual behavior prior to receiving brain surgery. For all behavior tests, animals were injected SC with 60 micrograms of estradiol benzoate/kg per day for two days and 0.5 mg of progesterone three hours prior to the behavior test on day three. Behavior tests consisted of placing the test animal in a male's home cage for 10 minutes and recording the latency to the first lordosis, length of the longest single bout of lordosis and total lordosis duration.(ABSTRACT TRUNCATED AT 250 WORDS)