Doris Zumpe

Intracerebral Infusion of an Aromatase Inhibitor, Sexual Behavior and Brain Estrogen Receptor-Like Immunoreactivity in Intact Male Rats

Copulatory behavior was studied in five groups of sexually experienced, gonadally intact male rats in which: (i) the nonsteroidal aromatase inhibitor Fadrozole (CIBA-Geigy CGS 16949A) was delivered bilaterally (0.756 microgram in 6.0 microliters saline in 24 h to each side) into the medial preoptic area (POM) together with normal saline given s.c. via osmostic minipumps (n = 10); (ii) normal saline was delivered bilaterally into POM together with the same dose of Fadrozole s.c. (n = 9); (iii) Fadrozole was delivered bilaterally into the lateral preoptic area together with normal saline given s.c. (n = 6); (iv) Fadrozole was delivered bilaterally into the cerebral cortex together with normal saline given s.c. (n = 6), and (v) unoperated controls (n = 14). Mounting and ejaculation were significantly decreased in rats receiving Fadrozole in POM compared with the behavior of rats in the other 4 groups. Few differences occurred between rats in the latter 4 groups, all of which continued to mate. The H222 and ER-715 anti-estrogen receptor (ER) antibodies were used to examine the distribution of ER immunoreactive (ERir) neurons in hypothalamic and limbic sites in gonadectomized controls and in some of the rats in groups i, ii and v. Since labeling of ERir neurons in rat brain with the H222 anti-ER antibody is reported to be inhibited by estrogen, it was used here to identify regions (with staining) where the aromatization of testosterone (T) into estradiol (E2) had been suppressed. Intense H222 ERir nuclear neuronal labeling was confined to the POM of males receiving Fadrozole in POM, and significantly more labeled neurons were found in the POM of these rats than in the POM of rats treated with saline in POM. In contrast, the ER-715 antibody, which is reported to stain neurons independently of hormonal status, labeled neuronal nuclei in hypothalamic and limbic regions of all groups, demonstrating the presence of ER. These findings show that conversion of T into E2 in the POM of the male rat is important for male rat copulatory behavior and that H222 ERir nuclear neuronal labeling can be used to identify the neurons in POM that were affected by Fadrozole.

Fos Induced by Mating or Noncontact Sociosexual Interaction Is Colocalized with Androgen Receptors in Neurons within the Forebrain, Midbrain, and Lumbosacral Spinal Cord of Male Rats☆

This study was designed to determine the extent to which Fos immunoreactivity (induced either by mating or noncontact sociosexual interaction) and androgen receptor (AR) immunoreactivity are colocalized in brain and spinal cord of male rats. Some males (Mated) were allowed to mate to ejaculation; others (Social Controls) were placed with females but physical contact was prevented by a wire mesh screen; remaining males (Isolated) were placed alone in the test jar for the duration of the test period. After testing, brains and spinal cords were examined for AR and Fos immunoreactivity (ir). PG21 anti-AR and anti-c-fos primary antibodies were visualized by fluorescence microscopy using cyanine-conjugated and fluorescein-conjugated secondary antibodies. In both brain and spinal cord, the number of Fos-ir neurons varied according to group: Mated males > Social Controls > Isolated males. Fos was highly localized in subsets of AR-ir neurons within the medial preoptic nucleus, bed nucleus of the stria terminalis, dorsomedial nucleus of the amygdala, and central tegmental field. Fos was also localized in subsets of AR-ir neurons within the L5, L6, and S1 segments of the spinal cord. Spinal cord concentrations of AR-ir and Fos-ir neurons were greatest in Lamina X, and the vast majority of Fos-ir neurons in the dorsal part of Lamina X were also AR-ir. Thus, in both brain and spinal cord, androgen-sensitive neurons are active during mating, and transmission of sexually relevant information from cord to brain is probably accomplished via hormone-sensitive spinal neurons.

Androgen receptor and mating-induced Fos immunoreactivity are co-localized in limbic and midbrain neurons that project to the male rat medial preoptic area

Two studies were designed to document neuronal colocalization of androgen receptor immunoreactivity and mating-induced Fos immunoreactivity (AR-ir, Fos-ir) in brain of male rats and to examine the extent to which limbic and midbrain neurons that project to the preoptic area are androgen sensitive and activated by mating. Brains from male rats, killed 1 h after ejaculating with receptive females, were examined for Fos-ir and AR-ir and compared with those from control rats not given access to females. PG21 anti-AR and anti-c-fos primary antibodies were visualized by fluorescence microscopy using cyanine-conjugated and fluorescein-conjugated secondary antibodies. In mated males (Expt. 1), Fos-ir and AR-ir were colocalized in neurons of the medial preoptic nucleus (MPN), the dorsal medial amygdala (dMEA), the central tegmental field (CTF), the bed nucleus of the stria terminalis, the anterior hypothalamus, the lateral hypothalamus, and the ventral premamillary nucleus. In Expt. 2, male rats received a unilateral injection of the retrograde tracer FluoroGold (FG) in the preoptic area and four days later were killed after ejaculating with receptive females. Brains were subsequently examined for FG transport, Fos-ir and AR-ir. Fluorogold-containing neurons were present in dMEA and CTF as well as in other hypothalamic and limbic regions known to project to the MPN. In dMEA and CTF, nuclear colocalization of AR-ir and mating-induced Fos-ir was present in a proportion of FG-containing neurons. Sexually relevant information may be carried through the brain by an interconnected network of hormone-sensitive neurons.

Estrogen in the medial preoptic area of male rats facilitates copulatory behavior.

Mating was studied in sexually experienced, gonadally intact male rats assigned to two surgical groups matched on the basis of mean mounting frequency during behavioral screening trials conducted prior to the study. Estradiol (E(2)) was delivered bilaterally into the medial preoptic area (MPO) of experimental males by means of hormone-coated implants, and fadrozole was given sc (0.25 mg/kg/day) via osmotic minipumps to block E(2) formation from testicular testosterone throughout the brain. Control males received blank bilateral implants in the MPO and sc fadrozole. After the completion of behavioral testing, immunocytochemical comparisons of the brains from experimental and control rats were made using the H222 antiestrogen receptor (ER) antibody, whose labeling is inhibited by the presence of E(2). The histology demonstrated that E(2) was confined exclusively to the MPO of experimental males but was absent throughout the brains of controls. In controls, mounting decreased significantly by the 7th day after surgery compared with presurgical levels and did not recover. In contrast, on all postsurgical days, the mounting frequency of the experimental group was significantly higher than that of controls. Although experimental males also showed an initial, significant postsurgical decline in mounting frequency, it recovered completely by the 28th postoperative day. Ejaculations declined significantly after surgery in both groups but, unlike in controls whose performance remained low, ejaculations in experimental males partially recovered and were significantly higher than in controls during the postoperative period. Results showed that ER-containing neurons in the MPO influence male rat copulatory behavior.

Estradiol in the male rat amygdala facilitates mounting but not ejaculation

Mating activates estrogen sensitive neurons in several regions of male rat brain, including the medial amygdala (MEA). Infusion of the aromatase inhibitor, Fadrozole, into the MEA reduced mating, presumably by inhibiting conversion of testosterone (T) to estradiol (E(2)). We investigated whether administering E(2) locally into the amygdala (AMG) would maintain sexual behavior in male rats given systemic Fadrozole to eliminate E(2) elsewhere in the brain. Gonadally intact male rats were divided into two matched groups, based on ejaculatory performance in weekly tests with receptive females. All males received 0.29 mg/kg/day sc Fadrozole and bilateral implants to AMG. E(2)-in-AMG males (N=6 experimentals) received implants tipped with a cured mixture of E(2) in Silastic Medical Adhesive, whereas Vehicle-in-AMG males (N=6 controls) received implants tipped with cured adhesive alone (without E(2)). In E(2)-in-AMG males, postoperative mount and intromission frequency did not differ significantly from pretreatment baseline levels, but ejaculation frequency declined significantly (P<.01). Conversely, in Vehicle-in-AMG males, postoperative mounts and intromissions (P<.01) and ejaculations (P<.01) declined significantly. Postoperative mount and intromission frequency of Vehicle-in-AMG males was significantly lower than that of E(2)-in-AMG males (P<.01), but ejaculation frequency did not differ significantly between groups. This suggests that E(2)-sensitive AMG neurons are important for sexual arousal but not ejaculatory performance.

Effects of the Nonsteroidal Aromatase Inhibitor, Fadrozole, on the Sexual Behavior of Male Cynomolgus Monkeys (Macaca fascicularis)

In many vertebrates, castration and hormone replacement and, more recently, the use of aromatase inhibitors, have shown that male sexual activity is mediated by the aromatization of testosterone (T) to estradiol (E2). In macaques, however, the systemic administration of E2, either alone or in combination with androgen, failed either to maintain or to restore the sexual activity of castrated males. The present study examines the effects of administering the nonsteroidal aromatase inhibitor, Fadrozole, either alone or combined with E2, to castrated, T-treated male cynomolgus monkeys at a dose of 0.25 mg/kg/day. This dose inhibited by over 98% the conversion of T to E2 and the subsequent accumulation of the latter in hypothalamic cell nuclei. Castrated males bearing sc Silastic impants of T were each tested with an ovariectomized, E2-treated female partner before, during, and after being given minipumps delivering either Fadrozole or water (240 1-hr tests). Within 2 weeks, Fadrozole significantly reduced ejaculatory activity and male sexual motivation in the absence of changes in plasma T levels, which remained in the upper range for intact males. Additional estradiol treatment produced small but significant increases in ejaculations by three of the six males only, and measures of male sexual motivation remained unchanged (120 tests). The present results, which stand in contrast to our previous findings in macaques, support the view that aromatization of T is important for ejaculatory activity and sexual motivation in a male primate. They also suggest that exogenous E2, which reaches the brain from the systemic circulation, does not fully duplicate the behavioral effects of E2 produced locally in the brain by the aromatization of T.

Annual rhythms in human violence and sexual aggression in the United States and the role of temperature

This study was designed to assess the significance of seasonal changes in assaults, rapes, robberies, and murders committed during 2 to 3 consecutive years in 16 different locations in the US and to relate them both to each other and to seasonal and geographic differences in temperature and photoperiod. The cosinor method of analysis was used to detect annual rhythms in the 4 types of violent crime. Data generated by the cosine functions showed that the annual rhythms in assaults were statistically significant in 12 of 16 locations with maxima between July and September, while annual rhythms in rapes were statistically significant in 14 of 16 locations with maxima also mainly between July and September. Annual changes in robberies were significant in only 5 locations, with maxima between Noverber and December. The annual rhythm for murder was significant in only 1 location. The close temporal relationships between the maxima in assaults and rapes and in the magnitude of the annual fluctuations, together with the fact that rapes are invariably less numerous than assaults, suggest that rapes comprise a subpopulation of assaults. A close statistical association was established between annual assaults and rape rhythms and annual temperature fluctations in the different locations, suggesting that some aspects of human violence may be influenced by exteroceptive factors. There were no significant associations between the mean latitudes of the different locations and the amplitudes of any of the 4 types of violence; the results do not eliminate a role for the photoperiod but fail to demonstrate it.

Evidence for Chemical Communication in Primates

Publisher Summary This chapter discusses the chemical signaling systems in primates and other mammals. The importance of olfaction in the life of numerous primate species under natural conditions is reviewed. The emission and reception of olfactory signals is clearly apparent from the observation of their behavior patterns. The research on the rhesus monkey under laboratory conditions suggested that the male receives information about the hormonal status of his female partner by olfactory pathways. Males made temporarily anosmic failed to detect increases in the attractive properties of their partners, but did so when their olfactory acuity returned. To demonstrate the role of olfactory mechanisms in the behavioral interactions of the pair, it is necessary to conduct experiments to exclude the role of visual signals, and also to exclude the role of afferent stimulation. The olfactory signal appeared to be present in the vaginal secretions of estrogen-treated females. The behaviorally active components are extractable into ether and have acidic properties: they are identified by gas chromatography and mass spectrometry as a series of volatile, short-chain aliphatic acids.

Colocalization of androgen receptors and mating-induced FOS immunoreactivity in neurons that project to the central tegmental field in male rats.

Bilateral lesions of the central tegmental field (CTF) in male rats virtually eliminate mating behavior. This study examined if mating‐induced Fos expression (a measure of neuronal activation) and androgen receptors (AR) are colocalized in brain and spinal cord neurons which project to the CTF. Animals received unilateral injections of the retrograde tracer Fluorogold (FG) in the lateral part of the CTF (CTFl), and 10 days later were killed after ejaculating with females. Brains and spinal cords were examined for FG transport, AR‐immunoreactivity (AR‐ir), and Fos‐immunoreactivity (Fos‐ir). AR‐ir and Fos‐ir were visualized with fluorescence microscopy using cyanine‐conjugated and fluorescein‐conjugated secondary antibodies. The CTFl received projections from AR‐containing neurons in forebrain structures (bed nucleus of stria terminalis, medial preoptic area, lateral and ventromedial hypothalamus), in the central amygdala and various mid‐ and hindbrain structures (dorsolateral tegmentum, superior and inferior colliculi, pedunculopontine nucleus), and in the lumbosacral spinal cord (lamina X). Some of the AR‐containing neurons in bed nucleus of stria terminalis and in the dorsal part of the medial preoptic area with projections to the CTFl were activated by mating. Most AR‐containing neurons in spinal lamina X with projections to the CTFl were also activated by mating. Information from spinal cord and pontine nuclei and from outputs descending from the forebrain may be relayed in the CTFl. Thus, as part of a network of hormone‐sensitive neurons linking brain and spinal cord mechanisms for mating, the CTFl could participate in the integration of visceral and somatic information relevant for sexual behavior. J. Comp. Neurol. 408:220–236, 1999.

Behavioral responses to Depo-Provera, fadrozole, and estradiol in castrated, testosterone-treated cynomolgus monkeys (Macaca fascicularis) : The involvement of progestin receptors

Sexual motivation and behavior decreased in male cynomolgus monkeys given either Depo-Provera (medroxyprogesterone acetate, MPA), which reduces androgen uptake by brain, or the nonsteroidal aromatase inhibitor, Fadrozole, which virtually eliminates the conversion of testosterone (T) to estradiol (E2) in brain. This suggested that both unchanged T and E2 are important for the control of male primate sexual behavior, but combined treatment with MPA and Fadrozole did not have the anticipated summatory effects in intact males: the behavioral decrements when MPA-treated males were given Fadrozole were about half those observed when Fadrozole was given alone. The present study tested the hypothesis that Fadrozole suppressed the behavioral effects of MPA by preventing the induction by E2 of progestin receptors in the brain to which MPA binds. Eight castrated, T-treated males were each tested with an estrogenized female i) during baseline, ii) during MPA treatment, iii) during treatment with MPA and Fadrozole together, and iv) with E2 treatment added to condition (iii) (256 1-h behavior tests). All dosages were those used in previous studies. Sexual motivation, as reflected in mounting attempts and mounting attempt latencies, was further diminished by E2 treatment in males receiving both MPA and Fadrozole, but ejaculatory activity was not changed. Immunocytochemistry demonstrated that the distributions of progestin and androgen receptors were little affected by MPA treatment, and that progestin receptor immunoreactivity was almost completely abolished in the brains of males receiving both MPA and Fadrozole but present in those receiving additional E2 treatment, findings that supported the hypothesis.