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Although aromatase is expressed in both male and female brains, its functional significance in females remains poorly understood. In female quail, sexual receptivity is activated by estrogens. However it is not known whether sexual motivation is similarly estrogen-dependent and whether estrogens locally produced in the brain contribute to these behavioral responses. Four main experiments were designed to address these questions. In Experiment 1 chronic treatment of females with the anti-estrogen tamoxifen decreased their receptivity, confirming that this response is under the control of estrogens. In Experiment 2 chronic treatment with tamoxifen significantly decreased sexual motivation as treated females no longer approached a sexual partner. In Experiment 3 (a) ovariectomy (OVX) induced a significant decrease of time spent near the male and a significantly decreased receptivity compared to gonadally intact females, (b) treatment with testosterone (OVX+T) partially restored these responses and (c) this effect of T was prevented when estradiol synthesis was inhibited by the potent aromatase inhibitor Vorozole (OVX+T+VOR). Serum estradiol concentration was significantly higher in OVX+T than in OVX or OVX+T+VOR females. Together these data demonstrate that treatment of OVX females with T increases sexual motivation and that these effects are mediated at least in part by non-gonadal aromatization of the androgen. Finally, assays of aromatase activity on brain and peripheral tissues (Experiment 4) strongly suggest that brain aromatization contributes to behavioral effects observed here following T treatment but alternative sources of estrogens (e.g. liver) should also be considered., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Steroids modulate the transcription of a multitude of genes and ultimately influence numerous aspects of reproductive behaviors. Our research investigates how one single steroid, testosterone, is able to trigger this vast number of physiological and behavioral responses. Testosterone potency can be changed locally via aromatization into 17β-estradiol which then activates estrogen receptors of the alpha and beta sub-types. We demonstrated that the independent activation of either receptor activates different aspects of male sexual behavior in Japanese quail. In addition, several studies suggest that the specificity of testosterone action on target genes transcription is related to the recruitment of specific steroid receptor coactivators. We demonstrated that the specific down-regulation of the coactivators SRC-1 or SRC-2 in the medial preoptic nucleus by antisense techniques significantly inhibits steroid-dependent male-typical copulatory behavior and the underlying neuroplasticity. In conclusion, our results demonstrate that the interaction between several steroid metabolizing enzymes, steroid receptors and their coactivators plays a key role in the control of steroid-dependent male sexual behavior and the associated neuroplasticity in quail., (Copyright © 2013 Elsevier Inc. All rights reserved.)

During the breeding season, testosterone in male birds is often linked to some secondary sexual ornaments, courtship behaviors, and intrasexual aggression. I examined the effect of castration on plumage expression in Gambel's Quail (Callipepla gambelii), a species in which males are highly ornate, and in Scaled Quail (C. squamata), an unornamented species. Using male pairs, each consisting of a castrate and a control, I also assessed whether castration affected (1) the behavior of males, (2) the mating decisions of females, or (3) the outcome of male-male competition. Castration did not alter the plumage of male Gambel's or Scaled Quail. In these species, and some other members of the avian order Galliformes, production of ornate plumage appears to be independent of testosterone. In contrast, castration reduced or eliminated courtship behaviors. Females almost never preferred castrated individuals. During male-male competition, castrates also exhibited lower rates of threat behaviors, which appear to be identical to those used during courtship. Castration did not, however, influence the outcome of male-male competition. Castrates of both species exhibited overt aggression (pecks, chases, displacement) and frequently won male contests. Such results suggest that certain types of aggressive behavior may be testosterone-independent. In both Gambel's and Scaled Quail, male body size correlated positively with dominant individuals., (Copyright 2001 Academic Press.)

Although testosterone (T) stimulates aggressive and reproductive behaviors in males of many vertebrate species, it is now known that the full expression of T action in the brain requires aromatization to estradiol (E2) and subsequent interaction of locally formed E2 with nuclear estrogen receptors. In experiments reported here, we used a behavioral test which quantifies the response of an individual male Japanese quail (Coturnix coturnix japonica) to the visual stimulus of a conspecific. We have called this behavior aggression because it shares many features in common with traditional measures of aggression, e.g., predicting dominance and subordinance. Nevertheless, the behavior probably also combines a complex steroid-sensitive masculine behavior. The advantage of this test is that it allows the discrimination of individual differences in masculine behavior but avoids fighting and sexual encounters per se, thereby reducing effects of learning, a problem with previous tests of avian aggression. In addition, this test has been applied usefully to identify neuroendocrine correlates to male behavior. Using this test, the arousal of reproductively inactive males (hereafter referred to as aggression) is activated by administration of T or estradiol benzoate (EB), but not by 5 alpha-dihydrotestosterone (DHT). T-induced aggression was blocked by the aromatase inhibitor 4-hydroxyandrostenedione (OHA), an effect partially reversed by treatment with EB. In addition, OHA or the estrogen receptor blocker CI-628 reduced aggressiveness of reproductively active males whereas the androgen receptor blocker flutamide had no effect. Results with the 5 alpha-reductase inhibitor N,N-diethyl-4-methyl-3-oxo-4-aza-5 alpha-androstane-17 alpha-carboxyamide (4-MA) were equivocal. Additionally, treatment of reproductively inactive quail with T or E2 but not DHT increased aromatase activity in the hypothalamus-preoptic area (HPOA). We conclude, therefore, that T to E2 conversion is essential for the activation of aggressiveness in this species. Although locally formed estrogen exerts its effects on aggression in part by increasing activity of aromatase per se, analysis of the time course of behavioral induction or suppression by the various treatments suggests that the response has multiple components, including both short latency, receptor-independent and long latency, receptor-dependent events.

A series of 4 experiments was designed to study the relationships between the activity of the aromatase (AA) in the preoptic area (POA) and the activation by testosterone (T) of copulatory behavior in gonadectomized male and female Japanese quail. The induction of AA by T in the POA is dose- and time-dependent. Levels of AA seen in sexually mature males are restored in castrated birds by a treatment with 20 to 40 mm silastic T capsules which produce physiological levels of steroid in the plasma. The minimal dose of T (10 mm implant) which reliably restores copulatory behavior approximately doubles the AA in the POA. The induction of AA is significantly larger in males than in females. A significant increase in AA is observed within 16 hours after the start of the treatment with T and the induction is maximal after 48 hours. Activation of copulatory behavior follows a similar time course but occurs with a delay of 24-48 hours. These results thus suggest that, in male quail, the activity of the aromatase in the POA is a limiting factor in the activation of copulatory behavior. This idea is confirmed by direct experimentation using an aromatase inhibitor, androstatrienedione (ATD). If T-treated birds receive at the same time silastic implants filled with ATD, the activation of behavior is suppressed for at least one week. This behavioral inhibition is, as expected, accompanied and very probably caused by the inhibition of the aromatase activity in the preoptic area and anterior hypothalamus. No increase of enzyme activity over the level seen in castrates was actually detected during the first 8 days of exposure to T. A moderate increase in AA was seen on day 16 and is probably responsible for the behavioral activation which was observed at the end of the experiment.

In quail, the hypothalamus enzymatically transforms testosterone (T) into estradiol (E2), 5 alpha-dihydrotestosterone (5 alpha-DHT), and 5 beta-dihydrotestosterone (5 beta-DHT). During the embryonic life, the 5 beta-reductase activity is very high, which probably protects the brain of males from being behaviorally demasculinized by their endogenous T. 5 beta androstanes are inactive androgens. The decrease of 5 beta reductase with age during sexual maturation corresponds to a potentiation of the effects of T as shown by experiments that compared the effects of T and 5 alpha-DHT in adult and young quail. T metabolism is also involved in the activation of male behavior in the adult. T aromatization is probably essential for behavioral activation, but nonaromatizable androgens such as methyltrienolone, and to some extent 5 alpha-DHT, can also stimulate sexual behavior in castrates. These enzymatic activities show a clear neuroanatomical localization and are sexually dimorphic. Males produce more active metabolites (E2, 5 alpha-DHT) than females, which could explain the male's greater sensitivity to T treatments. It thus appears that T metabolism is involved in the differentiation and activation of behavior in quail.

DNA polymerase alpha and beta activities were studied and compared in the cloacal gland and the uropygial gland (sebaceous-like gland) taken from castrated quails after testosterone administration. In the cloacal gland, testosterone induced an important increase in DNA polymerase alpha while DNA polymerase beta was not found to be altered. In the uropygial gland, testosterone induced a cyclic variation of DNA polymerase alpha activity which may relate to the holocrine function of this gland.

1. Testosterone reduces the haemolytic action of Ekatin on the morphotic blood elements by accelerating maturation of erythroblastic cells. 2. Testosterone, by accelerating the metabolism of the pesticide causes a defective defence processes of the organism by decreasing the number of macrophages. 3. Intoxication of androgenised birds with Ekatin leads to a breakdown of the systemic adaptive mechanisms by rapid and early switching on the adrenal cortex in response to the stress caused by poisoning.

Mixed cell preparations (theca plus granulosa) prepared from the hierarchy of follicles of quails ovaries were incubated under defined conditions with or without the addition of ovine luteinizing hormone (oLH), ovine follicle stimulating hormone (oFSH), theophylline, cycloheximide, or dibutyryl cyclic adenine monophosphate (db cAMP); or in the presence of androstenedione or testosterone as aromatizable substrate. Steroids secreted into the medium during the 4-hr incubation period were assayed by radioimmunoassay. Cells from the largest follicles (F1) secreted predominantly progesterone, were stimulated by LH and db cAMP, and the response was potentiated by theophylline, but FSH had no stimulatory effect. The F1 cells showed increasing basal and LH-stimulated responses between 18 and 12 hr before the next expected oviposition. Cells from the smaller follicles (F3 and F4) secreted predominantly estrogens, and were stimulated by FSH but not by db cAMP and only to a small extent by theophylline. Addition of androstenedione (10(-7) M) or testosterone (10(-7) M) enhanced estrogen secretion, which was further raised by the simultaneous addition of FSH. These results confirm previous reports on the sites of steroid secretion within quail follicles and suggest that while the action of LH on the cells from F1 follicles may be mediated in part through the adenylate cyclase system, the action of FSH on the smaller follicles may be substantially independent of cAMP.

In 2 independent experiments, we measured and correlated in maturing male Japanese quail the individual variations in sexual and aggressive behavior, cloacal gland size, testes weight, plasma testosterone concentrations and intracellular testosterone metabolism by hypothalamus and cloacal gland. Cloacal gland area was only weakly related to plasma testosterone levels but was positively correlated with the production of active androgenic metabolites and negatively related to the production of 5 beta-reduced androgens (inactive) in the glandular tissue. Several measures of behavior were correlated with aspects of the testosterone metabolism in the anterior hypothalamus. In both experiments, the behavior of the birds was also strongly correlated with their testes weight and their cloacal gland area but weakly or not at all with their plasma testosterone levels. These studies suggest that testosterone metabolism is involved in the control of hormone action in maturing animals.

The uropygial gland of the male quail is a sebaceous-like, androgen-dependent structure. The waxes secreted by this gland consist of fatty acids esterified by alkane-2,3-diols. In adult male quail the relative concentrations of all fatty acids were not affected by castration and testosterone treatment; but in contrast, the relative concentration of dodecane diol was found to be correlated with the androgen levels. In castrated quail administration of testosterone induced an increase in the dodecane diol percentage which was blocked by protein-synthesis inhibitors. Furthermore, the effects of testosterone were found to be neutralized by the administration of cyproterone acetate. Consequently it may be stated that the relative concentration of dodecane diol is indeed a very good marker for androgenicity in the uropygial gland of the male quail. This experimental model seems well suited for evaluating the stimulators and inhibitors of the activity of the sebaceous gland.

We recently identified a sexually dimorphic nucleus in the preoptic region of the Japanese quail, the medial preoptic nucleus (POM), which is significantly larger in males than in females. In the present study, we investigated the hormonal control of this morphological neuroanatomical difference and the possible relationships between the sexual dimorphism in POM volume and in copulatory behavior. Treatments which are known to affect sexual behavior were thus applied to different groups of birds and the POM volume was then measured. In one experiment, male and female quails were either gonadectomized, gonadectomized and treated with testosterone or left intact. The larger size of the POM in males was confirmed and treatments significantly affected the nucleus size which was decreased by gonadectomy and restored by testosterone treatment in both sexes to a level similar to that seen in intact males. In two other experiments, eggs were injected with estradiol benzoate on day 9 of incubation and the POM volume was measured in adulthood either in intact birds or in gonadectomized birds receiving a replacement therapy with testosterone. Despite the fact that estradiol benzoate treatment completely suppressed copulatory behavior, it did not affect the volume of the POM or slightly increased it. These data thus show that the POM volume is controlled by testosterone levels in adulthood and could thus be an interesting model for the study of the effects of steroids on the brain.

The purpose of these experiments was to compare the behavioural and morphological effects of exogenous sex hormones in gonadectomized quail (Coturnix coturnix japonica) with those in quail having regressed gonads as a result of exposure to short days. In Expt 1, male quail were assigned to one of three treatment groups: (1) intact, exposed to 16 h light: 8 h darkness and injected with oil (group 16L); (2) gonadectomized, exposed to 16 h light: 8 h darkness and injected with 2-5 mg testosterone propionate (TP)/day (group 16L-castrated); and (3) intact, exposed to 8 h light: 16 h darkness, and injected with 2-5 mg TP/day (group 8L). Groups 16L-castrated and 8L responded similarly to testosterone, copulating with equal frequency and rapidity after the same number of days of treatment, and also developing proctodeal (foam) glands of a similar size. Only on day 7 of testosterone treatment did the results for these two groups differ. By day 14, the behaviour of both groups resembled that of the 16L birds. In Expt 2 female quail were assigned to the same three treatment groups, except that the hormone treatment was 25 mug oestradiol benzoate/day. Group 8L became sexually receptive sooner than the 16L-ovariectomized quail, but by day 13 both groups had oviducts of similar size, were equally receptive, and were as receptive as the 16L females. The results suggest that the effects of photoperiod on sexual behaviour in this species are mediated largley, if not wholly , by the gonads. They also suggest exposure to short days and surgical gonadectomy are rather similar experimental procedures in the quail.

Capons from high (HML) and low (LML) mating lines of Japanese quail and chickens received testosterone propionate, 5 alpha-dihydrotestosterone (5 alpha-DHT) or estradiol benzoate injections after which copulatory behavior was observed during exposure to live females and to a female model. With live females, the testosterone-treated HML capons mated significantly more than the other HML groups. All LML capons mated infrequently and at comparable levels. When tested with the model, estrogen-treated LML capons mated significantly more frequently than testosterone-treated LML capons, suggesting testosterone aromatization rate was affecting mating activity. This possibility was tested with HLM and LML intact cocks and capons receiving silastic implants of either testosterone, 5 alpha-DHT or a combination of estradiol and 5 alpha-DHT (E + DHT). Mating activity of intact HML cocks, T-treated, and E + DHT-treated HML capons were similar, and all groups mated significantly more than the control or 5 alpha-DHT-treated capons. There were no significant differences between any of the LML groups. The data suggested that limited testosterone aromatization was not the cause of relatively reduced sexual activity in the LML males.

Previous research had shown that reproductively mature male Japanese quail maintained on a 16:08 light/dark (16L:8D) photoperiod spend about 75% of their time throughout daylight hours near a window that provides visual access to a female conspecific. In contrast, females do not display a corresponding tendency to remain near male conspecifics. In Experiment 1, I demonstrated that the social proximity behavior of male Japanese quail declines significantly when photostimulation is restricted to 2 hr daily (02:22 light dark; 2L:22D), but can be restored by reinstituting the 16L:8D schedule. Changes in the photoperiod produced corresponding changes in the size of the cloacal gland, and androgen-dependent organ. The low levels of social proximity behavior and cloacal gland size of males maintained on short daily exposures to light (2L:22D) also could be reversed by sc implants of testosterone (Experiment 2), and this recovery was to some extent sensitive to testosterone dose (Experiment 3). The present studies indicate that social proximity behavior in male Japanese quail is androgen dependent and provide a behavioral assay for neurohormonal studies of sexual behavior that does not depend on brief phasic responses.

Plasma concentrations of progesterone, testosterone, estradiol, and estrone were measured by radioimmunoassay in female Japanese quail 22 to 70 days of age. The birds were raised on long (16 hr light per day) or short (8 hr light per day) photoperiods. Only the birds raised on the long photoperiod layed eggs, starting between 42 and 49 days of age. The concentration of each reproductive steroid increased between 28 and 35 days of age, but the increases were more substantial in the birds raised on long days than in the birds raised on short days. Changes in plasma progesterone and testosterone concentrations showed similar trends: the concentrations increased before the onset of lay and were maintained at a high level in the laying birds. In contrast, plasma concentrations of estradiol and estrone increased to a peak at 35-42 days and declined once egg laying became established. The short-day birds also showed increased plasma estrogen levels, maximal at 35-42 days, with estrone being at a higher concentration than estradiol. The increases in plasma steroid levels occurred during the period of rapid growth of the ovaries and oviducts in the long-day birds, although in the short-day birds the growth of these organs was much less substantial.

Three experiments were carried out to study whether differences in the intracellular metabolism of testosterone (T) can explain sexually differential responses to T in Japanese quail. In the first experiment, a series of dose-response curves in which length of Silastic testosterone implants was related to effects on several behavioral and physiological variables was established. In Experiment 2, adult males and females were assigned to six experimental groups: intact males and females (I-males and I-females), castrated males and females implanted subcutaneously with 40-mm Silastic implants of T (T-males and T-females), and castrated males and females without hormone treatment (CX-males and CX-females). No CX-bird (male or female) and no I-female exhibited male sexual behavior. However, I-males and T-males regularly copulated during the behavioral tests. No crowing was ever heard in CX-animals and I-females. T-females crowed less than T-males and their crowing sounded weaker than those of males. The cloacal glands of T-females were less developed than those of males. Radioimmunoassay of T and 5 alpha-DHT showed that T-males and T-females have similar plasma levels of androgens. No striking differences were observed in the way testosterone is metabolized by the pituitary gland and central nervous tissues of males and females. By contrast, the production of 5 alpha-dihydrotestosterone (5 alpha-DHT) and 5 alpha-androstane-3 alpha, 17 beta-diol (5 alpha, 3 alpha-diol) was higher in the cloacal glands of males than in those of females. These sex differences were not detected between T-males and T-females. In experiment 3, the cloacal gland of males produced more 5 alpha-reduced metabolites than those of females. The pituitary gland of females also produced more 5 beta-androstane-3 alpha, 17 beta-diol (5 beta, 3 alpha-diol). In syringeal muscles, the production of 5 beta-dihydrotestosterone (5 beta-DHT) and 5 beta, 3 alpha-diol was higher in females compared to males.

In the Japanese quail gonadal steroids can depress plasma levels of LH and FSH. Since it is now accepted that testosterone metabolites may be metabolized to the tissue-active forms of the hormone, the in-vitro tissue incubation has been combined with steroid replaement therapy in vivo to investigate the physiological roles of various testosterone metabolites as inhibitory feedback agents on gonadtorophin secretion in quail. After the incubation of quail pituitary glands for 3 h with labelled testosterone four metabolites could be identified; androst-4-ene-3,17-dione, 5 beta-dihydrotestosterone (5 beta-DHT), 5 beta-androstane-3 alpha, 17 beta-diol and 5 alpha-DHT. No 5 alpha-androstane-3 alpha, 17 beta-diol was found. Quantitatively, androstenedione was the major metabolite and converison of testosterone to 5 beta-metabolites was significantly greater than to 5 alpha-androgens. Hypothalamic and hyperstriatal tissues converted testosterone than to 5 alpha-androgens. Hypothalamic and hyperstriatal tissues converted testosterone to androstendione, 5 beta-DHT and 5 beta-androstane-3 alpha, 17 beta-diol but not to 5 alpha-DHT or 5 alpha-androstane-3 alpha, 17 beta-diol. Gonadotrophin secretion was studied in castrated quail after chronic s.c. implantation of steroid-containing silicone elastomer capsules or acute injection i.m. of steroid in ethanol: saline. Irrespecive of the route of administration seven androgens, listed in descending order of potency, reduced the increased levels of plasma LH: testosterone: 5 alpha-DHT; androstenedione; 5 alpha-androstane-3, 17-dione; 5 alpha-androstane-3 alpha, 17 beta-diol; 5 alpha-androstan-3 alpha-ol-17-one; 5 alpha-androstan-3 beta-ol-17-one. No changes in levels of plasma LH were observed after the administration of 5 beta-DHT, 5 alpha-androstane-3 beta,17 beta-diol, 5 beta-androstante-3 alpha, 17 beta-diol, 5 beta-androstane-3, 17-dione, androst-5-ene-3 beta, 17 beta-diol or androst-5-en-3 beta-ol-17-one. Testosterone and oestradiol-17 beta were effective in inhibiting secretion of both LH and FSH in young photostimulated quail and completely blocked testicular growth: 5 alpha-DHT inhibited only the release of LH and testicular growth was unaffected.

This experiment was performed on two groups of male Japanese quail. One had been maintained in our laboratory as a closed colony (S-group), and the other had been obtained from a commercial source (R-group). Different responses of gonadal function were found between two groups following either testosterone treatment or exposure to short days. Immature birds of these groups responded to long days with rapid gonadal growth, but after sexual maturity, exposure to short days for 3 weeks induced testicular atrophy only in S-group. Involvement of the feedback effect of androgen in the photoperiodic response was then examined. Under long-day conditions, intraperitoneal placement of testosterone propionate (TP)-filled Silastic tube for 2 weeks decreased testicular weights in S-group but not in R-group. Apparently, sensitivity to short days is closely correlated with sensitivity to testosterone in the adult male. By bilateral enucleation, quail of S-group became less sensitive to both gonad inhibitory effect of short days and the negative feedback effect of TP. These results suggest that the photoperiodic mechanisms that are primarily mediated by the retinal system play a role in altering sensitivity to steroid feedback at the hypothalamus.

The effects of antioestrogens, antiandrogens and of various inhibitors of testosterone metabolism on testosterone metabolism in the quail hypothalamus and cloacal gland were studied by an in-vitro radioenzymatic assay. It was found that antioestrogens and antiandrogens generally had little or no effect on aromatase and 5 alpha- and 5 beta-reductases of testosterone, except when used at very high doses. The 5 alpha-reductase inhibitor, 17 beta-N,N-diethylcarbamoyl-4-methyl-4-aza-5 alpha-androstan-3-one, inhibited both 5 alpha- and 5 beta-dihydrotestosterone production without markedly affecting aromatase activity. Surprisingly, the aromatase inhibitor, 1,4,6-androstatriene-3,17-dione, inhibited not only the production of oestradiol but also that of 5 beta-dihydrotestosterone and, to a lesser extent, 5 alpha-dihydrotestosterone. These unexpected properties should be taken into account when interpreting the results of in-vivo experiments using these compounds.

Photosensitivity of the quail subcommissural organ (SCO) is demonstrated. When the birds are submitted to continuous light the SCO cells appear depleted in secretory material and the enzyme activities investigated are of greater intensity. Such a diminished secretory content is observed in birds exposed to short daylength when they are injected with testosterone derivatives. Besides, injections of antiandrogenic compounds in continuously illuminated quails abolish the decrease of the secretory material induced by photostimulation. These results indicate that the SCO responsiveness to light appears to be mediated by some hormonal mechanism. Moreover it is more than a generalized metabolic effect: the response to antiandrogen injections suggests indeed that the SCO is a likely target organ for hormonal action.

Testosterone causes a significant body temperature decrease in male quails. Oxygen consumption/g b. wt remains the same, however.