Androgen and estrogen stimulation of ornithine decarboxylase activity in mouse kidney

In the present work, the activity of mouse renal ornithine decarboxylase (ODC) from CBA female mice was used as a biological marker to detect (anti)androgenic activity of different groups of endocrine disruptors and steroids. Daily injections of testosterone or dihydrotestosterone (DHT) into 60 day old female mice for 4 days increased renal ODC activity in a dose-dependent manner that reached up to 100-fold (testosterone) or 250-fold (DHT) above the baseline when the highest dose, 200 microg/mouse, was used. Administration of flutamide concurrently with testosterone (75 microg/mouse) caused a potent decrease of ODC induction in a dose-dependent manner, suppressing the enzyme activity at the doses of 0.1 and 0.5 mg/mouse by about 88 and 95%, respectively. In contrast, estradiol at the doses of 0.5 and 1 mg/mouse induced a significant stimulation of renal ODC activity in a dose-dependent manner when it was given alone or in combination with testosterone. Using a sensitive increase in ODC activity in response to androgens as an end point, we did not detect an antiandrogenic effect of several antiandrogens, such as cyproterone acetate, spironolactone, p,p'DDE and vinclozolin. Also, none of these antiandrogens were able to change the basal level of renal ODC activity, with the exception of cyproterone acetate that at a dose of 0.1 mg/mouse stimulated ODC activity. The data obtained suggest that mouse renal ODC from CBA females is not strictly androgen-specific and cannot be used for estimation of antiandrogenic effects of compounds having an affinity to different types of receptors.     

Effects of the antiandrogens cyproterone acetate and flutamide on male reproductive behavior in a lizard (Anolis sagrei)

This study examined the effects of the antiandrogens cyproterone acetate (CA) and flutamide (F) on male reproductive behavior in the lizard Anolis sagrei. Reproductively active males were implanted with subcutaneous pellets of either CA, F, or placebo (P). Pellets delivered CA and F at a constant rate of 0.1 mg/day. Three weeks after implantation, males were tested with stimulus males and two days later with stimulus females. Cyproterone acetate inhibited aspects of male aggressive and sexual behavior, and reduced testis weight and size of the renal sex segment. Plasma testosterone (T) levels in CA-treated males were not significantly different than those of P-treated males. Flutamide did not inhibit aggressive or sexual behavior, but did decrease testis weight as well as the size of the renal sex segment. Plasma T levels were significantly higher in F-treated males than in P-treated males. These data suggest that CA, an antiandrogen with antigonadotropic activity, may be used to inhibit reproductive behavior in male lizards.     

Stimulatory effects of 5 beta-dihydrotestosterone on the sexual behavior in the domestic chick

During in vitro incubations, brain tissues of chicks transform testosterone mainly into 5β-reduced androgens including 5β-dihydrotestosterone (β-DHT). Although β-DHT is generally believed to have no androgenic effects, we showed recently that it stimulates sexual behavior in young chicks during hand thrust tests. This result was confirmed during three experiments presented here and the possible interactions of β-DHT with the endogenous sex steroids have been analyzed. There is no synergism between β-DHT and estradiol in the induction of sexual behavior in the chicks. β-DHT is still partly active in chicks injected with the antiandrogen, cyproterone acetate in doses sufficient to block all action of the endogenous testosterone. β-DHT is also active in castrated chicks. The effects of β-DHT on the sexual behavior thus do not seem to depend on an interaction with other sex steroids and the reasons why it is active in chicks during hand thrust tests and not in other test situations in other birds are briefly discussed.     

Effects of steroidal and non-steroidal antiandrogens on the androgen binding properties of the rat ventral prostate androgen receptor

Steroidal (cyproterone acetate) and non-steroidal (RU23908 and hydroxyflutamide) antiandrogens are able to block testosterone-induced increases in nuclear androgen receptor (AR) in the prostate of 1-day orchidectomized rats, but when given alone, RU23908 and hydroxyflutamide increase nuclear AR (RU23908 greater than hydroxyflutamide) in the same animal model. The increases in nuclear AR induced by antiandrogen alone or with testosterone alone are blocked by cycloheximide 1 h after administration, suggesting that androgen or antiandrogens induce de novo AR synthesis. Concomitant to nuclear AR accumulation, testosterone is able to induce depletion of cytosol and microsomal AR. Blockade of testosterone-induced depletion of microsomal AR, but not of cytosol AR, occurs in the presence of antiandrogens. Cyproterone acetate has a higher relative binding affinity (RBA) for microsomal AR and cytosol AR than RU23908 or hydroxyflutamide. This phenomenon is in good agreement with the degree of inhibition by these compounds of the association rate of androgen for the microsomal AR. This correlation between RBA and inhibition of the initial rate of hormone binding to the receptor is not found for cytosol AR. The results show that antiandrogens are not 'pure' antagonists of androgen action and they are potent agonists in the absence of testosterone. Furthermore, testosterone alone or antiandrogens per se regulate AR levels acutely by protein-synthesis dependent mechanisms of action, in rat ventral prostate.     

Territorial aggression and song of male European robins (Erithacus rubecula) in autumn and spring: effects of antiandrogen treatment

Male robins aggressively defending a feeding territory in winter have low levels of testosterone, while males defending a breeding territory in spring have elevated levels of testosterone. Song is an integrated part of territorial defense during both phases. We investigated whether testosterone is involved in the expression of these behaviors by treating free-living and captive male robins during both phases with the antiandrogen flutamide. Results suggest that, similar to species in which territoriality is restricted to the reproductive phase, aggressive defense of a breeding territory by male robins is facilitated by androgens. Territorial defense during the nonbreeding season, however, does not require androgenic activity. Singing frequency, on the other hand, was not significantly reduced during either phase by flutamide application. Since the quality of male song changes with season we suggest that the incorporation of sexual signals into male song during breeding depends on testosterone, but that the year-round production of territorial "keep out" signals is independent of testosterone.     

Effect of antiandrogen cyproterone acetate on the action of testosterone on mouse kidney.

The loss of endogenous testosterone in castrated male mice leads to a marked decrease in seminal vesicle and kidney tissue weight. 21 days' administration of exogenous testosterone abolished the effect of castration on the seminal vesicles and kidney tissue. The antiandrogen cyproterone acetate produced significant changes in the target tissue for androgens, i.e. in the seminal vesicles. In every case it blocked the action of both exogenous and endogenous testosterone on the seminal vesicles, but failed to block the "renotropic" action of testosterone, expressed as relative kidney weight. Contrary to its effect on the seminal vesicles, it did not influence relative kidney weight in normal animals. It likewise did not block the effect of exogenous testosterone on kidney tissue. The mechanism of the action of cyproterone acetate in androgen-dependent tissues is known to consist in inhibition of androgen binding to specific cell receptors in the target tissues. Some of the specific androgen receptors in mouse kidney are evidently different in character from those in the accessary sex glands, that being the reason why cyproterone acetate has an antiandrogenic, but not an antirenotropic effect. In agreement with experiments on rats, adrenal weight also decreases in mice after the administration of cyproterone acetate.     

Androgen receptor binding and antiandrogenic activity of some 4,5-secoandrostanes and ring B cyclopropanoandrostanes

Eight androstane derivatives with modified ring A or B (4,5-secoandrostanes and ring B cyclopropanoandrostanes) were assayed in vivo on mice for their antiandrogenic activity and the effect was compared with that of cyproterone acetate. The inhibition of dihydrotestosterone binding to rat prostate cytosol and to human plasmatic sex hormone binding protein was correlated with the in vivo effect. The antiandrogenic activity of 6 alpha,7 alpha-cyclopropano-5 alpha-androstane-3 beta,17 beta-diol was nearly as high as that of cyproterone acetate. The opening of ring A of androgens, such as testosterone or 17 alpha-methyltestosterone, moderately reduced the binding and changed the biological activity to weakly antiandrogenic.     

Androgen-dependent fighting behaviour in male mice

The influence of testosterone propionate, 17 alpha-methyl-testosterone and cyproterone acetate on isolation induced fighting behaviour of mice was studied in a simple testing procedure. Decreased aggressiveness has been established in mature, sexual experienced and isolated male mice both following castration and administration of the antiandrogen cyproterone acetate, respectively. Replacement therapy with testosterone propionate s.c. and 17 alpha-methyl-testosterone p.o. has been shown to restore the decreased level of aggresiveness after castration.     

睾酮对鸟类繁殖影响研究进展

类固醇激素睾酮是影响鸟类繁殖最重要的性激素之一,与鸟类的繁殖行为的各个方面息息相关。睾酮通过影响鸟类的羽色、鸣声等来影响鸟类的配偶选择,同时睾酮可以调节配偶选择和繁殖投入之间的平衡。睾酮水平影响出雏数、出飞数、孵化率成功率等繁殖成效。睾酮还对个体的免疫活性和个体的存活率等产生影响。目前关于睾酮对鸟类繁殖影响的研究大多是通过外源性植入睾酮的方式来改变个体睾酮的浓度,其研究结果也常出现相互矛盾之处,对于自然状态下影响睾酮水平变化的因素尚缺乏了解,睾酮对雌雄鸟在繁殖过程中的影响也不尽相同,有必要继续深入研究。     

Inhibition of androgen binding in human foreskin fibroblasts by antiandrogens

Antiandrogen effects on androgen receptor binding and androgen metabolism were studied in cultured human newborn foreskin fibroblasts. Three different antiandrogens were tested in this system: (a) cyproterone acetate (CA); (b) RU23908; and (c) R2956. CA and R2956 were equipotent inhibitors of androgen binding to its intracellular receptor. The magnitude of this action was nearly twice as great against the endogenous androgen ligands, dihydrotestosterone (DHT) or testosterone (T), than with the synthetic ligand, methyltrienolone (R1881). Whereas the relative binding affinities of CA and R2956 were approximately 5-10 times less than T or DHT, RU23908 was another order of magnitude less effective as an inhibitor of androgen binding. The lower relative binding affinity determined for RU23908 could not be explained on the basis of a requirement for metabolic activation. Subcellular fractionation studies and sucrose density gradient analysis further confirmed the rank order of antiandrogenic potency. None of the antiandrogens influenced the rate or profile of metabolites from cellular metabolism of T or DHT. We propose that cultured human genital skin fibroblasts may serve as a valuable system for the future evaluation of antiandrogens in intact ells under physiologic conditions.     

Behavioural and Physiological Effects of Testosterone Propionate and Cyproterone Acetate in Immature Male Domestic Ducks,Anas platyrhynchos

The behavioural and morphological effects of testosterone propionate and of the anti-androgen cyproterone acetate were studied in male domestic ducks. Testosterone was also measured by radioimmunoassay in the plasma of these birds to relate the behaviour to the actual circulating levels of hormone. Testosterone stimulates sexual behaviour but has few effects on social displays. There is no correlation between the individual variations of plasma testoterone and sexual behaviour.     

Biological manifestations of the andropause

Aging in men is accompanied by signs of decreased virility and fertility: testicular volume, muscle mass, pilosity, sexual activity, daily production of spermatozoa, plasma testosterone levels, the number of Leydig cells and blood supply of the testicles decrease significantly with age. The pulsatility of LH levels in young and elderly men has been studied by taking plasma samples with an interval of 10 minutes for 12 hours. Age alterations at the hypothalamic-pituitary levels of testosterone production and an increase in gonadostat sensitivity to hormonal influences realized via the feedback mechanism are found. Whenever androgen therapy of eugonadal elderly men is considered, a sufficient dosage should be administered, as otherwise, due to the feedback effects, administered androgens, suppressing endogenous secretion might not increase plasma-testosterone levels. Changes in neurotransmitters and neuromodulators play a determinant role in the sexual behaviour of elderly men.     

Towards the mapping of the progesterone and androgen receptors

At a time when the secondary structures of receptor proteins are being predicted from sequence data by modeling techniques, knowledge of the ligand characteristics compatible with high-affinity binding to the receptor and with efficient receptor function is indispensable. We have already compared progesterone receptor (PR) ligands in attempts to map the PR hormone-binding site. In the present study, the relative binding affinities (RBAs) of 33 steroid ligands for the cytosol androgen receptor (AR) of rat prostate, measured in a routine screening system, have been compared. Special emphasis has been given to the effects of modifications (unsaturation, methylation, substitution by halogens) that might influence AR recognition by the ring A carbonyl and also to the consequences of these changes on binding specificity. Nonsteroid antiandrogens are reputed to compete with labelled testosterone (or methyltrienolone) binding to AR. Their RBAs, however, are very low compared to those of steroid antiandrogens. It is feasible that such molecules might occupy and interact with the AR site that binds hormone. The solvent accessible surface of one Anandron conformer is highly similar to that of testosterone and this conformer can be adequately superimposed upon the structure of testosterone and of antiandrogenic Des-A steroid derivatives. The nitro group might assume the role of the ring A carbonyl of steroids; reduction of this group to an amine or a hydroxylamine completely suppresses binding. These observations, however, do not eliminate the hypothesis of interference with AR function, and consequent antiandrogenic activity, by interaction with other (adjacent) sites on AR.     

Castration and antisteroid treatment impair vocal learning in male zebra finches.

Both song behavior and its neural substrate are hormone sensitive: castrated adult male zebra finches need replacement of gonadal steroids in order to restore normal levels of song production, and sex steroids are necessary to establish male-typical neural song-control circuits during early development. This pattern of results suggests that hormones may be required for normal development of learned song behavior, but evidence that steroids are necessary for normal neural and behavioral development during song learning has been lacking. We addressed this question by attempting to eliminate the effects of gonadal steroids in juvenile male zebra finches between the time of initial song production and adulthood. Males were castrated at 20 days of age and received systemic implants of either an antiandrogen (flutamide), an antiestrogen (tamoxifen), or both drugs. The songs of both flutamide- and tamoxifen-treated birds were extremely disrupted relative to normal controls in terms of the stereotypy and acoustic quality of individual note production, as well as stereotypy of the temporal structure of the song phrase. We did not discern any differences in the pattern of behavioral disruption between birds that were treated with either flutamide, tamoxifen, or a combination of both drugs. Flutamide treatment resulted in a reduced size of two forebrain nuclei that are known to play some role unique to early phases of song learning [lateral magnocellular nucleus of the anterior neostriatum (IMAN) and area X (X)], but did not affect the size of two song-control nuclei that are necessary for normal song production in adult birds [caudal nucleus of the ventral hyperstriatum (HVc) and robust nucleus of the archistriatum (RA)]. In contrast, treatment with tamoxifen did not result in any changes in the size of song-control nuclei relative to normal controls, and it blocked the effects of flutamide on the neural song-control system in birds that were treated with both drugs. Castration and antisteroid treatment exerted no deleterious effects on the quality of song behavior in adult birds, indicating that gonadal hormones are necessary for the development of normal song behavior during a sensitive period.     

Quantitative Untersuchungen zu Wechselbeziehungen zwischen Balzaktivität und Testosterontitern bei männlichen Zebrafinken

The determination of hormone titers in small plasma volumes by radioimmunoassay makes it possible to demonstrate the mutual relationship between testosterone titers and sexual behaviour. Testosterone titers were measured after courtship tests in male Zebra Finches to determine whether these levels are influenced by sexual behaviour. The results show that immediately after a courtship test there is a significant difference in hormone levels between sexually experienced and inexperienced birds. Sexual experience can also be shown to influence baseline testosterone titers in male Zebra Finches: experienced ♂♂ have a high baseline level while inexperienced ♂♂ have a low baseline level. In two other behaviour patterns, undirected song and courtship song, it could be demonstrated, that the amount of these behaviours shown is linearly correlated with the amount of testosterone in the blood plasma. Both song types have different hormonal threshold values. The comparison of testosterone titers between different treatment groups can only be made if the groups are maintained under identical conditions, since housing conditions appear to have a very marked influence on testosterone titers. The mechanism of action of testosterone in sexual behaviour is discussed.     

Yolk testosterone, postnatal growth and song in male canaries

Avian eggs contain substantial amounts of maternal yolk androgens, which have been shown to modulate offspring phenotype. The first studies on the functional consequences of maternal yolk androgens have focused on early life stages and their role in sibling competition. However, recent longitudinal studies reported long-lasting effects of maternal yolk androgens on offspring phenotype, mostly concerning traits that are sensitive to androgens. This suggests that maternal yolk androgens could play an important role in sexual selection, since the expression of many male sexual characters is testosterone-dependent. Using male canaries as a model, we examined the consequences of an experimental elevation of yolk testosterone concentrations on early development as well as long-lasting effects particularly on song, which is one of the most important sexual characters in male songbirds. Elevated yolk testosterone concentrations inhibited male growth, possibly in interaction with an existent ectoparasite exposure. Males hatched from testosterone-treated eggs (T-males) did not have enhanced competitive skills, in contrast to previous studies. The elevation of yolk testosterone concentrations delayed song development but did not affect adult song phenotype. This is intriguing, as yolk testosterone possibly induced developmental stress, which is known to reduce song quality. We hypothesize that yolk testosterone has either no direct effect on adult song phenotype, or that positive effects are merged by the negative effects of developmental stress. Finally, females mated with T-males invested more in their clutch indicating that females either assess T-males as more attractive (differential allocation hypothesis) or compensated for lower offspring viability (compensation hypothesis).     

Interspecific variation in egg testosterone levels: implications for the evolution of bird song

Although interspecific variation in maternal effects via testosterone levels can be mediated by natural selection, little is known about the evolutionary consequences of egg testosterone for sexual selection. However, two nonexclusive evolutionary hypotheses predict an interspecific relationship between egg testosterone levels and the elaboration of sexual traits. First, maternal investment may be particularly enhanced in sexually selected species, which should generate a positive relationship. Secondly, high prenatal testosterone levels may constrain the development of sexual characters, which should result in a negative relationship. Here we investigated these hypotheses by exploring the relationship between yolk testosterone levels and features of song in a phylogenetic study of 36 passerine species. We found that song duration and syllable repertoire size were significantly negatively related to testosterone levels in the egg, even if potentially confounding factors were held constant. These relationships imply that high testosterone levels during early development of songs may be detrimental, thus supporting the developmental constraints hypothesis. By contrast, we found significant evidence that song-post exposure relative to the height of the vegetation is positively related to egg testosterone levels. These results support the hypothesis that high levels of maternal testosterone have evolved in species with intense sexual selection acting on the location of song-posts. We found nonsignificant effects for intersong interval and song type repertoire size, which may suggest that none of the above hypothesis apply to these traits, or they act simultaneously and have opposing effects.     

Immune challenge mediates vocal communication in a passerine bird: an experiment

Secondary sexual characters may have evolved in part to signalresistance to parasites. Avian song has been hypothesized tobe involved in this process, but the role of parasites in modulatingacoustic communication systems in birds remains largely unknown,owing to lack of experiments. We studied the relationship betweenparasitism, testosterone, song performance, and mating successin male collared flycatchers (Ficedula albicollis) by experimentallychallenging their immune system with a novel antigen. We predictedthat a challenge of the immune system would reduce song performance,and that this reduction would be conditional on the size ofa visual sexual signal, the forehead patch that was previouslyfound to reflect resistance. An antagonistic linkage betweentestosterone and immune function would predict that a challengeof the immune system should suppress testosterone level. Animmunological treatment by sheep red blood cells (SRBCs) triggereda decrease in body mass, testosterone level, and song rate,but other song traits were not significantly affected by theantigen challenge. Initial testosterone level was associatedwith forehead patch size and all song traits except song rate.SRBC injection caused stronger reduction in song rate amongmales with smaller forehead patches, and the change in songrate was also predictable by song features such as strophe complexityand length. We show that song rate and other song characteristicsmay be important cues in male-male competition and female choice.These results suggest that parasite-mediated sexual selectionhas contributed in shaping a complex acoustic communicationsystem in the collared flycatcher, and that testosterone mayplay an important role in this process. Parasitism may drivea multiple signaling mechanism involving acoustic and visualtraits with different signal function.