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J Balthazart

Publications and source records attributed to J Balthazart.

At least 163 records · Page 9Linked to original sources

A dorsomedial subdivision within the nucleus intercollicularis identified in the Japanese quail (Coturnix coturnix japonica) by means of alpha 2-adrenergic receptor autoradiography and estrogen receptor immunohistochemistry.

The nucleus intercollicularis is an important site in the control of vocalization in birds. In oscines, a subregion of the nucleus intercollicularis called the dorso-medial intercollicular nucleus appears to play a key role in this process because it receives the majority of the projections from the nucleus robustus archistriatalis and sends most of the projections to the motor nucleus of the hypoglossal nerve. In this paper, we present neurochemical studies of the nucleus intercollicularis in the Japanese quail which suggest the presence of heterogeneity within this structure. One rostral band contains high densities of cholinergic muscarinic receptors identified by quantitative autoradiography using tritiated N-methylscopolamine as the ligand. A caudal dorso-medial region is specifically labeled by estrogen receptors identified using immunocytochemistry and by alpha 2-adrenergic receptors which were quantified by autoradiography using tritiated para-amino-clonidine. This latter subregion is possibly equivalent to the dorso-medial intercollicular nucleus of oscines. Additional track-tracing studies should be performed to confirm this homology. The coexistence of estrogen and alpha 2-adrenergic receptors within the same structure suggests important functional connections between steroid action and catecholaminergic systems in the brain.

Animals↗

Sexual differentiation in quail: critical period and hormonal specificity.

There is a discrepancy between results showing that male quail are demasculinized by exogenous estrogens only if the treatment is given before Day 12 of egg incubation and results showing that ovariectomy of females after hatching still affects their sexual differentiation which leads to the conclusion that female demasculinization by ovarian estrogens is a continuing process extending into posthatching life. The first experiment was performed to test different models which have been proposed to reconcile these apparently contradictory results. Male and female quail were treated with 0, 5, or 25 micrograms of estradiol benzoate (EB) on either Day 9 or Day 14 of embryonic life. Birds were castrated at the age of 4 days to avoid the confounding effects of postnatal gonadal hormones and were treated as adults with testosterone (T). Whereas EB-treatment demasculizined sexual behavior and cloacal gland growth of males when administered on Day 9, it was without effect on Day 14. This result confirms the presence of a "critical period" for sexual differentiation of behavior in embryonic life. However, the time course of sexual differentiation and the sensitivity to the demasculinizing actions of estrogens were not the same for different behavioral and morphological characteristics. Some dependent variables such as plasma levels of luteinizing hormone and crowing were still affected by the EB treatment on Day 14. These results show that the whole process of demasculinization is not retricted to the "critical period" ending on Day 12 of incubation. A second experiment was performed to determine if 5 beta-dihydrotestosterone (5 beta-DHT), a metabolite of testosterone, also exerts demasculinizing effects during embryonic life. A large dose of 5 beta-DHT (2 mg/egg) had no effects on behavior and morphology in males if administered on Day 9 of egg incubation. This suggests that 5 beta-DHT, which is a steroid devoid of behavioral effects in the adult bird, is also an inactive compound as far as sexual differentiation of the quail is concerned. The high 5 beta-reductase activity which was previously identified in the hypothalamus of the embryonic quail thus probably plays a protective role. By transforming testosterone into inactive nonaromatizable androgens, it prevents male embryos from being demasculinized by their endogenous testosterone acting through aromatization.

Animals↗

Correlation between the sexually dimorphic aromatase of the preoptic area and sexual behavior in quail: effects of neonatal manipulations of the hormonal milieu.

The aromatase of the preoptic area is significantly more active in males than in females. This sex dimorphism in enzyme activity is still found in birds that have been gonadectomized and treated with a same dose of testosterone. This suggests that the sex difference is not the result of a differential activation by the adult hormonal environment but rather is organized neonatally by steroid hormones. As the central aromatization of testosterone is a limiting step in the activation of copulatory behavior by testosterone, the lower aromatase activity in the preoptic area of females might be responsible, at least in part, for their lower sensitivity to the activating effects of testosterone on behavior. Three experiments were carried out to determine whether early manipulations of the hormonal environment, which are known to differentiate sexual behavior, also affect in a permanent way the aromatase activity in the preoptic area. Injection of estradiol benzoate into male embryos on day 9 of incubation decreased the preoptic aromatase activity in parallel to its demasculinizing effect on behavior. Unexpectedly the same treatment tended to increase enzyme activity in females so that the physiological relevance of the observed enzymatic change remains questionable. In two independent experiments, we confirmed that neonatal ovariectomy of female quail interferes with their behavioral differentiation. Females gonadectomized at 4 days post-hatch showed significantly more male-type sexual behavior as adult in response to testosterone than females gonadectomized at the age of 5 weeks. These experiments also confirmed that the preoptic aromatase activity is higher in males than in females but no evidence for an effect of the age of gonadectomy on the enzyme activity could be obtained. The sex difference and experimental modifications observed in the aromatase activity of the preoptic area were not seen in the posterior hypothalamus demonstrating that these effects are specific. The mechanisms controlling the sex difference in aromatase activity are discussed. The difference might be organized by the action of embryonic steroids as suggested by the changes observed in males injected with estradiol benzoate in egg. Alternatively, activational mechanisms cannot be ruled out at present. In one experiment, the activity of the preoptic aromatase was positively correlated with the sexual activity of the birds.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Changes in the activity of testosterone-metabolizing enzymes in the brain of male and female zebra finches during the post-hatching period.

The kinetic properties of the testosterone-metabolizing enzymes were studied in the hypothalamus of adult and young zebra finches of both sexes. Estradiol, 5 alpha-dihydrotestosterone, 5 beta-dihydrotestosterone and 5 beta-androstane-3 alpha,17 beta-diol were identified as metabolites of testosterone in males and females at different ages between 5 days post-hatch and adulthood. During maturation, the maximum velocity (Vmax) of the aromatase and 5 beta-reductase decreased in males and females while the affinity of these enzymes for the substrate increased (decrease in km). These changes were more pronounced for 5 beta-reductase than for aromatase. The affinity change of the 5 beta-reductase occurred progressively during the post-hatching development between 5 and 30 days and is thus probably a true developmental process. In the case of the aromatase, the change in affinity occurred much later (after 30-40 days) and is thus probably related to the sexual maturation. These kinetic changes during development are directly related to the roles played by testosterone and its metabolites, in particular estradiol, in the differentiation and activation of reproductive behavior in the zebra finch. In particular, the dramatic decrease in 5 beta-reductase activity during sexual maturation should correspond to a potentiation of testosterone action in the brain.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Changes in progesterone metabolism in the chicken hypothalamus during induced egg laying stop and molting.

In the present study, we have established and validated a radioenzyme assay which permits us to quantify progesterone metabolism in the chicken brain. Progesterone metabolism was then studied in five brain areas obtained by microdissection from the telencephalon (part of the lobus paraolfactorius immediately rostral to the preoptic area), the preoptic area, and the hypothalamus. Three metabolites of progesterone were produced in large amounts in these brain regions and were quantified in this study: 5 beta-pregnane-3,20-dione (5 beta-DHP) as well as its metabolite 3 alpha-hydroxy-5 beta-pregnane-20-one (5 beta,3 alpha-ol) and 5 alpha-pregnane-3,20-dione (5 alpha-DHP). The unmetabolized progesterone was also recovered and quantified. The 5 beta-reduction of progesterone (production of 5 beta-DHP and 5 beta,3 alpha-ol) was very active but its 5 alpha-reduction (production of 5 alpha-DHP) was almost absent in the lobus paraolfactorius. An opposite pattern of metabolism was found in the preoptic area and the hypothalamus (higher 5 alpha- but lower 5 beta-reductase activity). The changes in progesterone metabolism in these brain areas were then studied in groups of hens submitted to induced egg laying stop and molting. A significant decrease in progesterone 5 alpha-reduction was found in the median hypothalamus of hens during the period of molt. Simultaneously, the experimental procedures induced significant decreases in the production of 5 beta-DHP by the lobus paraolfactorius, anterior, and medial hypothalamus but induced a significant increase in the production of this metabolite in the preoptic area. These changes are likely to be involved in the control of reproductive functions including sexual behavior and secretion of luteinizing hormone-releasing hormone, and a number of possible causal mechanisms are presented. These should now be tested experimentally especially in view of the very limited information which is now available on the biological effects of the metabolites of progesterone.

3-Hydroxysteroid Dehydrogenases↗

Changes in serum concentrations of steroids during embryonic and post-hatching development of male and female Japanese quail (Coturnix coturnix japonica).

Serum concentrations of testosterone, 5 alpha-dihydrotestosterone, oestradiol and progesterone were measured by radioimmunoassay combined with Celite chromatography in male and female Japanese quail (Coturnix coturnix japonica) during the second half of embryonic life (days 9-17 of incubation) and during the first 5 weeks after hatching. The mean level of each of the four steroids was significantly affected by the age of the birds. An overall effect of sex was detected by analysis of variance only on oestradiol concentrations, with females having higher serum concentrations than males during most of the age range studied. Significant peaks of testosterone and progesterone were also detected around hatching time. These results are consistent with the view that oestradiol is the major hormone implicated in the sexual differentiation of reproductive behaviour in the quail. The relationships between the circulating concentrations of oestradiol during ontogeny and the critical period of differentiation as postulated by currently accepted models is also discussed.

Animals↗

Neuroanatomical distribution of testosterone-metabolizing enzymes in the Japanese quail.

We describe a very sensitive and precise assay which allows one to study the metabolism of testosterone (T) in small brain nuclei dissected out according to the method of Palkovits and Brownstein. With this method, the neuroanatomical distributions of aromatase, and 5 alpha- and 5 beta-reductase activities were studied in adult male quail (Coturnix coturnix japonica). The different enzymes show different neuroanatomical distributions. Production of estradiol-17 beta (E2) was highest in the sexually dimorphic nucleus preopticus medialis (POM). We showed previously that the preoptic aromatase activity is higher in male than in female quail. As the POM is a central and very large structure within the preoptic area, the present results suggest a relationship between the neuroanatomical and the biochemical sex differences. By contrast, the production of 5 alpha-DHT was highest in the lateral hypothalamic area (LHY), the bed nucleus of the pallial commissure (BPC) and the lateral septum (SL). The 5 beta-reductase activity was highest in the lateral septum and in the ventral part of the archistriatum (AV). Moreover, there was a rostral to caudal decrease in 5 beta-reductase activity in the hypothalamus.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Central and peripheral metabolism of 5 alpha-dihydrotestosterone in the male Japanese quail: biochemical characterization and relationship with reproductive behavior.

An in vitro radioenzymatic assay and purification procedure by thin-layer chromatography were used to study the metabolism of dihydrotestosterone (DHT) into 3 alpha- and 3 beta-androstanediols by the brain and cloacal gland of Japanese quail. Kinetic studies showed that these 2 metabolites are produced in a linear fashion with respect to time of incubation for up to 15 min but that they continue to accumulate for up to 4 h. The maximum velocity of these reactions is high (nmol/mg protein/15 min), but the affinities of the enzymes for DHT are low (in the microM range). The enzymatic activities are not evenly distributed in the brain: they are high in the tuberal hypothalamus and lobus parolfactorius but low in the preoptic area and anterior hypothalamus. Enzyme activities are not markedly affected by treatment of the birds with either testosterone or DHT. The activity of these enzymes is lower in the preoptic area and tuberal hypothalamus of DHT-treated birds which display female-directed sexual behavior than in the same brain areas of birds which are sexually inactive. We discuss the relationships between this reductive metabolism of DHT and the activational effects of the steroid on sexual behavior.

Androstane-3,17-diol↗

Sexual differentiation and hormonal control of the sexually dimorphic medial preoptic nucleus in the quail.

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.

Animals↗

Effect of the injection solvent on steroid activity.

The effects of testosterone, 5 alpha-dihydrotestosterone, and androstenedione on cloacal gland and sternotracheal muscle growth and on luteinizing hormone plasma levels were studied in Japanese quail. The steroids were given to the animals in daily intramuscular injections and were dissolved either in propylene glycol or in ethanol:saline (1:1). The three steroids had significant effects on the responses which were measured, but the magnitude of this effect was clearly related to the type of vehicle in which the steroids were dissolved. Furthermore there was a significant interaction between the effects of solvents and of steroid; for example, testosterone was much less androgenic than 5 alpha-dihydrotestosterone when dissolved in propylene glycol, but both had similar effects when dissolved in ethanol:saline. The effects of the mode of administration of different steroids on a given physiological response should be taken into consideration.

Androgens↗

Hormonal control of female sexual behavior in the Japanese quail.

Four experiments were carried out to study the hormonal control of female receptivity and proceptivity in Japanese quail. Both aspects of reproductive behavior can be activated in a dose-dependent manner by injections of estradiol benzoate (EB). Progesterone (P) given in addition to suboptimal doses of EB has little additional stimulatory effect. Other aspects of the reproductive physiology such as enlargement of the cloacal diameter and growth of the oviduct also seem to be controlled primarily by estrogens with little or no additive effect of P. These conclusions were confirmed by injecting egg-laying females with an antiestrogen, tamoxifen, or an antiprogestin, RU38486. Only the former had marked effects on sexual receptivity, cloacal diameter, and oviduct weight. The inhibiting effects of tamoxifen could easily be reversed by injecting females with large doses of estrogen, which demonstrates that tamoxifen acts on an estrogen-dependent mechanism and not through nonspecific effects.

Animals↗

Inhibition of testosterone metabolism in the brain and cloacal gland of the quail by specific inhibitors and antihormones.

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.

Androgen Antagonists↗

[Interaction of androgens and estrogens in the control of reproduction].

Castrated male quail were injected with the synthetic oestrogen, diethylstylbestrol (DES) or the synthetic androgen, methyltrienolone (R 1881) or both compounds simultaneously. Both R 1881 and DES activated male sexual behaviour, inhibited LH and FSH secretion and increased hypothalamic aromatase activity. Additive effects between R 1881 and DES were observed for the induction of brain aromatase and for the inhibition of FSH secretion. As a consequence, mechanisms mediated by androgen and estrogen receptors must be involved in the control of these reproductive characteristics.

Androgens↗

Brain testosterone metabolism during ontogeny in the zebra finch.

The testosterone metabolism in different parts of the brain and in the syrinx was studied by an in vitro radioenzyme assay in male and female zebra finch (Taeniopygia guttata castanotis) at various ages between 1 day post-hatch and adulthood. The 5 beta-reductase appeared in all experiments as the main enzyme involved in testosterone metabolism. Two experiments allowed to determine the kinetic parameters of the 5 beta-reductase. It was shown that the Vmax of the 5 beta-reduction is much higher in very young animals than in adults but that the Km of the enzyme is smaller (higher affinity) in the latter than in the former. A major decrease of 5 beta-reductase activity is observed during the ontogeny of both males and females in all the tissue samples which were studied including the hypothalamus and brain regions containing nuclei of the song system. This high reductase activity which produces behaviorally inactive compounds is likely to play a protective role during the sexual differentiation of the birds and in this way prevent the masculinization in females of the song system by the relatively high levels of circulating androgens.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Testosterone-induced brain aromatase is sexually dimorphic.

Female quail are less sensitive than males to the activating effects of testosterone (T) on behavior. Testosterone induces hypothalamic aromatase activity more efficiently in males than in females (higher Vmax but similar Km). As the conversion of T to estradiol (E2) is important for the activation of male sexual behavior, we propose that the incapacity of the female hypothalamus to produce behaviorally active E2 contributes to their insensitivity to T. The 5 alpha-reductase, which converts T into 5 alpha-dihydrotestosterone (5 alpha-DHT), is induced by T specifically in the anterior hypothalamus of both sexes.

Animals↗

A sexually dimorphic nucleus in the quail preoptic area.

The cytoarchitectural analysis of the preoptic-anterior hypothalamic region of the Japanese quail reveals a sexual dimorphism in the total volume of the medial preoptic nucleus (significantly larger in males than in females). Different nuclei of the region (dorsal preopticus, suprachiasmaticus) do not show any statistically significant difference. The sex-related difference is more consistent comparing the distribution of dark volume. This last is due to a larger number of cells containing high amount of Nissl's substance in male than in female. Present findings represent the first example of sexual dimorphism in the avian hypothalamus.

Animals↗

Diurnal variations of sexual receptivity in the female Japanese quail (Coturnix coturnix japonica).

A series of experiments was performed to study the changes during the day of female receptivity in Japanese quail. In each experiment, the frequency of squatting and the percentage of male approaches which were followed by squatting increased at the end of the day, approximately 11 to 13 hr after lights on (in a photoperiod of 16L:8D). In some cases this increased receptivity was associated with a significant decrease of the long-avoid frequency. Analyses are presented which demonstrate that the increased receptivity at the end of the day is not directly caused by the oviposition and does not result directly from changes in the behavior of the male stimuli. This is strongly supported by the observation that the increase in receptivity was observed at the same time after lights on (but different clock times) in two groups of females which were raised in two different photoperiods shifted by 6 hr and tested with the same group of males raised in one of the two photoperiods. The increase in receptivity coincides with an increase in plasma estradiol and progesterone. Considering that this behavior is suppressed by ovariectomy, it is argued that the daily changes in receptivity could be controlled by the hormonal changes associated with the ovulatory cycle.

Animals↗