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See detailAromatase-Immunoreactive Cells Are Present in Mouse Brain Areas That Are Known to Express High Levels of Aromatase Activity
Foidart, Agnès ULg; Harada, N.; Balthazart, Jacques ULg

in Cell & Tissue Research (1995), 280(3), 561-74

The transformation of testosterone into estradiol in the brain plays a key role in several behavioral and physiological processes, but it has been so far impossible to localize precisely the cells of the ... [more ▼]

The transformation of testosterone into estradiol in the brain plays a key role in several behavioral and physiological processes, but it has been so far impossible to localize precisely the cells of the mammalian brain containing the relevant enzyme, viz., aromatase. We have recently established an immunohistochemical technique that allows the visualization of aromatase-immunoreactive cells in the quail brain. In this species, a marked increase in the optical density of aromatase-immunoreactive cells is observed in subjects that have been treated with the aromatase inhibitor, R76713 or racemic Vorozole. This increased immunoreactivity, associated with a total blockade of aromatase activity, has been used as a tool in the present study in which the distribution of aromatase-immunoreactive material has been reassessed in the brain of mice pretreated with R76713. As expected, the aromatase inhibitor increases the density of the immunoreactive signal in mice. Strongly immunoreactive cells are found in the lateral septal region, the bed nucleus of the stria terminalis, the central amygdala, and the dorso-lateral hypothalamus. A less dense signal is also present in the medial preoptic area, the nucleus accumbens, several hypothalamic nuclei (e.g., paraventricular and ventromedial nuclei), all divisions of the amygdala, and several regions of the cortex, especially the cortex piriformis. These data demonstrate that, contrary to previous claims, aromatase-immunoreactive cells are present in all brain regions that have been shown previously to contain high aromatase activity. [less ▲]

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See detailAppetitive as Well as Consummatory Aspects of Male Sexual Behavior in Quail Are Activated by Androgens and Estrogens
Balthazart, Jacques ULg; Reid, J.; Absil, Philippe ULg et al

in Behavioral Neuroscience (1995), 109(3), 485-501

Appetitive male sexual behavior was measured in male quail with the use of a learned social proximity procedure that quantified the time spent by a male in front of a window providing a view of a female ... [more ▼]

Appetitive male sexual behavior was measured in male quail with the use of a learned social proximity procedure that quantified the time spent by a male in front of a window providing a view of a female that was subsequently released into the cage, providing an opportunity for copulation. The learned response is not acquired by castrated males but can be acquired when castrates are treated with testosterone (T) or with the synthetic estrogen diethylstilbestrol or with the endogenous estrogen 17 beta-estradiol. Only birds that become sexually active acquire the response. Conversely, birds in which the consummatory copulatory behavior is disrupted by treatment with the antiestrogen tamoxifen lose the anticipatory response. These results demonstrate that appetitive sexual behavior is, like copulation, activated by T and by estrogens. This suggests that intracerebral aromatization of T also plays a critical role in the activation of this behavior. [less ▲]

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See detailCritical Re-Examination of the Distribution of Aromatase-Immunoreactive Cells in the Quail Forebrain Using Antibodies Raised against Human Placental Aromatase and against the Recombinant Quail, Mouse or Human Enzyme
Foidart, Agnès ULg; Reid, J.; Absil, Philippe ULg et al

in Journal of Chemical Neuroanatomy (1995), 8(4), 267-82

Mouse and quail aromatase cDNAs were isolated from libraries of mouse ovary and quail brain by using a human aromatase cDNA fragment (hA-24) as a probe. These three cDNAs were inserted into plasmid ... [more ▼]

Mouse and quail aromatase cDNAs were isolated from libraries of mouse ovary and quail brain by using a human aromatase cDNA fragment (hA-24) as a probe. These three cDNAs were inserted into plasmid vectors and expressed in Escherichia coli. Antisera against these purified recombinant proteins were raised in rabbit and purified by ammonium sulfate fractionation and affinity chromatography. The three antibodies directed against recombinant human, mouse and quail proteins were used to visualize aromatase-immunoreactive cells in the quail brain. They were compared with the antibody raised against human placental aromatase used in previous experiments and with another antibody recently developed by similar methods. The signal obtained with all antibodies was completely abolished by preadsorption with the homologous recombinant antigens and the signal produced by the two antibodies raised against placental aromatase was similarly abolished by a preadsorption with recombinant quail aromatase. The antibodies raised against recombinant proteins identified the major groups of aromatase cells previously described in the quail brain. The antibodies directed against the mouse and quail antigen identified more positive cells and stained them more densely than the antibodies raised against human recombinant antigen or purified placental aromatase. The new cell groups identified by the antibody raised against quail recombinant aromatase were located in an area ventral to the fasciculus prosencephali lateralis, the nucleus accumbens, the paleostriatum ventrale, the nucleus taeniae, the area around the nucleus ovoidalis, the caudal tuber and the mesencephalic central gray. A critical re-examination of the distribution and nomenclature of the aromatase-positive cells is proposed based on these new findings. [less ▲]

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See detailLes phéromones humaines: vestige animal ou réalité non reconnue
Foidart, Agnès ULg; Legros, Jean-Jacques ULg; Balthazart, Jacques ULg

in Revue Médicale de Liège (1994), 49(12), 662-80

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See detailEffects of Steroidal and Non Steroidal Aromatase Inhibitors on Sexual Behavior and Aromatase-Immunoreactive Cells and Fibers in the Quail Brain
Foidart, Agnès ULg; Harada, N.; Balthazart, Jacques ULg

in Brain Research (1994), 657(1-2), 105-23

Castrated quail were treated with Silastic implants filled with testosterone (T) in association with injections of the aromatase inhibitors, R76713 (racemic vorozole; 1 mg/kg twice a day) or 4 ... [more ▼]

Castrated quail were treated with Silastic implants filled with testosterone (T) in association with injections of the aromatase inhibitors, R76713 (racemic vorozole; 1 mg/kg twice a day) or 4-hydroxyandrostenedione (OHA; 5 mg/bird twice a day). [less ▲]

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See detailSynergistic Control by Androgens and Estrogens of Aromatase in the Quail Brain
Balthazart, Jacques ULg; Stoop, R.; Foidart, Agnès ULg et al

in Neuroreport (1994), 5(14), 1729-32

Castrated quail were injected with testosterone or with the synthetic hormones diethylstilbestrol (DES) or methyltrienolone (R1881) to analyse the steroid specificity in the induction of brain aromatase ... [more ▼]

Castrated quail were injected with testosterone or with the synthetic hormones diethylstilbestrol (DES) or methyltrienolone (R1881) to analyse the steroid specificity in the induction of brain aromatase. R1881 produced a moderate (generally non-significant) increase in the number of aromatase-immunoreactive cells. DES significantly increased the number of positive cells in most brain areas. A clear synergism between DES and R1881 was observed in all brain regions: more immunoreactive cells were found in birds receiving both compounds than in those injected with DES or R1881 alone. DES and R1881 are highly specific ligands for oestrogen and androgen receptors respectively. It appears likely that both androgens and oestrogens directly modulate brain aromatase, presumably at the transcription level. [less ▲]

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See detailAfferent and Efferent Connections of the Sexually Dimorphic Medial Preoptic Nucleus of the Male Quail Revealed by in Vitro Transport of Dii
Balthazart, Jacques ULg; Dupiereux, V.; Aste, N. et al

in Cell & Tissue Research (1994), 276(3), 455-75

The medial preoptic nucleus of the Japanese quail is a testosterone-sensitive structure that is involved in the control of male copulatory behavior. The full understanding of the role played by this ... [more ▼]

The medial preoptic nucleus of the Japanese quail is a testosterone-sensitive structure that is involved in the control of male copulatory behavior. The full understanding of the role played by this nucleus in the control of reproduction requires the identification of its afferent and efferent connections. In order to identify neural circuits involved in the control of the medial preoptic nucleus, we used the lipophilic fluorescent tracer DiI implanted in aldheyde-fixed tissue. Different strategies of brain dissection and different implantation sites were used to establish and confirm afferent and efferent connections of the nucleus. Anterograde projections reached the tuberal hypothalamus, the area ventralis of Tsai, and the substantia grisea centralis. Dense networks of fluorescent fibers were also seen in several hypothalamic nuclei, such as the anterior medialis hypothalami, the paraventricularis magnocellularis, and the ventromedialis hypothalami. A major projection in the dorsal direction was also observed from the medial preoptic nucleus toward the nucleus septalis lateralis and medialis. Afferents to the nucleus were seen from all these regions. Implantation of DiI into the substantia grisea centralis also revealed massive bidirectional connections with a large number of more caudal mesencephalic and pontine structures. The substantia grisea centralis therefore appears to be an important center connecting anterior levels of the brain to brain-stem nuclei that may be involved in the control of male copulatory behavior. [less ▲]

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See detailTestosterone Effects on Neurotensin-Immunoreactive Cells in the Quail Preoptic Area
Absil, Philippe ULg; Balthazart, Jacques ULg

in Neuroreport (1994), 5(9), 1129-32

In the quail preoptic area (POA), neurotensin-immunoreactive (NT-ir) cells are mostly located in the sexually dimorphic medial preoptic nucleus (POM). They are more numerous in females than in males and ... [more ▼]

In the quail preoptic area (POA), neurotensin-immunoreactive (NT-ir) cells are mostly located in the sexually dimorphic medial preoptic nucleus (POM). They are more numerous in females than in males and, in females, their number varies during the ovulatory cycle. This suggests a control by steroids of NT expression in the POA. This idea was tested by comparing the NT-ir populations in gonadectomized males and females treated or not with testosterone. No sex differences in the NT-ir cell number was present in gonadectomized birds. Testosterone treatment increased the number of immunoreactive perikarya. This effect was anatomically specific and the increase was observed in the area caudal to the anterior commissure but not in POM itself. [less ▲]

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See detailSex Difference in the Neurotensin-Immunoreactive Cell Populations of the Preoptic Area in Quail (Coturnix Japonica)
Absil, Philippe ULg; Balthazart, Jacques ULg

in Cell & Tissue Research (1994), 276(1), 99-116

The distribution of neurotensin-immunoreactive cells and fibers was analyzed by immunocytochemistry in the forebrain of male and female Japanese quail (Coturnix japonica) by using an antibody directed ... [more ▼]

The distribution of neurotensin-immunoreactive cells and fibers was analyzed by immunocytochemistry in the forebrain of male and female Japanese quail (Coturnix japonica) by using an antibody directed against the C-terminal part of the molecule. [less ▲]

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See detailAromatase-Immunoreactive Cells in the Quail Brain: Effects of Testosterone and Sex Dimorphism
Foidart, Agnès ULg; de Clerck, A.; Harada, N. et al

in Physiology & Behavior (1994), 55(3), 453-64

We previously demonstrated that testosterone (T) increases aromatase activity (AA) and that AA is sexually dimorphic (males > females) in the quail preoptic area (POA). The precise anatomical localization ... [more ▼]

We previously demonstrated that testosterone (T) increases aromatase activity (AA) and that AA is sexually dimorphic (males > females) in the quail preoptic area (POA). The precise anatomical localization of these effects is, however, impossible to obtain by biochemical assays even when samples are dissected by the Palkovits punch technique. We were recently able to set up an immunocytochemical (ICC) procedure that permits visualization of aromatase-immunoreactive (ARO-ir) cells in the quail brain. This showed that the ARO-ir cells of the quail POA actually outline the sexually dimorphic medial preoptic nucleus (POM). This ICC technique was used here to analyze the sex dimorphism of the quail preoptic aromatase and the localization of T effects on ARO-ir cells. In Experiment 1, the number of ARO-ir cells was counted in one section every 100 microns throughout the rostral to caudal extent of the POM of castrated birds that had been treated with increasing doses of T (5, 10, or 20 mm long Silastic implants). These T-treatments produced a dose-related increase in the sexual behavior of the birds and they increased the number of ARO-ir cells in POM, in the septal regions, and in the bed nucleus of the stria terminalis (BNST). The effect had a particularly large amplitude in the part of the POM located under the anterior commissure (AC). In Experiment 2, the same procedure was used to reanalyze the sex difference of the preoptic aromatase system. This showed that the POM of adult males contains more stained cells than the POM of females but only in a restricted region located just under and rostral to the AC. No significant sex difference was observed in the septum or in the BNST. In Experiment 3, the number of ARO-ir cells was determined in the POM of males and females that had been gonadectomized and treated with a same dose of T (40 mm implants). No sex difference in the number of ARO-ir cells could be detected in these conditions. This suggests that the sex difference in AA that had been previously observed in T-treated birds results either from a difference in aromatase concentration or activity in a similar number of positive cells or from a difference in the number of ARO-ir cells that is very discrete from the anatomical point of view.(ABSTRACT TRUNCATED AT 400 WORDS) [less ▲]

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See detailEffects of Apomorphine on Sexual Behavior in Male Quail
Absil, Philippe ULg; Das, S.; Balthazart, Jacques ULg

in Pharmacology, Biochemistry & Behavior (1994), 47(1), 77-88

In the rat, dopamine (DA) facilitates male copulatory behavior. Indirect evidence based largely on neuroanatomical data suggest that in quail DA is also implicated in the control of male reproductive ... [more ▼]

In the rat, dopamine (DA) facilitates male copulatory behavior. Indirect evidence based largely on neuroanatomical data suggest that in quail DA is also implicated in the control of male reproductive behavior but there is no pharmacological evidence to support this conclusion. To test this idea, castrated testosterone (T)-treated male quail were injected with various doses of the dopaminergic agonist apomorphine (APO) in the range 1-10,000 micrograms/kg. The sexual behavior of birds was recorded starting 15 min after APO injection for a duration of 30 min. A dose-dependent inhibition of male reproductive behavior that lasted for the entire duration of the test was observed. In a second experiment, gonadectomized T-treated male Japanese quail were injected daily with APO (0, 10, or 1,000 micrograms/kg) during 8 days. Their sexual interactions with a partner were quantified either 24 h or 15 min after the last injection. No influence of the treatment on copulatory behavior was observed 24 h after the last injection, but a strong inhibition was present when the test was performed 15 min after. To research whether the inhibitory effects of APO were due to a preferential action on D2 presynaptic autoreceptors, male quail were pretreated with two different D2 antagonists (spiperone or pimozide; 0.5 or 2 mg/kg) before being injected with APO (100 micrograms or 1 mg/kg). Spiperone facilitated male sexual behavior but did not suppress the inhibitory effect of APO. No significant effect of pimozide was observed. These results support the notion that DA modulates male sexual activity in the Japanese quail. The specific role of the different dopaminergic receptor subtypes remains, however, to be elucidated. [less ▲]

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See detailDistribution and Regulation of Estrogen-2-Hydroxylase in the Quail Brain
Balthazart, Jacques ULg; Stoop, R.; Foidart, Agnès ULg et al

in Brain Research Bulletin (1994), 35(4), 339-45

The anatomical distribution and endocrine regulation of the estrogen-2-hydroxylase activity were investigated in the brain of adult male and female Japanese quail. Significant levels of enzymatic activity ... [more ▼]

The anatomical distribution and endocrine regulation of the estrogen-2-hydroxylase activity were investigated in the brain of adult male and female Japanese quail. Significant levels of enzymatic activity were detected in all brain regions that were studied, but the highest levels were observed in preoptic and hypothalamic brain nuclei that are known to contain high levels of aromatase activity. These data are consistent with previous results suggesting that the placental aromatase is also responsible for the estrogen-2-hydroxylase activity. However, there is a marked sex difference and a control by T of aromatase activity in the quail brain, and no such difference in 2-hydroxylase activity could generally be detected except in the VMN. Further studies will be needed to know whether the previously published conclusions concerning the human placenta also apply to the brain. The present data are consistent with the idea that estrogens formed locally in the brain by testosterone aromatization could affect reproduction by interfering with the catecholaminergic transmission after being metabolized into catechol-estrogens. [less ▲]

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See detailThe Catecholaminergic System of the Quail Brain: Immunocytochemical Studies of Dopamine Beta-Hydroxylase and Tyrosine Hydroxylase
Bailhache, T.; Balthazart, Jacques ULg

in Journal of Comparative Neurology (The) (1993), 329(2), 230-56

The distribution of dopamine beta-hydroxylase and tyrosine hydroxylase, two key enzymes in the biosynthesis of catecholamines, was investigated by immunocytochemistry in the brain of male and female ... [more ▼]

The distribution of dopamine beta-hydroxylase and tyrosine hydroxylase, two key enzymes in the biosynthesis of catecholamines, was investigated by immunocytochemistry in the brain of male and female Japanese quail. Cells or fibers showing dopamine beta-hydroxylase and tyrosine hydroxylase immunoreactivity were considered to be noradrenergic or adrenergic, while all structures showing only tyrosine hydroxylase immunoreactivity were tentatively considered to be dopaminergic. The major dopaminergic and noradrenergic cell groups that have been identified in the brain of mammals could be observed in the Japanese quail, with the exception of a tuberoinfundibular dopaminergic group. The dopamine beta-hydroxylase-immunoreactive cells were found exclusively in the pons (locus ceruleus and nucleus subceruleus ventralis) and in the medulla (area of the nucleus reticularis). The tyrosine hydroxylase-immunoreactive cells had a much wider distribution and extended from the preoptic area to the level of the medulla. They were, however, present in larger numbers in the area ventralis of Tsai and in the nucleus tegmenti pedunculo-pontinus, pars compacta, which respectively correspond to the ventral tegmental area and to the substantia nigra of mammals. A high density of dopamine beta-hydroxylase- and tyrosine hydroxylase-immunoreactive fibers and punctate structures was found in several steroid-sensitive brain regions that are implicated in the control of reproduction. In the preoptic area and in the region of the nucleus accumbens-nucleus stria terminalis, immunonegative perikarya were completely surrounded by immunoreactive fibers forming basket-like structures. Given that some of these cells contain the enzyme aromatase, these structures may represent the morphological substrate for a regulation of aromatase activity by catecholamines. The dopamine beta-hydroxylase-immunoreactive fibers were also present in a larger part of the preoptic area of females than in males. This sex difference in the noradrenergic innervation of the preoptic area presumably reflects the sex difference in norepinephrine content in this region. [less ▲]

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See detailBrain Aromatase and the Control of Male Sexual Behavior
Balthazart, Jacques ULg; Foidart, Agnès ULg

in Journal of Steroid Biochemistry & Molecular Biology (1993), 44(4-6), 521-40

The activational effects of testosterone (T) on male copulatory behavior are mediated by its aromatization into estradiol. In quail, we have shown by stereotaxic implantation of steroids and metabolism ... [more ▼]

The activational effects of testosterone (T) on male copulatory behavior are mediated by its aromatization into estradiol. In quail, we have shown by stereotaxic implantation of steroids and metabolism inhibitors and by electrolytic lesions that the action of T and its aromatization take place in the sexually dimorphic medial preoptic nucleus (POM). The distribution and regulation of brain aromatase was studied in this species by product-formation assays measuring aromatase activity (AA) in microdissected brain regions and by immunocytochemistry (ICC). Aromatase-immunoreactive (ARO-ir) neurons were found in four brain regions: the POM, the septal region, the bed nucleus of the stria terminals (BNST) and the tuberal hypothalamus. ARO-ir cells actually outline the POM boundaries. ARO-ir material is found not only in the perikarya of neurons but also in the full extension of their cellular processes including the axons and the presynaptic boutons. This is confirmed at the light level by the demonstration of immunoreactive fibers and punctate structures in brain regions that are sometimes fairly distant from the closest ARO-ir cells. A lot of ARO-ir cells in the POM and BNST do not contain immunoreactive estrogen receptors (ER-ir) as demonstrated by double label ICC. These morphological data suggest an unorthodox role for the enzyme or the locally formed estrogens. In parallel with copulatory behavior, the preoptic AA decreases after castration and is restored by T to levels seen in sexually mature males. This probably reflects a change in enzyme concentration rather than a modulation of the activity in a constant number of molecules since the maximum enzymatic velocity (Vmax) only is affected while the affinity (Km) remains unchanged. In addition, T increases the number of ARO-ir neurons in POM and other brain areas suggesting that the concentration of the antigen is actually increased. This probably involves the direct activation of aromatase transcription as demonstrated by RT-PCR studies showing that aromatase mRNA is increased following T treatment of castrates. These activating effects of T seem to result from a synergistic action of androgenic and estrogenic metabolites of the steroid. The anatomical substrate for these regulations remains unclear at present especially in POM where ARO-ir cells do not in general contain ER-ir while androgen receptors appear to be rare based on both [3H] dihydrotestosterone autoradiography and ICC. Transynaptic mechanisms of control may be considered. A modulation of brain aromatase by catecholamines is also suggested by a few pharmacological studies.(ABSTRACT TRUNCATED AT 400 WORDS) [less ▲]

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See detailEffects of Neurochemical Lesions of the Preoptic Area on Male Sexual Behavior in the Japanese Quail
Bailhache, T.; Surlemont, C.; Balthazart, Jacques ULg

in Brain Research Bulletin (1993), 32(3), 273-83

Two experiments were carried out during which the noradrenergic neurotoxin, 5-amino-2,4-dihydroxy-alpha-methylphenylethylamine (5-ADMP) was applied to the brain of quail in order to evaluate the role of ... [more ▼]

Two experiments were carried out during which the noradrenergic neurotoxin, 5-amino-2,4-dihydroxy-alpha-methylphenylethylamine (5-ADMP) was applied to the brain of quail in order to evaluate the role of the noradrenergic system in the control of male copulatory behavior. In the first experiment, the ICV injection of 5-ADMP slightly enhanced the sexual behavior observed in testosterone (T)-treated castrated male quail. This brings additional support to the notion that norepinephrine tonically inhibits male copulatory behavior in quail. In the second experiment, 5-ADMP implanted directly into the preoptic area disrupted the restoration by T of copulatory behavior in castrated quail and, at the same time, produced a brain lesion that partly destroyed the sexually dimorphic medial preoptic nucleus, a previously established site of T action on behavior. These lesions produced by a high (presumably too high) concentration of neurotoxin provided an independent confirmation of effects previously observed after electrolytic lesions. Correlation analyses also confirmed that the medial part of the POM just rostral to the anterior commissure is more closely associated with copulatory behavior and may, therefore, represent a key center for steroid action on this behavior. [less ▲]

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See detailBehavioral Demasculinization of Female Quail Is Induced by Estrogens: Studies with the New Aromatase Inhibitor, R76713
Balthazart, Jacques ULg; De Clerck, A.; Foidart, Agnès ULg

in Hormones & Behavior (1992), 26(2), 179-203

The injection before Day 12 of incubation of estradiol benzoate (EB) into Japanese quail eggs produces a complete behavioral demasculinization of adult males that will hatch from these eggs. These males ... [more ▼]

The injection before Day 12 of incubation of estradiol benzoate (EB) into Japanese quail eggs produces a complete behavioral demasculinization of adult males that will hatch from these eggs. These males never show copulatory behavior even after administration of high levels of exogenous testosterone (T). It is usually assumed that such a demasculinization normally takes place in female embryos under the influence of endogenous estrogens but few experimental data are available to confirm the validity of this model. A series of four experiments was performed during which R76713, a triazole derivative that specifically inhibits aromatase (estrogen synthetase) activity, was injected into quail eggs at different stages of incubation to prevent the production of endogenous estrogens. The consequences of these embryonic treatments on the T-activated sexual behavior in adults were then quantified. When injected before Day 12 of incubation, R76713 completely blocked the behavioral demasculinization of females without affecting the behavior of the males. After a treatment with T, almost all R76713-treated females showed as adults a masculine copulatory behavior that was undistinguishable from the behavior of intact males. This effect was fully reversed by the injection in egg of EB demonstrating that the effects of R76713 were specifically due to the suppression of endogenous estrogens. Injection of R76713 during the late phase of the incubation (Day 12 or Day 15) only maintained weak copulatory behavior in females which confirmed that the behavioral demasculinization in quail takes place mainly though not exclusively during the early stages of ontogeny. In a last experiment, we combined an early R76713 treatment with an injection of EB either on Day 9 or on Day 14 of incubation. This showed that the sensitivity to differentiating effects of estrogens varies with age in a sexually differentiated manner. The EB injection on Day 9 demasculinized both male and female embryos. If this injection was delayed until Day 14, it was no longer effective in males but still caused a partial demasculinization of females. This demonstrates that even if females are not yet behaviorally demasculinized on Day 9 of incubation (suppression of aromatase activity at that age will maintain the behavior), their sensitivity to estrogens is already different from that of males. [less ▲]

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See detailImmunocytochemical Localization of Androgen Receptors in the Male Songbird and Quail Brain
Balthazart, Jacques ULg; Foidart, Agnès ULg; Wilson, E. M. et al

in Journal of Comparative Neurology (The) (1992), 317(4), 407-20

The distribution of androgen receptors was studied in the brain of the Japanese quail (Coturnix japonica), the zebra finch (Taeniopygia guttata), and the canary (Serinus canaria) by immunocytochemistry ... [more ▼]

The distribution of androgen receptors was studied in the brain of the Japanese quail (Coturnix japonica), the zebra finch (Taeniopygia guttata), and the canary (Serinus canaria) by immunocytochemistry with a polyclonal antibody (AR32) raised in rabbit against a synthetic peptide corresponding to a sequence located at the N-terminus of the androgen receptor molecule. In quail, androgen receptor-immunoreactive cells were observed in the nucleus intercollicularis and in various nuclei of the preoptic-hypothalamic complex, namely, the nucleus preopticus medialis, the ventral part of the nucleus anterior medialis hypothalami, the nucleus paraventricularis magnocellularis, the nucleus ventromedialis hypothalami, and the tuberal hypothalamus. In the two songbird species, labeled cells were also observed in various nuclei in the preoptic-hypothalamic region, in the nucleus taeniae, and in the nucleus intercollicularis. Additional androgen receptor-immunoreactive cells were present in the androgen-sensitive telencephalic nuclei that are part of the song control system. These immunoreactive cells filled and outlined the boundaries of the hyperstriatum ventrale, pars caudalis, nucleus magnocellularis neostriatalis anterioris (both in the lateral and medial subdivisions), and nucleus robustus archistriatalis. The immunoreactive material was primarily present in cell nuclei but a low level of immunoreactivity was also clearly detected in cytoplasm in some brain areas. These studies demonstrate, for the first time, that androgen receptors can be detected by immunocytochemistry in the avian brain and the results are in general agreement with the binding data obtained by autoradiography with tritiated dihydrotestosterone. Immunocytochemical methods offer several advantages over autoradiography and their use for the study of the androgen receptor will greatly facilitate the analysis of steroid-sensitive systems in the avian brain. [less ▲]

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See detailNeuroanatomical Specificity in the Autoregulation of Aromatase-Immunoreactive Neurons by Androgens and Estrogens: An Immunocytochemical Study
Balthazart, Jacques ULg; Foidart, Agnès ULg; Surlemont, C. et al

in Brain Research (1992), 574(1-2), 280-90

Testosterone (T) increases brain aromatase activity (AA) in quail and other avian and mammalian species. It was shown both in quail and in rat that this enzymatic induction results from a synergistic ... [more ▼]

Testosterone (T) increases brain aromatase activity (AA) in quail and other avian and mammalian species. It was shown both in quail and in rat that this enzymatic induction results from a synergistic action of androgens and estrogens. These studies provide little information on possible anatomical or cellular specificity of the effect. Using a polyclonal antiserum against human placental aromatase, we have previously identified aromatase-immunoreactive (ARO-ir) neurons in the quail brain and demonstrated that T increases the number of ARO-ir cells in the quail preoptic area (POA) supporting previous evidence that T increases AA in the brain. However, which T metabolites are involved, the actual mechanism of regulation and the possibility of anatomical specificity for these effects are not yet clear. In the present study, we disassociated the effects of androgens and estrogens in aromatase induction by comparing ARO-ir neurons of quail treated with T alone or T in the presence of a potent aromatase inhibitor (R76713), which has been shown to depress AA levels and to suppress T-activated copulatory behavior. T increased the number of ARO-ir cells in POA, bed nucleus striae terminalis (BNST) and tuberal hypothalamus (Tu). The T effect was inhibited by concurrent treatment with aromatase inhibitor in Tu, but not in POA and BNST. This differential effect of the aromatase inhibitor fits in very well with our previous studies of the co-localization of aromatase and estrogen receptors. The T effect was blocked by R76713 in areas where ARO-ir and estrogen receptor-ir are generally co-localized (Tu) and was not affected in areas with mainly ARO-ir positive, estrogen receptor-ir negative cells (POA, BNST). This suggests anatomical differences in the expression or clearance of aromatase which may be differentially sensitive to androgens and estrogens and dependent upon the presence of sex steroid receptors. [less ▲]

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See detailAromatase as a Cellular Marker of Testosterone Action in the Preoptic Area
Balthazart, Jacques ULg; Surlemont, C.; Harada, N.

in Physiology & Behavior (1992), 51(2), 395-409

We recently showed, using a new immunocytochemical technique, that aromatase-immunoreactive neurons are a specific marker for the sexually dimorphic medial preoptic nucleus (POM) in quail and that the ... [more ▼]

We recently showed, using a new immunocytochemical technique, that aromatase-immunoreactive neurons are a specific marker for the sexually dimorphic medial preoptic nucleus (POM) in quail and that the number of these immunoreactive cells is markedly increased by a systemic treatment with testosterone (T). Since the POM is a key site for the activation of copulatory behavior by T and this androgen must be converted into estrogen by local aromatization within the POM before it can exert its behavioral effects, we used aromatase immunocytochemistry to map, at a cellular level of resolution, the areas that are destroyed by electrolytic lesions or that are stimulated by the stereotaxic implantation of T in the preoptic area (POA). These measures of the cellular action of T in the preoptic area were then correlated with the behavior of the animals to identify the parts of the POA that are critical in the activation of behavior. The electrolytic lesions of the POA disrupted the activation of male sexual behavior by T only if they destroyed a significant part of the POM. All lesions reduced the volume of the dimorphic nucleus and the absolute number of its aromatase-immunoreactive neurons, but the density of these cells in the remaining POM was not affected, suggesting that the volume change in the nucleus reflected a centripetal displacement of its boundaries rather than an overall shrinkage of the structure. Stereotaxic T implants in or close to POM activated male copulatory behavior and increased the volume of the POM and the number of its aromatase-immunoreactive cells. These neuroanatomical effects were more prominent on the side of the implant, but they were also detected on the contralateral side. Correlative analyses suggested that a part of the POM just rostral to the anterior commissure is critical for the activation of copulatory behavior. The best correlations between the behavioral deficits induced by electrolytic lesions and the size of the lesions were indeed observed in this area. In addition, high correlations were also observed between the behavior activated by T implants and the POM size or number of aromatase-immunoreactive cells that were induced by T in this area. Aromatase immunocytochemistry therefore appears as a useful tool to map the brain areas in which T action is presumably critical for the activation of male sexual behavior. It has allowed us to identify in the present studies a small part of the sexually dimorphic POM that is closely associated with behavior.(ABSTRACT TRUNCATED AT 400 WORDS) [less ▲]

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See detailEffects of Alpha-Methyl-Para-Tyrosine on Monoamine Levels in the Japanese Quail: Sex Differences and Testosterone Effects
Balthazart, Jacques ULg; Foidart, Agnès ULg; Sante, P. et al

in Brain Research Bulletin (1992), 28(2), 275-88

Experiments were performed to obtain more information on the regulation by steroids of catecholaminergic systems in the brain of Japanese quail. Dose-response and time-response experiments were first ... [more ▼]

Experiments were performed to obtain more information on the regulation by steroids of catecholaminergic systems in the brain of Japanese quail. Dose-response and time-response experiments were first performed to determine optimal conditions for measuring turnover in the quail brain. The norepinephrine and dopamine turnover were then estimated in microdissected brain nuclei of birds that were either sexually mature or gonadectomized or gonadectomized and treated with testosterone. Two major facts that bear direct relationship with the control of masculine reproductive behavior were demonstrated. On one hand, the dopamine turnover in the medial preoptic nucleus, a sexually dimorphic brain structure which is critically implicated in the control of copulatory behavior was much higher in male than in female quail irrespective of the hormonal condition of the birds. On the other hand, norepinephrine concentrations appeared to be higher in several nuclei of the female brain by comparison with males. These sex differences might represent part of the causal factors that underlie the sex dimorphism in reproductive behavior in quail. [less ▲]

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