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J Balthazart

Publications and source records attributed to J Balthazart.

At least 109 records · Page 6Linked to original sources
PubMed1995

Pre- and post-translational regulation of aromatase by steroidal and non-steroidal aromatase inhibitors.

Treatment of castrated quail with testosterone (T) reliably activates male copulatory behavior and, at the same time, increases the aromatase activity (AA), the number of aromatase-immunoreactive (ARO-ir) cells and the concentration of aromatase mRNA as measured by RT-PCR in the brain. All these effects can be mimicked by estrogens. The behavioral effects of T can be blocked by a variety of aromatase inhibitors and, in parallel, the AA is strongly inhibited in the preoptic area (POA). We showed recently that the steroidal inhibitor, 4-OH-androstenedione (OHA) markedly decreases the immunostaining density of brain ARO-ir cells while the non-steroidal inhibitor, R76713 (racemic Vorozole; VOR) unexpectedly increased the density of this staining, despite the fact that the enzyme activity was completely inhibited. To generalize these findings and try to identify the underlying mechanism, we compared here the effects of two steroidal (OHA and androstatrienedione [ATD]) and two non-steroidal (VOR and Fadrozole [FAD]) aromatase inhibitors on the aromatase immunostaining and aromatase mRNA concentration in the brain of castrated quail concurrently treated with T. The 4 inhibitors significantly blocked the activation by T of male copulation. The two steroidal inhibitors decreased the immunostaining of brain ARO-ir cells but both VOR and FAD markedly enhanced the density of this staining. In parallel, OHA and ATD completely blocked the T-induced increase in aromatase mRNA concentration, while VOR and FAD had no effect on these RNA concentrations in the POA-anterior hypothalamus and they decreased them only slightly in the posterior hypothalamus. Taken together these results suggest that the inhibition of AA by ATD or OHA and the subsequent removal of locally produced estrogens blocks the synthesis of aromatase presumably at the transcriptional level. By contrast, the two non-steroidal inhibitors tested here block AA but in parallel increase the aromatase immunostaining. This effect does not result from an enhanced transcription and it is therefore speculated that these compounds increase either the translation of the aromatase mRNA or the half-life of the protein itself.

Animals↗
PubMed1995

Assessment of volumetric sex differences in the song control nuclei HVC and RA in zebra finches by immunocytochemistry for methionine enkephalin and vasoactive intestinal polypeptide.

In this study we assessed sex differences in the volume of two vocal control nuclei, the high vocal center (HVC) and the robust nucleus of the archistriatum (RA) in zebra finches (Taeniopygia guttata) exposed to various endocrine treatments (neonatal treatment with an aromatase inhibitor and/or an adult treatment with testosterone). The boundaries of these nuclei were defined in sections stained for Nissl substance and compared to alternate sections stained with an immunocytochemical procedure for two neuropeptides, methionine enkephalin and vasoactive intestinal polypeptide. A high density of immunoreactive fibers for methionine enkephalin and vasoactive intestinal polypeptide completely covered the high vocal center in male and female zebra finches and these fibers were not observed in the surrounding neostriatum. With the use of these markers to define the boundary of the high vocal center, it became possible to reconstruct its volume. Positively staining perikarya were not apparent within the boundaries of this nucleus. Immunoreactive fibers for vasoactive intestinal polypeptide and enkephalin also allowed one to define the boundary of the robust nucleus of the archistriatum but they did not fill the entire area of the nucleus as is the case for the high vocal center. When the volumes of both nuclei were reconstructed based on enkephalin or vasoactive intestinal polypeptide immunoreactivity, the presence of a marked sex difference in the volume of these nuclei was confirmed. Neonatal and adult endocrine manipulations did not affect the volumes of these two nuclei measured in Nissl-stained material nor did they affect the volumes as defined based on the peptidergic innervations of the nuclei. Also, the volumetric estimates of these two nuclei derived from sections stained by immunocytochemistry were in good agreement in all groups of birds with the values obtained based on an analysis of the Nissl-stained material. These results illustrate the usefulness of employing a variety of histochemical markers to define a brain area when investigating brain variation and plasticity.

Animals↗
PubMed1995

Peptidergic delineations of nucleus interface reveal a sex difference in volume.

Nucleus interface (Nif) is a song bird vocal control nucleus located in the neostriatum. In zebra finches (Taeniopygia guttata), high densities of fibers immunoreactive for the vasoactive intestinal polypeptide (VIP) and methionine enkephalin (ENK) define the boundaries of this nucleus. The volume of Nif measured using this pattern of peptidergic immunoreactivity was greater in males than in females. This difference was also found in gonadectomized, testosterone-treated birds that had similar levels of circulating steroids, suggesting that it is organizational in nature. These data show that Nif may form part of the neural substrate underlying sex differences in song behavior. They also provide further evidence that VIP and ENK may play a role in the activation of bird song.

Animals↗
PubMed1995

Aromatase-immunoreactive cells are present in mouse brain areas that are known to express high levels of aromatase activity.

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.

Animals↗
PubMed1995

Critical 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.

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.

Animals↗
PubMed1995

Autoradiographic localization of D1-like dopamine receptors in the forebrain of male and female Japanese quail and their relationship with immunoreactive tyrosine hydroxylase.

The distribution of D1-like dopamine receptors was studied in the brain of male and female Japanese quail (Coturnix japonica) by means of quantitative autoradiography with 3H-labelled D1 selective antagonist, SCH 23390, serving as a ligand. A specific, saturable, high affinity binding of this ligand was demonstrated. High densities of binding sites were detected in the lobus parolfactorius, olfactory tubercle, and paleostriatum augmentatum. Medium densities were observed in the entire neostriatum and in the external layers of the optic tectum. Similar levels of binding outlined the paleostriatum primitivum, the nucleus pretecalis and the nucleus intercollicularis. Low but significant levels of receptors were also present in the medial preoptic area at the level of the sexually dimorphic medial preoptic nucleus and throughout the infundibulum, as well as in the ectostriatum, medial and lateral septum, and nucleus accumbens. At the level of the medial septum, just dorsal to the anterior commissure, two circular areas of high receptor density corresponding to the nucleus of the septal commissure were also observed. No sex difference in receptor density could be detected in any of the areas. All areas containing high densities of D1 receptors also contained high densities of tyrosine hydroxylase (TH) fibers. However, certain areas characterized by a high density of TH-immunoreactive fibers did not contain appreciable densities of D1-like dopamine receptors. The distribution of this receptor and its relationship to TH-immunoreactivity is consistent with observations made in other vertebrates, suggesting that the dopaminergic system is evolutionarily highly conserved among amniote vertebrates.

Animals↗
PubMed1995

Sexual differentiation of brain and behavior in quail and zebra finches: studies with a new aromatase inhibitor, R76713.

In many species of vertebrates, major sex differences affect reproductive behavior and endocrinology. Most of these differences do not result from a direct genomic action but develop following early exposure to a sexually differentiated endocrine milieu. In rodents, the female reproductive phenotype mostly develops in the absence of early steroid influence and male differentiation is imposed by the early action of testosterone, acting at least in part through its central conversion into estrogens or aromatization. This pattern of differentiation does not seem to be applicable to avian species. In Japanese quail (Coturnix japonica), injection of estrogens into male embryos causes a permanent loss of the capacity to display male-type copulatory behavior when exposed to testosterone in adulthood. Based on this experimental result, it was proposed that the male reproductive phenotype is "neutral" in birds (i.e. develops in the absence of endocrine influence) and that endogenous estradiol secreted by the ovary of the female embryo is responsible for the physiological demasculinization of females. This model could be recently confirmed. Females indeed display a higher level of circulating estrogens that males during the second part of their embyronic life. In addition, treatment of female embryos with the potent aromatase inhibitor, R76713 or racemic vorozole which suppresses the endogenous secretion of estrogens maintains in females the capacity to display the full range of male copulatory behaviors. The brain mechanisms that control this sexually differentiated behavior have not been identified so far but recent data suggest that they should primarily concern a sub-population of aromatase-immunoreactive neurons located in the lateral parts of the sexually dimorphic preoptic nucleus. The zebra finch (Taeniopygia guttata) exhibits a more complex, still partly unexplained, differentiation pattern. In this species, early treatment with exogenous estrogens produces a masculinization of singing behavior in females and a demasculinization of copulatory behavior in males.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

Appetitive as well as consummatory aspects of male sexual behavior in quail are activated by androgens and estrogens.

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.

Androgens↗
PubMed1994

Distribution of beta-endorphin-like-immunoreactive structures in the chicken and quail brain as demonstrated with a new homologous antibody directed against a synthetic peptide.

A polyclonal rabbit antibody was raised against a synthetic peptide fragment located at the C-terminal end of turkey beta-endorphin (beta-END) and used to analyze the distribution of beta-END-immunoreactive-like structures in the quail and chicken brain. Three major groups of immunopositive cells were detected in the preoptic area-hypothalamus complex. A thin layer of immunopositive cells was parallel and adjacent to the ventral edge of the brain in the preoptic and anterior hypothalamic region, a more numerous group of immunoreactive perikarya was located along the walls of the third ventricle in these same regions, and, finally, a few scattered cells were found in a more lateral position on both the internal and external sides of the tip of the fasciculus prosencephali lateralis. The periventricular cell population extended in the caudal direction until the posterior hypothalamus. Labelled fibers were always associated with these immunoreactive perikarya, and they were also found in the adjacent hypothalamic regions. A dense innervation of the median eminence was also detected. These data are compared with previous studies in mammals and birds that had identified more restricted populations of immunoreactive cells and the possible sources of the observed discrepancy are discussed. The functional significance of the present data is also briefly analyzed.

Animals↗
PubMed1994

Effects of steroidal and non steroidal aromatase inhibitors on sexual behavior and aromatase-immunoreactive cells and fibers in the quail brain.

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). Control birds received no treatment (CX group) or were implanted with T capsules only (CX + T group). Both R76713 and OHA strongly inhibited the T-activated male copulatory behavior. This inhibition had the same magnitude in both groups. The growth of the cloacal gland, a strictly androgen-dependent process was not affected by these compounds. The treatments significantly affected the number of aromatase-immunoreactive (ARO-ir) cells in each of the six brain areas that were studied: the anterior and posterior parts of the sexually dimorphic medial preoptic nucleus (POM), the septal region, the bed nucleus of the stria terminalis (BNST) and the anterior and posterior parts of the tuber. This number was significantly increased in all areas by T. In agreement with our previous study, R76713 significantly inhibited this effect of T in the tuberal hypothalamus but not in the anterior POM nor in the BNST. By contrast the effect of T on the number of ARO-ir cells was completely blocked by OHA in all brain nuclei. The two inhibitors had statistically different effects in all brain regions. Like in a previous study, R76713 increased the intensity of the staining of all ARO-ir cells. This effect took several days to develop suggesting a progressive build-up of the enzyme concentration. This was also suggested by the fact that a rebound in aromatase activity was observed 16 to 24 h after a single injection of R76713. The increased immunoreactivity was not observed in OHA-treated birds. The denser immunoreactivity in R76713-treated birds and the better tissue preservation due to the aldehyde fixative that had been used provided here a clearer picture of the cellular and subcellular localization of ARO-ir material. This allowed to identify new groups of immunoreactive cells, namely in the nucleus accumbens, in the area of the paleostriatum ventrale, in the nucleus taeniae, in the medial and caudal hypothalamus and in the medial part of the mesencephalon and of the pons. Most of the immunoreactive material was located in perikarya but some of these cells were also surrounded by dense networks of ARO-ir fibers often associated with immunopositive punctate structures.(ABSTRACT TRUNCATED AT 400 WORDS)

Androstenedione↗
PubMed1994

Vasotocinergic innervation of sexually dimorphic medial preoptic nucleus of the male Japanese quail: influence of testosterone.

The distribution of vasotocin (VT)-immunoreactive (IR) fibers was described in the preoptic and septal regions of the male quail brain. The density of VT-IR fibers was measured in the sexually dimorphic preoptic nucleus (POM) and lateral septum (SL) of adult male quail (Coturnix japonica) by means of quantitative image analysis. Experimental manipulations of the hormonal environment in the peripubertal period influenced this distribution. In both regions, the VT immunoreactivity was reduced or absent when males were castrated. The immunoreactivity was restored to its original level in castrated males by Silastic implants of testosterone. These changes were anatomically specific as evidenced by the fact that the density of VT fibers did not vary in the hypothalamo-neurohypohysial tract as a function of the endocrine condition of the subjects. No change was also observed in the number of VT-IR cells in the periventricular region close to the POM. Previously published data show that VT or its mammalian homolog, vasopressin are implicated in the control of a wide range of instinctive behaviors. The steroid-dependent VT afferents to the POM, a key area controlling male copulatory behavior in quail could therefore be involved in the control of the sexual behavior in this species. The outputs of the POM which contains steroid-receptors could therefore be modulated by steroids in two different ways: directly through the steroid receptors it contains and indirectly through its steroid-sensitive peptidergic afferents.

Animals↗
PubMed1994

Synergistic control by androgens and estrogens of aromatase in the quail brain.

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.

Androgens↗
PubMed1994

Testosterone effects on neurotensin-immunoreactive cells in the quail preoptic area.

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.

Animals↗
PubMed1994

Morphometric studies demonstrate that aromatase-immunoreactive cells are the main target of androgens and estrogens in the quail medial preoptic nucleus.

The volume and cytoarchitectonic organization of the sexually dimorphic medial preoptic nucleus (POM) of the quail are sensitive to plasma levels of testosterone (T). We previously showed that, in castrated quail, T or its estrogenic metabolite, estradiol (E2), increases the size of the large neurons located in the lateral part of POM. Embryonic treatments with estrogens are also known to affect permanently the size of these large neurons. Since the lateral POM also contains a dense population of aromatase-immunoreactive (ARO-ir) cells, and these are known to be a target for steroids, we hypothesized that the effects of steroids identified in previous experiments were primarily directed to these ARO-ir cells. This idea was tested in two experiments in which the size of these cells was measured in male quail under various endocrine conditions. In experiment 1, a detailed analysis of ARO-ir and of non-immunoreactive cells in the POM of adult, sexually mature males revealed that the immunoreactive perikarya are larger than the non-immunoreactive cells and that they constitute the vast majority of the large cells (area > 50 microns 2) in the POM. In experiment 2, it was shown that T and E2 actually increase the size of ARO-ir cells in the POM while the androgenic metabolite of T, dihydrotestosterone has no effect at this level. Taken together, these data suggest that the sex differences and the steroid-induced changes in cell size previously described in the study of POM sections stained for Nissl material largely concern aromatase-containing cells. Since aromatization of T plays a limiting role in the activation of male copulatory behavior, these changes may represent the morphological signature of the mechanisms causally involved in the control of this behavior.

Androgens↗
PubMed1994

Afferent and efferent connections of the sexually dimorphic medial preoptic nucleus of the male quail revealed by in vitro transport of DiI.

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.

Afferent Pathways↗
PubMed1994

Sex difference in the neurotensin-immunoreactive cell populations of the preoptic area in quail (Coturnix japonica).

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. Immunoreactive perikarya were located almost exclusively in the medial preoptic area with small populations also being present in the nucleus paraventricularis and in the tuberal region. Immunoreactive fibers were observed not only throughout the preoptic area-hypothalamus, but also in the septal region, nucleus intercollicularis, substantia grisea centralis and the classical catecholaminergic areas of the mesencephalon, such as the area ventralis of Tsai and the nucleus tegmenti pedunculo-pontinus, pars compacta. The preoptic neurotensin-immunoreactive cells were exclusively located within the boundaries of the sexually dimorphic medial preoptic nucleus. They were significantly more numerous in females than in males. In females, the number of neurotensin cells varied during the ovulatory cycle: fewer cells were observed in birds that were about to lay an egg (they had a calcified egg in the oviduct) than in those that had already laid or were not going to lay on that day. These data indicate major variations in the expression of neurotensin in response to neurochemical or neuroendocrine changes associated with ovulation.

Animals↗
PubMed1994

Aromatase-immunoreactive cells in the quail brain: effects of testosterone and sex dimorphism.

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)

Animals↗