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Biomedical subjects

B A Schlinger

Publications and source records attributed to B A Schlinger.

At least 37 records · Page 2Linked to original sources
PubMed1998

The passerine hippocampus is a site of high aromatase: inter- and intraspecies comparisons.

The vertebrate hippocampus (HP) plays a critical role in the organization of memory. Estrogens alter synaptic morphology and function in the mammalian HP and may potentiate memory performance. Previous studies suggest that the songbird HP itself is a site of significant aromatase expression, intimating that local estrogen synthesis may provide a source of this steroid to estrogen-sensitive neural circuits. To explore the potential role of local estrogen synthesis on HP structure and function, we have characterized aromatase message and activity in the zebra finch HP. Toward this end we have compared (a) HP aromatase mRNA with that at other neural loci, (b) HP aromatase activity between adults of both sexes, and (c) HP and hypothalamic preoptic area (HPOA) aromatase activity among songbirds and nonsongbirds. Finally we asked whether aromatase activity was intrinsic to the HP by maintaining it in culture, isolated from the rest of the telencephalon. The HP of every songbird studied expresses aromatase, with comparable levels across sexes. Notably, aromatase activity was found at higher levels in the songbird HP than in the HPOA. In both nonsongbird species investigated, however, HP aromatase activity was undetectable under identical assay conditions. Additionally, the developing songbird HP continues to express aromatase when cultured in isolation from the rest of the telencephalon. The data suggest that HP aromatase is characteristic of passeriformes and, as in the HPOA, may represent a mechanism whereby estrogen is made available to neural circuits. Passerines may prove invaluable animal models for investigations of the estrogenic modulation of HP structure and function.

Animals↗
PubMed1998

Testicular hormones do not regulate sexually dimorphic Pavlovian fear conditioning or perforant-path long-term potentiation in adult male rats.

We recently reported that Pavlovian fear conditioning and hippocampal perforant-path long-term potentiation (LTP) are sexually dimorphic in rats. Males show greater contextual fear conditioning, which depends on the hippocampus, as well as greater hippocampal LTP. In order to examine the role of circulating gonadal hormones in adult male rats, animals were castrated in two experiments, and Pavlovian fear conditioning and in vivo perforant-path LTP were examined. It was found that sexually-dimorphic LTP and fear conditioning are not regulated by the activational effects of testicular hormones in adult male rats. That is, in every respect, castrated male rats were similar to intact male rats in Pavlovian fear conditioning and hippocampal LTP. It is likely that sexual dimorphism in this system is established earlier in development by the organizational effects of gonadal hormones.

Animals↗
PubMed1998

Sexual differentiation of avian brain and behavior: current views on gonadal hormone-dependent and independent mechanisms.

Gonadal hormones are known to act during development to establish permanent sex differences in the anatomy and function of the vertebrate brain. They also act on the adult brain to activate reproductive behaviors. However, there are wide gaps in our understanding of how sexually dimorphic neural circuits translate into sex differences in behavior and other CNS functions. Moreover, not all sexually dimorphic properties of the adult brain can be attributed to known effects of gonadal hormones during development or adulthood, and factors other than gonadal steroids may contribute to these sex differences. This paper reviews sexual differentiation and the role of gonadal steroids and non-gonadal factors on sexually dimorphic development of the avian brain.

Animals↗
PubMed1997

A putative 5 alpha-reductase inhibitor demasculinizes portions of the zebra finch song system.

One model of the sexual differentiation of the zebra finch song system holds that both major metabolites of testosterone, dihydrotestosterone (DHT) and estradiol (E2), act together to masculinize the song system. To test this model, we administered a putative inhibitor of 5 alpha-reductase (MK-434) to decrease the synthesis of DHT from testosterone (T) in hatchling zebra finches. We tested MK-434's inhibition of 5 alpha-reductase, 5 beta-reductase, and aromatase in vivo and in vitro. In vivo, MK-434 significantly inhibited 5 alpha-reductase activity but also reduced the activities of 5 beta-reductase and aromatase. In vitro, MK-434 was extremely effective in inhibiting 5 alpha-reductase in the rat prostate but only slightly inhibited 5 alpha-reductase in the zebra finch telencephalon, where it also reduced aromatase and 5 beta-reductase activities. These results suggest that MK-434 might differentially influence the availability of androgenic and estrogenic substrates, depending on the relative abundance of these enzymes in brain. MK-434 demasculinized (decreased) the number and decreased the density of RA neurons but did not significantly affect any other sexually dimorphic aspect of the song system, including the volumes of RA, HVC, and Area X; the size of neural somata in IMAN, HVC, and RA; and the number of neurons in HVC and IMAN. The differential influence of MK-434 on sexually dimorphic characteristics suggests that the various sexually dimorphic characteristics of the song system (1) are sensitive to different hormones, depending on the characteristic; or (2) have different sensitivities to hormone levels, some being easily affected by slightly reduced hormone levels whereas others are not; or (3) have markedly different critical periods depending on the characteristic. Regardless of the reason(s) for differential effects on the sexually dimorphic characteristics of the song system, the data clearly suggest that steroid hormones play a role in the normal masculine development of the song system.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1997

Estrogen synthesis and secretion in the brown-headed cowbird (Molothrus ater).

Estrogens exert profound effects on vertebrate physiology and behavior. In most vertebrates, including birds, estrogens derived from ovarian tissue circulate at high levels during discrete periods of reproductive activity, and estrogen levels in males are low. In some songbirds (Passeriformes) plasma estrogens are high in both males and females. In the zebra finch, aromatase (estrogen-synthetase) is expressed abundantly in several regions of the male and female telencephalon and contributes to peripheral estrogen titers. To determine if this pattern of neural aromatase and estrogen synthesis is found in other songbirds, we have examined the patterns of estrogen synthesis in various tissues of another songbird, the common North American cowbird (Molothrus ater). Radiolabeled aromatizable androgenic substrate was injected in vivo or provided in vitro to telencephalic and gonadal tissue from adult male and female cowbirds. Estrogenic products were assayed in blood from the carotid artery and jugular vein, and in the telencephalon, ovary, and testes. Additionally, the presence of aromatase mRNA was studied in the brain using in situ hybridization. Radiolabeled androgenic substrate, injected in vivo, was readily converted to estrogens with higher amounts in the jugular compared to carotid blood, suggesting that the brain contains relatively high levels of aromatase. Further, radiolabeled androgens, provided in vitro to telencephalic, ovarian, and testicular tissue, resulted in the formation of radiolabeled estrogens. Aromatase mRNA is distributed widely in several areas of the cowbird telencephalon including the hippocampus, caudomedial neostriatum (including Field L), and nucleus taeniae. This pattern of neural aromatase expression resembles what we found previously in the zebra finch. Telencephalic aromatase may be characteristic of passerine songbirds and may function to provide local estrogenic cues to estrogen-sensitive neural loci, and/or contribute to peripheral estrogen titers in male and female songbirds.

Animals↗
PubMed1997

The activity and expression of aromatase in songbirds.

Songbirds have emerged as extremely important animals for investigating sex steroid hormone effects on the central nervous system. The masculinizing effects of exogenous estrogen on the neural circuits controlling song in female zebra finches are well documented. There is evidence that estrogens are necessary for the full activation of singing behavior in several species. These kinds of studies have led us and others to investigate the mechanisms whereby estrogens are made available to the brains of songbirds during development and adulthood. In this article, I review results of some of these studies examining the estrogen synthetic enzyme aromatase and its expression and activity in brain and in other tissues of songbirds. I will discuss some results and thoughts we have about the interactions of aromatase with the two remaining androgen-metabolizing enzymes in the avian brain, 5alpha-reductase, the enzyme that converts T into the active androgen 5alpha-dihydrotestosterone (DHT); and 5beta-reductase, the enzyme that converts T into the inactive 5beta-DHT. Finally, I will consider some ideas raised by these studies concerning potential sources of the androgen substrate for brain aromatization as well as some possible new functions that aromatase might be playing in the songbird telencephalon.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1997

Sex steroids and their actions on the birdsong system.

It is probably not surprising to most of us that the endocrine system plays a significant role in controlling the singing behavior of birds. We are familiar with the song of birds as a conspicuous acoustic feature of our environment during the avian breeding season. We often witness song when it is produced by birds (males) that are aggressively establishing and defending territories and that are advertising to available females. Thus, it is easy to imagine that song is likely to be stimulated by gonadal hormones. However, the ways in which gonadal sex steroids influence the various parts of the brain at various stages of the bird's life to influence song are complex and far from being completely understood. In this review, I will highlight some of the significant discoveries that have contributed to our view that the songbird brain is a significant and dynamic target of sex steroids. I will also describe what we have learned about properties of the endocrine system and the brain and how they each contribute to making androgens or estrogens available to particular parts of the songbird brain. Finally, I will describe some new research directions that may help answer some unresolved issues about hormonal effects on the songbird brain.

Animals↗
PubMed1996

3 beta-hydroxysteroid dehydrogenase/isomerase and aromatase activity in primary cultures of developing zebra finch telencephalon: dehydroepiandrosterone as substrate for synthesis of androstenedione and estrogens.

3 beta-hydroxysteroid dehydrogenase/delta 5 - delta 4 isomerase (3 beta-HSD) activity was measured in primary dissociated cell cultures prepared from telencephalons of developing zebra finches. 3 beta-HSD activity was confirmed after cultures were incubated with [7-3H]pregnenolone (Preg) or (1,2,6,7-3H-) dehydroepiandrosterone (DHEA) and 3H-progesterone (Prog) and 3H-androstenedione (AE) were detected in the medium. Product identity was confirmed by recrystallizations and by HPLC analysis. When DHEA was used as substrate, 3H-estradiol and 3H-estrone were also detected in the culture medium, presumably derived from the aromatization of 3H-AE or 3H-T produced from 3H-DHEA. To test this idea, cultures were incubated with 3H-DHEA together with radioinert AE or with fadrozole HCl, a potent and specific aromatase inhibitor. In the presence of radioinert AE, 3H-AE increased but metabolites of 3H-AE decreased in the media; in the presence of fadrozole, 3H-estrogens decreased but 3H-AE and its androgenic metabolite 3H-5 beta-androstanedione increased. These data demonstrate 3 beta-HSD activity in the songbird brain. The presence of Prog and estradiol in these cultures suggest that Preg and DHEA can potentially serve as substrates for the ultimate formation of active sex steroids in the songbird telencephalon.

3-Hydroxysteroid Dehydrogenases↗
PubMed1996

Independent differentiation of sexual and social traits.

In the process of development in mammals and birds, an individual becomes either male or female. The dogma of sexual differentiation explains this development in terms of gonadal secretions, which direct the growth of an organism along masculine or feminine lines. However, this type of bimodality is not found in all traits that are displayed in reproductive contexts. For example, aggressive behaviors displayed during the breeding season can be expressed by both males and females. Furthermore, this reproductive aggression is highly variable between individuals, regardless of sex. The development of variable traits like reproductive aggression is therefore likely to be influenced by factors in addition to gonadal secretions. We propose that social traits associated with reproduction can differentiate along pathways that are distinct from the developmental pathways of primary sexual traits.

Animals↗
PubMed1995

An atlas of aromatase mRNA expression in the zebra finch brain.

Neural conversion of androgen to estrogen by aromatase is an important step in the development and expression of masculine behavior in mammals and birds. In contrast to the low telencephalic levels of aromatase in adult mammals and nonsongbirds, the zebra finch telencephalon possesses high aromatase activity. This study maps, by in situ hybridization, cells that express aromatase mRNA in the adult zebra finch telencephalon, diencephalon, midbrain, and pons. High aromatase mRNA expression was observed in the caudal neostriatum, limbic archistriatum, and hypothalamus. The hippocampus, parahippocampal area, and hyperstriatum accessorium contained cells expressing moderate amounts of aromatase message. Weakly labeled cells were found in the rostral neostriatum, lobus parolfactorius, and mesencephalic reticular formation. These findings are consistent with aromatase activity measurements of zebra finch tissue and document with anatomical precision both the widespread expression of aromatase mRNA in the brain and novel sites of brain aromatase. This study identifies the caudal neostriatum as a major site of telencephalic aromatase. A previous survey (Gahr et al., 1993: J. Comp. Neurol. 327:112-122) of several avian species found that the presence of estrogen receptors in parts of the caudal neostriatum is unique to songbirds, which are the only birds to possess the elaborated telencephalic song system. Together, these findings suggest that the heightened estrogen synthesis and estrogen sensitivity of the passerine caudal neostriatum may have some functional relation with the telencephalic circuits responsible for song.

Animals↗
PubMed1995

Aromatase and 5 beta-reductase activity in cultures of developing zebra finch brain: an investigation of sex and regional differences.

Estrogen treatment of hatchling female zebra finches causes the masculine development of singing behavior and of the telencephalic brain regions involved in the control of song. However, early estrogen treatment of males also blocks masculine development of copulatory behavior, presumably controlled by diencephalic regions. In an effort to determine whether the differences in estrogen action are related to sex and regional differences in androgen metabolism (estrogen synthesis or androgen inactivation), we measured aromatase and 5 beta-reductase activity in dissociated-cell cultures made separately from the telencephalon, diencephalon, and also cerebellum of hatchling zebra finches under a variety of conditions. Cultures from all three brain regions express high levels of aromatase and 5 beta-reductase activity. Comparisons between telencephalic and diencephalic cultures of the activity and kinetics of aromatase suggest that the telencephalic cultures convert androgen to estrogen more efficiently than diencephalic cultures, which might be important in the differential action of estrogen in the two brain regions. However, the activity of neither aromatase nor 5 beta-reductase was significantly different between the sexes in either telencephalic or diencephalic cultures. Thus, comparisons between the sexes do not support the idea that differences in posthatching aromatase or 5 beta-reductase activity account for the pattern of sexual differentiation of the song and copulatory systems.

Androgens↗
PubMed1995

5 beta-reductase and other androgen-metabolizing enzymes in primary cultures of developing zebra finch telencephalon.

Enzymes in the avian brain irreversibly catalyze the conversion of androgens into either active metabolites (aromatase and 5 alpha-reductase) or inactive metabolites (5 beta-reductase), 5 beta-reductase is thought to influence the formation of active metabolites by reducing the concentration of androgenic substrate available for these reactions. However, because these enzymes have different regional, cellular and subcellular distributions in brain, the traditional method to measure enzyme activity in brain homogenates may be inaccurate because of artificial mixing of enzymes. Recently, we have prepared primary cell cultures from the telencephalons of developing zebra finches. Cell cultures offer the advantage that enzyme activity can be measured in live cells in which the relative distribution of enzymes may more closely reflect that found in vivo. We have previously reported that aromatase is expressed at high levels in these cultures, and that it is active in both neurons and in glia. Here we report on the activities of 5 alpha- and 5 beta-reductase in these cell cultures. Along with aromatase, 5 beta-reductase was expressed at high levels in these mixed cell cultures, including cultures highly enriched in glia. This suggests that 5 beta-reductase is present in non-neuronal cells in brain, possibly co-localized with aromatase. However, despite the presence of 5 beta-reductase, aromatase was detected reliably in vitro even when the concentration of substrate was low. Thus, 5 beta-reductase does not prevent the synthesis of estrogen in the telencephalon of developing zebra finches. By contrast, 5 alpha-reductase activity was very low or absent in these cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Androgens↗
PubMed1994

Fadrozole: a potent and specific inhibitor of aromatase in the zebra finch brain.

Aromatase and 5 beta-reductase activity are expressed at high levels in the zebra finch brain, especially in the telencephalon. Aromatization of androgens to estrogens is thought to be a critical step in the organization and activation of avian sexual behaviors. In contrast, 5 beta-reductase is thought to be an inactivating enzyme, one that catalyzes the conversion of androgens to biologically inactive metabolites. To address the importance of aromatase activity in this system, it is necessary to find an effective and selective aromatase inhibitor, one that has little or no effect on other androgen-metabolizing enzymes. The potency and specificity of fadrozole hydrochloride as an aromatase inhibitor was tested in zebra finch telencephalon. The compound was tested in vitro in primary dissociated cell cultures made from hatching telencephalon and compared to a commonly used inhibitor, 1,4,6-androstatriene-3,17-dione (ATD). Untreated, these cultures express extremely high levels of aromatase and 5 beta-reductase activity and therefore allow sensitive measurement of the effectiveness of inhibitors. Aromatase activity was also measured in homogenates of adult telencephalon following in vivo fadrozole injection. Finally, aromatase and 5 beta-reductase activity were quantified in zebra finch telencephalon following similar intramuscular injections in 4- to 6-day-old birds. In all three cases, fadrozole was highly effective in reducing aromatase activity. Fadrozole increased 5 beta-reductase activity, presumably due to an increase in available substrate, but had no inhibitory effect on the enzyme. ATD was less effective in inhibiting aromatase, and it also inhibited 5 beta-reductase activity at high concentrations.

Androstatrienes↗
PubMed1994

Isolation and characterization of a zebra finch aromatase cDNA: in situ hybridization reveals high aromatase expression in brain.

The brain regions controlling song are much larger in male than in female zebra finches. This sex difference is thought to arise developmentally when the male's brain is exposed to higher levels of estrogen. The synthesis of estrogen from androgen is catalyzed by aromatase, a key enzyme implicated in song development in the zebra finch. To study the role of estrogen synthesis in the organization of brain regions responsible for song, a cDNA encoding aromatase was isolated from a zebra finch ovarian cDNA library. The 3188 bp cDNA contains a 1527 bp open reading frame with 5' and 3' untranslated regions of 116 bp and 1545 bp, respectively. The deduced polypeptide is 509 amino acids in length, and is highly homologous to aromatases reported for chicken (92%), human (72%), mouse (70%), rat (69%) and trout (53%). Northern blot analysis revealed 5.4 kb, 4.8 kb and 3.2 kb aromatase mRNAs in brain and ovary. In situ hybridization histochemistry revealed the expression of aromatase mRNA in ovarian thecal cells of some, but not all, follicles, suggesting that aromatase gene expression is regulated during follicular maturation. In the adult brain, the distribution of aromatase mRNA was surprisingly widespread, and included the preoptic area, hypothalamus, hippocampus and neostriatum. By contrast, little aromatase mRNA expression was noted in the song nuclei (HVC, RA, area X). This study, the first description of aromatase mRNA expression by in situ hybridization in the brain of any species, identifies a surprisingly large number of cells that express aromatase mRNA in the zebra finch telencephalon. This pattern may be a unique feature of all songbirds.

Amino Acid Sequence↗
PubMed1994

Neuronal and non-neuronal aromatase in primary cultures of developing zebra finch telencephalon.

Estrogenic metabolites of circulating androgens have important effects on the organization and activation of neural circuits controlling reproductive behavior and physiology in males of many vertebrate species. Previous studies indicate that aromatase, the enzyme that converts androgens to estrogens, is expressed most abundantly in neurons in limbic brain regions. Songbirds are unique in that aromatase is expressed at unusually high levels throughout the telencephalon of both males and females. We assume that estrogens formed in the telencephalon itself masculinize neural circuits controlling song, since the brain is a major source of circulating estrogens in adult males. However, the cellular localization of telencephalic aromatase in songbirds remains unknown. We have established primary cultures from telencephalons of developing zebra finches and found aromatase activity (conversion of 3H-androstenedione or 3H-testosterone to 3H-estrone plus 3H-estradiol) at some of the highest levels reported for brain tissue of any species. Both neurons and glia were identified in these cultures based on cell morphology and labeling by specific immunohistochemical markers. However, when culture conditions were manipulated to reduce the incidence of either neurons or glia by varying the age of cultures or their plating density, treating with the neurotoxin kainic acid, physically shaking off loosely attached neurons, or preparing cultures in media that encouraged enrichment of neurons, high levels of aromatase persisted. Furthermore, Northern blot analysis of total RNA extracted from enriched neuronal or glial cultures indicated the presence of aromatase mRNA in both cell preparations. In situ hybridization with a zebra finch aromatase cDNA probe conjugated to digoxigenin showed the cultures contained darkly labeled neurons and lightly labeled non-neuronal cells, presumably astrocytes. We conclude that aromatase is expressed in both neuronal and non-neuronal cells in these cultures, suggesting that both cell types may also express the enzyme in vivo. The presence of aromatase outside of neurons suggests that glia may be targets of estrogen action or that glia may supply some estrogen to the estrogen-sensitive neural circuits in this species.

Aging↗
PubMed1993

Sexual differentiation of brain and behavior: the zebra finch is not just a flying rat.

In rats and other mammals, sex differences in the brain and behavior result from differential secretions of gonadal steroid hormones during early critical periods of neural development. We review the experimental results that support current ideas about the mechanisms of sexual differentiation in mammals, and then apply the same experimental analysis to the study of sexual differentiation of the neural song circuit in the zebra finch (Poephila guttata), a passerine song bird. Administration of estrogen to young female zebra finches causes the female to develop a more masculine song system and to sing as an adult. This estrogenic masculinization is similar to that found for copulatory behavior in mammals. However, striking differences emerge in other aspects of the sexual differentiation process. Experiments that use endocrine agents to block the masculine development in genetic males have so far failed. Moreover, the brain of zebra finches has an unusually high expression of aromatase (estrogen synthetase) in the telencephalon, and estrogen synthesized in the brain from androgen is released into the general circulation. These results suggest that the brain is the primary source of estrogen in the body. If so, then a further understanding of sexual differentiation requires more information on the factors that regulate the cerebral synthesis of estrogen.

Animals↗
PubMed1993

Estrogen synthesis in vivo in the adult zebra finch: additional evidence that circulating estrogens can originate in brain.

Aromatase activity is abundant in limbic and nonlimbic neural structures in zebra finches, especially in the telencephalon near neural circuits that control singing behavior. Also, male songbirds can have high estrogen levels in blood. Because it is difficult to detect aromatase activity in other male tissues, we have postulated that the brain itself is the source of the estrogen present in the blood of males. Previously, we developed methods to measure estrogen synthesis in vivo by injecting [3H]androgen into the systemic circulation or directly into tissues and then determining the quantity of [3H]estrogen entering or leaving the brain. Our results support our hypothesis that the brain is the primary site of estrogen synthesis in males of this species. Here, we confirm that [3H]estrogen emanates from the brain after systemic [3H]androgen injection by showing that the presumptive estrogen in jugular plasma is significantly reduced by treatment with an aromatase inhibitor. In females, but not males, estrogens can be found in large amounts in carotid plasma after systemic androgen injection, presumably due to ovarian estrogen synthesis. However, carotid estrogen levels are variable in females, so it is difficult to determine if the female brain contributes estrogen to blood as is seen in males. The data presented here suggest that the adrenal of males is not an important site of aromatization, because [3H]estrogens are undetected in the adrenals of castrated or sham-castrated males after adrenal injections of [3H] androgen. Nevertheless, at least 2.8- to 17.3-fold more [3H]estrogen was present in the jugular than in the carotid of castrated males after systemic injection of [3H]androgen. Because androgen levels were in the physiological range, we conclude that the brain routinely enriches the estrogen content of blood in normal adult males of this species.

Adrenal Glands↗