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Biomedical subjects

R W Lea

Publications and source records attributed to R W Lea.

At least 19 recordsLinked to original sources
PubMed2005

Aromatase inhibition abolishes courtship behaviours in the ring dove (Streptopelia risoria) and reduces androgen and progesterone receptors in the hypothalamus and anterior pituitary gland.

The aim of this study was to determine in the ring dove, the effects of aromatase inhibition on the expression of aggressive courtship and nest-soliciting behaviours in relation to the distribution of cells containing immunoreactive androgen (AR) and progesterone (PR) receptor in the hypothalamus and pituitary gland. Isolated sexually experienced ring doves were transferred in opposite sex pairs to individual breeding cages, and then injected with the aromatase inhibitor, fadrozole (four males and four females), or saline vehicle (four males and four females) for 3 days at 12 hourly intervals. Saline-injected control males displayed aggressive courtship behaviours (bow-cooing and hop-charging) and nest-soliciting throughout the study, and control females displayed nest-soliciting. By day 3, fadrozole treatment resulted in the disappearance of all these behaviours and in a decrease or disappearance of AR and PR in the anterior pituitary gland, and in the nucleus preopticus paraventricularis magnocellularis (PPM), nucleus preopticus medialis (POM), nucleus hypothalami lateralis posterioris (PLH), and ventral, lateral and dorsal nucleus tuberalis in the hypothalamus (VTu, LTu, DTu). In the nucleus preopticus anterior (POA), fadrozole treatment decreased AR in both sexes and decreased PR in females but not in males. Cells containing co-localized nuclear AR and PR were found in all hypothalamic areas examined, and in the anterior pituitary gland. Fadrozole is suggested to reduce the local availability of estrogen required indirectly for the induction of AR, and except in cells containing PR in the male POA, for the direct induction of PR. It is suggested that aggressive courtship behaviour is terminated by "cross talk" between aromatase-independent PR and aromatase-dependent AR co-localized in neurons in the POA. Aromatase-independent PR may increase in the male POA in response to visual cues provided by a partner. Aromatase-dependent PR in the POM, and basal hypothalamus may play a role in the facilitatory effect of progesterone on estrogen-induced nest-orientated behaviours.

Animal Communication↗
PubMed2001

Changes in central steroid receptor expression, steroid synthesis, and dopaminergic activity related to the reproductive cycle of the ring dove.

This review examines possible neural mechanisms involved in the expression of parental behavior in the ring dove, Streptopelia risoria. This avian species has proved an excellent animal model for studies concerning endocrine-behavior interactions for many years. Studies were performed to localize the expression of central androgen and progesterone receptor in both sexes. Expression of androgen receptor (androgen receptor immunoreactivity, AR-ir) was widespread but increased, similarly in both sexes, with increasing day-length. Progesterone receptor-immunoreactivity (PR-ir) was more localized in several discrete areas of the hypothalamus. Similarly, no sex differences were observed in PR-ir, and expression increased in birds maintained on long days. AR-ir demonstrated dramatic changes over the breeding cycle, being greatest in courting birds and almost undetectable in parenting birds of both sexes brooding their young. PR-ir showed a differential expression over the breeding cycle relative to its hypothalamic localization. PR-ir decreased in the tuberal hypothalamic area in brooding birds of both sexes; whereas in the preoptic area, PR-ir was maintained. Significant increases in dopaminergic activity during the parenting phase of the breeding cycle occurred in specific neural regions including the PVM and DMA. Studies demonstrated the ability of the diencephalon of both sexes of the ring dove brain to synthesize progesterone, with indications that in the male brooding dove, synthesis is increased. Finally, a model is presented that proposes a mechanism whereby these central systems may interact to result in the expression of full parental behavior in both sexes of the ring dove.

Animals↗
PubMed2001

Developmental changes in progesterone biosynthesis and metabolism in the quail brain.

We have recently demonstrated that the quail brain possesses the cholesterol side-chain cleavage enzyme (cytochrome P450scc) and 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4-isomerase (3beta-HSD) and produces pregnenolone, pregnenolone sulfate and progesterone from cholesterol. To elucidate the developmental changes in progesterone biosynthesis and its metabolism in the quail brain, we examined the expression and activity of 3beta-HSD and progesterone metabolite(s) during embryonic and post-hatched ages. Both the progesterone concentration and 3beta-HSD mRNA expression in the brain were almost constant during embryonic and post-hatched ages. The conversion of pregnenolone to progesterone (net 3beta-HSD enzymatic activity) was also constant during development and at maturity. However, without radioinert progesterone, the production of progesterone was drastically reduced in the embryonic brain, indicating active progesterone metabolism at the embryonic stage. Biochemical analysis together with HPLC and TLC revealed that only the embryonic brain actively produced 5beta-dihydroprogesterone from progesterone. Thus, progesterone production may be constant during embryonic and post-hatched development and in adulthood, whereas 5beta-dihydroprogesterone may be produced actively only in embryonic life due to 5beta-reductase.

Aging↗
PubMed2001

Androgen receptor immunolocalization in brains of courting and brooding male and female ring doves (Streptopelia risoria).

Nuclear androgen receptors (ARs) were localized immunocytochemically in the brains of courting and brooding male and female ring doves (Streptopelia risoria). AR immunoreactivity (AR-ir) in courting birds was localized in cell nuclei in the telencephalon, diencephalon, and mesencephalon. In the anterior hypothalamus, high density of AR-ir was concentrated in several nuclei including the nucleus lateralis hypothalami, nucleus periventricularis magnocellularis, nucleus preopticus anterior, nucleus preopticus medialis, and nucleus preopticus paraventricularis magnocellularis. In the posterior hypothalamus, areas showing high density of AR-ir included the nucleus lateralis hypothalami posterioris, nucleus medialis hypothalami posterior, nucleus ectomamillaris, nucleus mamillaris lateralis, and nucleus tuberis. No sex differences in the density or localization of AR-ir were observed. Compared to brains from courting birds, AR-ir density was either extremely low or absent in most brain regions of brooding birds. It is concluded that in the dove, central ARs are closely associated with the sexual stages of the reproductive cycle.

Animals↗
PubMed1999

Expression and activity of 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4-isomerase in different regions of the avian brain.

Recently, we have demonstrated, using biochemical and immunochemical methods, that the quail brain possesses the cholesterol side-chain cleavage enzyme (cytochrome P450scc) and produces pregnenolone and its sulfate ester. To clarify progesterone biosynthesis in the avian brain, therefore, we examined the expression of messenger RNA (mRNA) encoding for the enzyme 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4-isomerase (3beta-HSD) and its enzymatic activity using the quail. RT-PCR analysis together with Southern hybridization indicated the expression of 3beta-HSD mRNA in the brain of sexually mature birds but with no clear-cut sex difference. Employing biochemical techniques combined with HPLC analysis, the conversion of pregnenolone to progesterone was found in brain slices of mature males. Progesterone biosynthesis was increased in a time dependent manner and completely abolished by trilostane, a specific inhibitor of 3beta-HSD. The enzymatic activity of 3beta-HSD was greatest in the cerebrum and lowest in the mesencephalon. A specific RIA indicated that progesterone concentrations in the different brain regions closely followed the level of 3beta-HSD activity. High levels of progesterone concentration were observed in the diencephalon and cerebrum with lowest values in the mesencephalon. Progesterone levels in the brain regions were significantly higher than those in the plasma. These results suggest that the avian brain possesses not only cytochrome P450scc but also 3beta-HSD and produces progesterone. It is also indicated that progesterone biosynthesis in the avian brain may be region-dependent.

3-Hydroxysteroid Dehydrogenases↗
PubMed1999

An analysis of physiological mechanisms underlying the antigonadotropic action of intracranial prolactin in ring doves.

Intracerebroventricular (ICV) injections of prolactin (PRL) exert potent antigonadal and antigonadotropic effects in ring doves (Streptopelia risoria) at doses that are insufficient to stimulate prolactin-dependent crop growth. To explore the physiological basis of these effects, we tested the ability of ICV-injected PRL to influence pituitary responsiveness to chicken gonadotropin-releasing hormone-I (cGnRH-I) and to alter GnRH content and concentration in the preoptic area (POA) and median eminence (ME). cGnRH-I-induced changes in plasma LH were monitored by radioimmunoassay (RIA) in photostimulated male doves after they received five daily ICV injections of ovine PRL (1 microg/2 microl) or saline vehicle. Although PRL treatment reduced basal plasma LH levels and testes weight, it did not reduce the amount or alter the pattern of LH released in response to a bolus injection of cGnRH-I. This suggests that ICV PRL does not suppress LH by reducing pituitary responsiveness to GnRH. In two subsequent studies, GnRH content (ng/region) and concentration (pg/microg protein) in the POA and ME were measured in male doves by RIA and by competitive enzyme immunoassay after 5 days of ICV PRL or vehicle treatment. Although ICV PRL reduced plasma LH levels in both studies, no significant PRL-induced alterations in GnRH content or concentration were apparent. In a final study, PRL-treated female doves had lower plasma LH levels than vehicle-treated control females at 12 and 24 h after a single ICV injection. GnRH content of the POA was also lower in PRL-treated females than in controls at 24 h. However, the two treatment groups did not differ in POA or ME GnRH content at earlier postinjection sampling intervals. Analysis of GnRH concentration data revealed no treatment group differences in either region at any sampling interval (1, 6, 12, or 24 h post-PRL injection). Collectively, these results are consistent with the idea that ICV-injected PRL acts at the level of the CNS to inhibit the reproductive system. However, the nature of the alterations involved remains to be clarified. Plausible hypotheses are (1) that ICV PRL suppresses the gonadal axis by influencing the activity of GnRH neurons at brain sites other than the POA or ME or (2) that PRL alters the synthesis, storage, degradation, and/or release of GnRH in the POA or ME, but the dynamic changes involved are not reflected in integrated, steady-state measures such as peptide content or concentration in tissue.

Animals↗
PubMed1999

Neurosteroid biosynthesis in vertebrate brains.

In mammals, neurosteroids are now known to be synthesized de novo in the brain as well as other areas of the nervous system through mechanisms at least partly independent of the peripheral steroidogenic glands. However, limited information is available on neurosteroids in non-mammalian vertebrates. We therefore have attempted to demonstrate neurosteroid biosynthesis in the brain of birds and amphibians. These vertebrate brains possessed the steroidogenic enzymes, cytochrome P450 side-chain cleavage enzyme (P450scc) and 3beta-hydroxysteroid dehydrogenase/delta5-delta4-isomerase (3beta-HSD), and produced pregnenolone, pregnenolone sulfate ester and progesterone from cholesterol. Significant seasonal changes in neurosteroids in the brain were observed in seasonally breeding vertebrates. In addition, we attempted to identify the cell type involved in neurosteroidogenesis in mammalian and non-mammalian vertebrates in order to understand the physiological role of neurosteroids. Glial cells are generally accepted to be the primary site for neurosteroid formation, but the concept of neurosteroidogenesis in brain neurons has up to now been uncertain. We recently demonstrated neuronal neurosteroidogenesis in the brain and indicated that the Purkinje cell, a typical cerebellar neuron, actively synthesizes several neurosteroids de novo from cholesterol in both mammals and non-mammals. This paper summarizes the advances made in our understanding of neurosteroid biosynthesis, including neuronal neurosteroidogenesis, in a variety of vertebrate types.

Aging↗
PubMed1998

Control of luteinizing hormone and prolactin secretion in birds.

In birds, the secretion of luteinizing hormone (LH) and prolactin is controlled by the releasing hormones, gonadotrophin releasing hormone-I (GnRH-I) and vasoactive intestinal polypeptide (VIP), respectively. The secretion of LH is depressed and the secretion of prolactin is at its highest, during incubation/brooding young and during the development of reproductive photorefractoriness. In incubating domestic chickens, decreased LH and increased prolactin secretion are correlated with decreased hypothalamic GnRH-I mRNA and increased hypothalamic VIP mRNA. Increased plasma prolactin contributes to the suppression of LH secretion in incubating poultry, possibly acting at the levels of the hypothalamus and the anterior pituitary gland. During the development of photorefractoriness, decreased plasma LH and increased plasma prolactin are correlated with decreased hypothalamic GnRH-I and increased hypothalamic VIP. In birds exposed to seasonal changes in daylength, the seasonally maximal concentrations of plasma prolactin associated with the development of photorefractoriness can be explained, in part, by the saturation daylength for photoinduced prolactin release being reached in late spring/mid summer. During the development of photorefractoriness, high concentrations of plasma prolactin play a role in ensuring the rapid regression of the gonads but are not essential for the maintenance of the condition.

Animals↗
PubMed1998

Behaviour and hormone concentrations in nest deprived and renesting hens.

1. Nest and egg deprivation is a procedure traditionally used to disrupt incubation behaviour expression in commercial flocks. The aim of this study was to establish how nest deprivation affects the subsequent changes in behaviour and hormone secretion and whether readiness to renest is related to hormone concentration before and during nest deprivation. 2. Incubating broiler breeder hens were deprived of their nest either in their familiar environment, by blocking the nest entrance in the home cage or in an unfamiliar environment, by transfer in a different cage. After 3 days of nest deprivation, next access was allowed and readiness to incubate tested. 3. Both methods of nest deprivation resulted in the expression of similar behaviours associated with the disruption of incubation, and similar increases in plasma LH and oestradiol and decreases in plasma prolactin. The percentage (approximately 70%) of hens which renested after either method of nest deprivation did not differ significantly. 4. Readiness to renest was not related to the concentrations of plasma prolactin measured before or during the 3 days of nest deprivation. However, hens which would later renest could be identified by using behavioural criteria. They emitted more avoidance trills before, and sat on the wire floor for longer periods during, the nest deprivation period than the others.

Animals↗
PubMed1997

Localization of progesterone receptor in brain and pituitary of the ring dove: influence of breeding cycle and estrogen.

An immunocytochemical method was used in male and female ring doves (Streptopelia risoria) to localize progesterone receptor immunoreactivity (PR-ir) in the brain and anterior pituitary gland in nonbreeding, incubating, brooding, and estrogen (E2)-treated nonbreeding birds. Progesterone receptor was found in four regions of the brain in males and females: the preoptic area (POA), nucleus preopticus paraventricularis magnocellularis (PPM), nucleus hypothalami lateralis (PLH), and the tuberal region (TR). Quantitative analysis demonstrated that the density of cell nuclei containing PR-ir in the POA, PPM, and PLH in brooding birds was significantly higher than in E2-treated doves or in birds at other stages of the reproductive cycle. The density of PR-containing cell nuclei in the TR of male ring doves was significantly higher on day 1 of incubation than in nonincubating males. In brooding birds, there was a significant decrease in PR-ir in the TR, particularly in the ventral region where nuclei containing PR-ir disappeared. In the anterior pituitary gland the density of cell nuclei containing PR-ir was higher in females than in males at the onset of incubation. E2 treatment resulted in an increase in the density of PR-containing cell nuclei in both males and females. Brooding females had a lower concentration of PR-containing cell nuclei than did females at other stages of the breeding cycle. It is suggested that progesterone receptor in the POA mediates the expression of incubation behavior while progesterone receptor in the TR is involved in the control of neuroendocrine function. The source of estrogen which increases PR appears to be of central nervous origin in the male and may be predominantly peripheral in the female.

Animals↗
PubMed1997

Melatonin and the phagocytic process of heterophils from the ring dove (Streptopelia risoria).

A functional connection between the pineal gland and the immune system in mammals and birds has been established. This study investigates the effect of melatonin upon the non-specific immunity of heterophils isolated from the ring dove. The different stages of the phagocytic process: adherence to nylon fiber, spontaneous and induced mobility, ingestion of latex beads and digestion were evaluated for heterophils incubated in the presence of 5, 25, 50, 75, or 100 microM of melatonin. In addition, the chemoattractant power of the hormone for heterophils was studied. The 100 microM melatonin dose possessed a significant chemoattractant ability for heterophils whilst ingestion of latex particles was enhanced at all doses studied. The superoxide anion level, as measured by the free radicals produced during the metabolic burst, is decreased after incubation with 100 microM of melatonin.

Animals↗
PubMed1996

Secretion and serotonin release in the isolated rat lacrimal gland: the effects of substance P and calcitonin gene-related peptide.

A close anatomical relationship between nerves containing substance P and calcitonin gene-related peptide (CGRP) and mast cells containing serotonin has been demonstrated in the rat lacrimal gland. This study investigates the potential for peptidergic regulation of lacrimal mast cells by examining the actions of substance P, CGRP and serotonin on protein and peroxidase secretion from isolated lacrimal segments and on substance P and CGRP to release serotonin from the lacrimal mast cells. Substance P, CGRP and serotonin evoked marked increases in total protein and peroxidase from the lacrimal. Sodium cromoglycate, a mast cell stabilizer, significantly reduced or blocked the secretory responses elicited by these agonists. Chromatographic analysis using electrochemical detection revealed that substance P, but not CGRP, augmented the release of serotonin from the gland. The substance P evoked peroxidase secretion and serotonin release was blocked by CGRP and by sodium cromoglycate. These results support a role for mast cells in the regulation of lacrimal secretion and suggest a novel regulatory interaction between substance P and CGRP in the control of lacrimal function through a neuro-immune interaction.

Animals↗
PubMed1996

The effect of ovariectomy on concentrations of plasma prolactin and LH and parental behavior in the domestic fowl.

The role of ovarian hormones in the expression of parental behavior and in the regulation of LH secretion was investigated in incubating commercial meat-type hens. After ovariectomy, incubating hens continued to incubate eggs normally and brooded day-old chicks given to replace eggs, in a manner similar to sham-ovariectomized control hens. The concentration of plasma LH increased significantly in incubating hens after ovariectomy while the concentration of plasma prolactin remained high. Plasma LH remained depressed in sham-ovariectomized incubating control hens. The increase in plasma LH in incubating hens after ovariectomy (3.92 +/- 0.7 ng/ml) was less than that following the ovariectomy of nonincubating, nonlaying hens (5.3 +/- 1.2 ng/ml). The two groups of hens differed in that plasma prolactin concentrations were high (527 +/- 7.4 ng/ml) in the incubating hens and low (70 +/- 9 ng/ml) in the nonincubating hens. Nest deprivation resulted in an increase in plasma LH in both ovariectomized and sham-ovariectomized incubating hens with a significantly larger increase occurring in the ovariectomized hens (8.5 +/- 1.41 ng/ml compared to 2.48 +/- 0.65 ng/ml). Nest deprivation resulted in a similar rapid decrease in plasma prolactin in both ovariectomized and sham-ovariectomized hens. Replacement of eggs with day-old chicks in ovariectomized or sham-ovariectomized incubating hens resulted in a rapid decrease in plasma prolactin and after 6 days, in an increase in plasma LH in the ovariectomized but not sham-operated hens. It is concluded that once incubation behavior is established, ovarian hormones are not required for its maintenance or the readiness to brood day-old chicks. Ovarian hormones do, however, suppress LH release during incubation while the high concentration of plasma prolactin supplements this suppression.

Animals↗
PubMed1996

Expression of fos-like immunoreactivity in the hypothalamus of the ring dove (Streptopelia risoria) at the onset of incubation.

Fos immunocytochemistry was used in the ring dove (Streptopelia risoria) to identify areas in the forebrain and hypothalamus which might be involved in the expression of incubation behaviour and the associated changes in neuroendocrine function. In this species, both sexes participate in incubation which becomes fully established after laying the second egg in the two egg clutch. The eggs hatch after 14-15 days incubation. Observations were made in both sexes 16-18 h after laying the first egg, 3-5 h after laying the second egg, and after 12-14 days of incubation. Non-breeding birds were used as controls. Fos-labelled cells were observed in several forebrain areas in both sexes irrespective of reproductive status but their numbers did not change during incubation. In the hypothalamus, very few fos-labelled cells were seen in non-breeding birds but appeared in both sexes in the nucleus preopticus medialis after egg laying and after 12-14 days of incubation. Fos-labelled cells also appeared in the nucleus tuberis (=arcuate nucleus) in both sexes after laying the first or second egg and less abundantly, after 12-14 days incubation. Additionally, in males only, fos-labelled cells were seen after the laying of the first and second eggs in the rostral area of the nucleus tuberis close to the walls of the third ventricle. These observations suggest that the fos-labelled cells in the nucleus preopticus medialis may be involved in the expression of incubation behaviour while those in the nucleus tuberis may be involved in the regulation of gonadotrophin or prolactin secretion.

Animals↗
PubMed1996

Effect of age of donor on the responsiveness of dispersed and cultured chicken anterior pituitary cells to GnRH-I.

1. The aim of this study was to devise a method to prepare and culture anterior pituitary cells from juvenile and adult chickens in order to investigate mechanisms controlling gonadotrophin-releasing hormone-I (GnRH-I)-induced luteinising hormone (LH) release in vitro. 2. The optimum culture medium for maintaining gonadotroph responsiveness to GnRH-I was bicarbonate-buffered and phenol red-free Medium 199 supplemented with 10% foetal calf serum. 3. Cultured pituitary cells from juvenile chickens were more responsive to GnRH-I than cells from adult cockerels, while no LH was released in response to GnRH-I from pituitary cells from laying hens. 4. Cultured pituitary cells from adult chickens of both sexes released LH in response to 12-O-tetradecanoyl-13-phorbol acetate (TPA), an activator of an enzyme involved in intracellular signalling, protein kinase C. 5. It is concluded that freshly-dispersed and cultured gonadotrophs from adult chickens do not regain their responsiveness to GnRH-I as well as freshly-dispersed and cultured gonadotrophs from juvenile chickens. It appears that the stimulus-secretion coupling pathway between the GnRH-receptor and the activation of protein kinase C in gonadotrophs from adult chickens is more easily disrupted by dispersion and culture than in gonadotrophs from juvenile chickens.

Aging↗