bluuebunny/crossref_metadata_2025 huggingface dataset:
Nitric Oxide Is Formed in a Subpopulation of Rat Pineal Cells and Acts as an Intercellular Messenger
Authors: Spessert Rainer , Layes Elisabeth , Hill Gabriele , Vollrath Lutz
Published: April 2003

In the rat pineal, formation of the second messenger cyclic GMP (cGMP) is under adrenergic control. Two important sequential steps mediate adrenergic signal transduction by cGMP, receptor-stimulated nitric oxide (NO) formation by the enzyme NO synthase I (NOS I), and NO-induced cGMP formation by the cytosolic enzyme guanylyl cyclase. With regard to the first step in cGMP transduction (i.e. NO formation) we found, by means of NOS I immunostaining and NADPH-diaphorase staining, that the presence of NOS I was restricted to a subpopulation of pineal cells, generally surrounded by NOS I-negative cells. Considering the fact that NO is able to permeate the cell membrane, the question arises whether the second step in cGMP signalling (i.e. NO-induced cGMP formation) is restricted to the NO-forming cell itself or takes place in neighboring cells as well. It was found that oxyhemoglobin, which scavenges NO but does not enter cells, strongly decreased adrenergic cGMP response. This suggests that a considerable part of adrenergically induced NO stimulates cGMP formation in neighboring cells. Our findings indicate that in the rat pineal the components of adrenergic cGMP transduction are largely distributed among two types of cells: NOS I-containing pineal cells where adrenoceptor stimulation induces NO formation, and neighboring cells in which NO stimulates cGMP formation.
Cellular Composition of the Adult Rat Anterior Pituitary Is Influenced by the Neonatal Sex Steroid Environment
Authors: González-Parra Silvia , Argente Jesús , García-Segura Luis Miguel , Chowen Julie A.
Published: April 2003

Growth hormone (GH) and prolactin (PRL) secretion differ significantly between adult males and females and this is due, at least in part, to the postpubertal hormone environment which affects GH and PRL gene expression, as well as somatotrope and lactotrope proliferation. However, the role of the neonatal steroid environment in this phenomenon is less well understood. We have used in situ hybridization to determine the number of GH and PRL mRNA containing cells, as well as the level of expression of these two hormones and of the pituitary transcription factor 1 (Pit-1). Neonatally castrated male rats that had been exposed to testosterone during the neonatal period, adulthood or during both periods, males castrated as adults, normal adult males and normal proestrous females were used. Orchidectomy of adult rats had no effect on the number of somatotropes or lactotropes, but significantly reduced GH and PRL mRNA levels. Neonatal castration significantly reduced the percentage of somatotropes and increased that of lactotropes in the adult male. In addition, GH and Pit-1 mRNA levels were reduced significantly, but PRL mRNA levels were not modified. Treatment of neonatally castrated males with testosterone during the neonatal period significantly increased the percentage of somatotropes and decreased the percentage of lactotropes compared to vehicle-treated animals. It also increased GH and Pit-1 mRNA levels, but did not affect PRL mRNA levels. Adult testosterone treatment significantly increased the percentage of both somatotropes and lactotropes, as well as GH, PRL and Pit-1 mRNA levels. Treatment of neonatally castrated males with testosterone during both the neonatal and adult periods returned the percentage of somatotropes and lactotropes, as well as GH, PRL and Pit-1 mRNA levels, to that of the intact male. These results suggest that, although the postpubertal steroid environment is important in determining anterior pituitary hormone synthesis and cellular composition, the neonatal steroid environment also plays an important role in this phenomenon.
Interaction between Leptin and Neuropeptide Y on in vivo Growth Hormone Secretion
Authors: Carro Eva , Seoane Luisa M. , Señaris Rosa , Considine Robert V. , Casanueva Felipe F. , Dieguez Carlos
Published: April 2003

Leptin, the product of the ob gene, is a recently discovered hormone secreted by adipocytes that regulates food intake and energy expenditure. Leptin has recently been shown to play a stimulatory role on GH secretion. The aim of the present study was to investigate whether leptin regulation of GH secretion was mediated by hypothalamic neuropeptide Y (NPY). We assessed the effect of leptin administration (10 µg, i.c.v.) and/or NPY (4 µg, i.c.v.) on fasted rats. Furthermore we administered leptin antiserum (10 µl, i.c.v.), anti-NPY serum (5 µl, i.c.v.) or normal rabbit serum (10 µl, i.c.v.) to freely moving fed rats. Spontaneous GH secretion was assessed over 6 h with blood samples taken every 15 min. Fed rats treated with anti-NPY serum exhibited a normal ultradian GH rhythm. However, administration of anti-NPY serum (5 µl, i.c.v., at 120 min) completely reversed the suppression induced by antileptin serum (10 µl, i.c.v., at 0 min) on plasma GH levels (area under the curve, AUC, 168 ± 72 vs. 1,287 ± 430 ng/ml/6 h; p < 0.01). In fasted rats, following NPY administration, GH levels remained suppressed throughout the 6 h studied. Besides, NPY administration completely blunted leptin-induced GH secretion as assessed by the AUC (28.5 ± 11 vs. 520 ± 220 ng/ml/6 h; p < 0.01). Thus, it is possible that NPY mediates the effects of leptin on GH secretion. Alternatively, leptin and NPY could act through parallel pathways to alter GH release with NPY overcoming the stimulatory effect exerted by leptin on plasma GH levels.
Allopregnanolone-Induced Modification of Presynaptic Basal and K+-Induced [3H]-Norepinephrine Efflux from Rat Cortical Slices during the Estrous Cycle
Authors: Belmar Jorge , Cuellar Claudia , Llona Isabel , Arancibia Sandor , Kusch Claudia , Tapia-Arancibia Lucía , Pinter Albin , Pérez Hernán
Published: April 2003

Superfused frontal slices of cerebral cortex were preloaded with [3H]-norepinephrine ([3H]NE). Basal [3H]NE efflux and K+-induced [3H]NE release were studied during the estrous cycle and in the presence of neurosteroids. Basal [3H]NE efflux showed estrous cycle-related variations, with lowest values found during estrus and diestrus II. Allopregnanolone (10–9 M) potentiated basal [3H]NE efflux from the 1st minute of its application; the effect of the steroid was still present after 20 min. This effect was also dependent upon the estrous cycle, since basal [3H]NE efflux was mainly increased during estrus diestrus I, and to a lesser degree only during proestrus. During diestrus II and after ovariectomy, basal [3H]NE efflux was no longer affected by the neurosteroid. In the presence of yohimbine (10–6 M), the effect of allopregnanolone on basal efflux was potentiated only during the first 3 min but vanished thereafter. Allopregnanolone (10–9 M) potentiated the K+-induced [3H]NE release during estrus, but pregnenolone (10–9 M) was ineffective, suggesting specificity of the neurosteroid. Yohimbine (10–6 M) also potentiated K+-induced [3H]NE release. When applied simultaneously with allopregnanolone (10–9M), a potentiating effect on [3H]NE release was observed. The present results suggest that allopregnanolone is a neurosteroid able to modulate norepinephrine release in the cerebral cortex in an estrous cycle-dependent manner, and that the effect could involve noradrenergic alpha-2 receptors.
In vitro Secretion of Gonadotropin-Releasing Hormone (GnRH) and [Hydroxyproline9]GnRH from the Rat Hypothalamus Exhibits a Differential Sensitivity to Castration and Second Messengers
Authors: Rochdi Laïla , Theraulaz Laurence , Enjalbert Alain , Gautron Jean-Pierre
Published: April 2003

The decapeptide [hydroxyproline9]GnRH (HypGnRH) has been characterized as an endogenous posttranslational product of the gonadotropin-releasing hormone (GnRH) precursor in a wide range of mammalian brains. Despite consistent biological effects, its secretion by the hypothalamus remains hypothetical. We report here in vitro secretion of HypGnRH and GnRH by the hypothalamus from intact and castrated male rats and provide evidence that they are differentially regulated. Both peptides were identified by two anti-GnRH antibodies of different specificities after separation under two high-performance liquid chromatography conditions. Calcium dependency of HypGnRH release was demonstrated under stimulation with KCl in the absence or presence of Ca2+, as well as with Bay K 8644, veratridine, methoxyverapamil, or tetrodotoxin. Activation of signaling pathways involving adenylate cyclase and protein kinases A and C (PKC) induced HypGnRH release. Expression of data as percentage of release over tissue stores revealed a two- to threefold higher release of HypGnRH than of GnRH under the different modes of stimulation used, except under PKC activation which triggered a comparable recruitment of both peptides. Castration selectively affected PKC-coupled GnRH secretion which showed a twofold lesser release than in intact rats, while the HypGnRH release was unaffected. We conclude that HypGnRH and GnRH are not secreted from the hypothalamus according to the same mechanisms.
Regulation of Prolactin Secretion by Dopamine and Vasoactive Intestinal Peptide at the Level of the Pituitary in the Turkey
Authors: Youngren Orlan M. , Chaiseha Yupaporn , El Halawani Mohamed E.
Published: April 2003

This study investigated the capability of dopamine (DA) to prevent avian prolactin (PRL) secretion by antagonizing the PRL-releasing factor, vasoactive intestinal peptide (VIP), at the level of the pituitary. To test this hypothesis, combined intracranial and intrapituitary infusions of DA, DA agonists, and VIP, plus electrical stimulation of the medial preoptic area (ES/POM), were used to characterize the actions of DA on PRL secretion in anesthetized laying turkey hens. Infused into the third ventricle at the rate of 10 nmol/min, DA induced a 2.8-fold increase in circulating PRL levels (63.8 ± 15.1 to 181.3 ± 44.3 ng/ml, p < 0.05), similar to the 3.1-fold PRL increase induced by ES/POM (65 ± 10.3 to 199.1 ± 57.3 ng/ml, p < 0.05). Infused into the anterior pituitary at the rate of 15 ng/min, VIP induced a 2.2-fold increase in PRL (78.6 ± 22.9 to 173.6 ± 39.5 ng/ml, p < 0.05). When DA (10 nmol/min) was infused into the anterior pituitary it completely blocked both ES/POM- and VIP-induced PRL secretion. The D2 DA receptor agonist R- (–) -Propylnorapomorphine HCl inhibited VIP-induced PRL secretion at the level of the anterior pituitary, allowing only an insignificant rise in PRL (54.8 ± 14.3 to 73.9 ± 21.6 ng/ml, p > 0.05), while the D1 DA receptor agonist (±)-SKF-38393 HCl failed to prevent VIP-induced PRL release, allowing PRL to rise 2.5-fold (49.1 ± 10.8 to 121.0 ± 34.8 ng/ml, p < 0.05). Pituitary infusion of DA, DA agonists or vehicle alone caused no change in PRL levels. The data showed that DA prevented avian PRL secretion by blocking the action of VIP at the level of the anterior pituitary. DA effected this blockade of PRL via D2 DA receptors residing within the anterior pituitary. The data also suggested that there were no stimulatory D1 DA receptors related to PRL secretion in the avian anterior pituitary.
Direct Action of Serotonin on Prolactin, Growth Hormone, Corticotropin and Luteinizing Hormone Release in Cocultures of Anterior and Posterior Pituitary Lobes: Autocrine and/or Paracrine Action of Vasoactive Intestinal Peptide
Authors: Balsa José Antonio , Sánchez-Franco Franco , Pazos Fernando , Lara José Ignacio , Lorenzo María Jesús , Maldonado Gonzalo , Cacicedo Lucinda
Published: April 2003

There is extensive evidence that serotonin (5-HT) is implicated in the neuroendocrine control regulating the secretion of several anterior pituitary hormones. It has also been reported that the posterior pituitary is necessary for prolactin (PRL) response to 5-HT as well as to suckling, in which 5-HT implication has been demonstrated. As we have previously shown that vasoactive intestinal peptide (VIP) mediates through an autocrine or paracrine action the PRL release induced by insulin-like growth factor I, thyrotropin-releasing hormone (TRH) and dopamine withdrawal, the aim of the present work was to determine whether 5-HT has a direct action on pituitary secretion and to study the possible role of pituitary VIP in this situation. Cells from the anterior pituitary lobe (AP) were cultured either alone or together with cells from the posterior pituitary lobe (PP). As melanotropes from PP express glucocorticoid receptors in vitro, both AP cultures and cocultures of AP/PP cells were incubated in the presence or absence of corticosterone (0.1 µg/ml), thus designing four experimental conditions. Then both AP and mixed cultures were incubated with 5-HT (100 nM) for 20, 45 and 180. The release of PRL, growth hormone (GH), corticotropin (ACTH) and luteinizing hormone (LH) was stimulated by 5-HT, but only in cocultures of AP/PP cells preincubated with corticosterone, whereas follicle-stimulating hormone and thyroid-stimulating hormone release was not modified. As AP cultures did not show any response to 5-HT, both in the presence or absence of corticosterone, and as melanotropes are the main cellular type present in the PP cultures, we studied the response of α-melanocyte-stimulating hormone (αMSH) to 5-HT in PP cells cultured with or without corticosterone. Serotonin did not modify αMSH release either in the absence or the presence of corticosterone. VIP release was also stimulated by 5-HT in the cocultures, and the time response profile was only similar to that of PRL. In order to study whether pituitary VIP is implicated in 5-HT action, cocultures preincubated with corticosterone were incubated in the presence of 5-HT, a VIP-receptor antagonist (VIP-At) or simultaneously with 5-HT plus VIP-At. PRL response to 5-HT was abolished by the simultaneous presence of VIP-At, whereas GH, ACTH and LH response remained unchanged. These data demostrate that: (1) 5-HT stimulates the secretion of PRL, GH, ACTH, LH and VIP acting directly at pituitary level on PP, probably by releasing an unidentified mediator from melanotropes; (2) glucocorticoids make the response of AP cells to 5-HT possible due to the presence of PP cells in the coculture; (3) PRL response to 5-HT is mediated through an autocrine and/or paracrine action of VIP.
Neuroendocrine-Related Effects of Long-Term, ‘Binge’ Cocaine Administration: Diminished Individual Differences in Stress-Induced Corticosterone Response
Authors: Sarnyai Zoltán , Dhabhar Firdaus S. , McEwen Bruce S. , Kreek Mary Jeanne
Published: April 2003

Acute cocaine administration activates behavioral and neuroendocrine processes associated with the stress response. However, much less is known about the effects of chronic, long-term cocaine administration on neuroendocrine adaptations and individual vulnerability to stress. We hypothesized that chronic ‘binge’ cocaine administration may serve as a chronic pharmacological stressor leading to a hyperactivity of the stress-responsive hypothalamic-pituitary-adrenal (HPA) axis and alterations in its feedback mechanisms. In order to test this hypothesis, the effects of long-term (3 and 6 weeks) ‘binge’ pattern cocaine administration (3×15 mg/kg cocaine, i.p., daily, during the early phase of the light cycle) on body weight, adrenal gland weight, basal and stress-induced activity of the corticosterone (CORT) and basal plasma testosterone (T) levels were measured. Both 3 and 6 weeks ‘binge’ cocaine administration decreased body weight gain, increased the weight of adrenal glands and increased basal CORT levels. Plasma T levels were suppressed by both 3 and 6 weeks of cocaine treatment. No correlation was found between elevated CORT and low T levels at any time point. Neither chronic saline nor cocaine administration altered stress-induced CORT secretion. CORT levels 60 min following the restraint stress (recovery) were significantly lower than pre-stress basal levels after 3 and 6 weeks of cocaine, but not saline, administration. Moreover, initial individual differences in stress-induced CORT response, i.e. low and high responsivity to restraint prior to any saline or cocaine injections, were maintained in control rats but became diminished in cocaine-treated rats. These results indicate that chronic binge cocaine administration leads to sustained activation of the HPA axis and alters processes underlying individual vulnerability to stress.
The Role of N-Methyl-D-Aspartate Receptors in the Release of Adrenocorticotropin by Dynorphin A1–13
Authors: Szeto Hazel H. , Soong Yi , Wu Dunli
Published: April 2003

We previously reported that dynorphin A1–13 evokes a significant increase in plasma adrenocorticotropin (ACTH) after intravenous administration in the ovine fetus. This response was not sensitive to naloxone and was regulated differently from the response to U50,488H, a selective ĸ-opioid agonist. NMDA appears to play a role in many of the nonopioid actions of dynorphin. We therefore hypothesized that dynorphin A1–13 may release ACTH via N-methyl-D-aspartate (NMDA) receptors. To test this hypothesis, we have compared the ACTH response to dynorphin A1–13 and NMDA in the chronically-instrumented ovine fetus. Our data show that both dynorphin A1–13 (0.5 mg/kg) and NMDA (4 mg/kg) induced a significant release of immunoreactive ACTH in the late-term ovine fetus. The ACTH response to NMDA was of a smaller magnitude, but of longer duration, when compared to dynorphin A1–13. The response to both dynorphin A1–13 and NMDA was significantly attenuated by pretreatment with the noncompetitive NMDA antagonist, MK-801, but was not affected by antagonists of corticotropin-releasing hormone and arginine vasopressin. Finally, the ACTH response to both dynorphin A1–13 and NMDA were inhibited by dexamethasone. The results of this study indicate a role for NMDA receptors in the action of dynorphin A1–13, and suggest that NMDA may act directly at the level of the pituitary to release ACTH without the involvement of hypothalamic secretagogues.
Projections of the Estrogen Receptor-Immunoreactive Ventrolateral Hypothalamus to Other EstrogenReceptor-Immunoreactive Sites in Female Guinea Pig Brain
Authors: Turcotte JoanneC. , Blaustein JeffreyD.
Published: April 2003

The ventrolateral hypothalamus in female guinea pigs includes an estrogen receptor dense region adjacent to the ventromedial hypothalamus. This region is reciprocally connected with other estrogen receptor-containing areas suggesting that steroid hormone receptor-containing cells may be directly linked. Phaseolus vulgaris leucoagglutinin, an anterograde tract tracer, was specifically placed in this region with the aim of labeling some projections from estrogen receptor-containing neurons. These projections were colocalized immunocytochemically with the distribution of estrogen receptor-containing cells. Dense ventrolateral hypothalamic innervation was observed in some regions also containing a high concentration of estrogen receptor-containing cells. These regions included the medial preoptic area, the bed nucleus of the stria terminalis, the ventrolateral hypothalamus anterior and posterior to the injection site, and the midbrain central gray. A low density of ventrolateral hypothalamic fibers and terminals was observed in two regions rich in estrogen receptors, the amygdala and the arcuate nucleus. In general, ventrolateral hypothalamic fibers and terminals were present in all regions where estrogen receptors were found except the medial thalamus and habenular region. Labeled terminal boutons and perineuronal baskets were found around estrogen receptor-containing cells in most regions which contained estrogen receptor-containing cells. These close appositions were suggestive of synaptic contacts, suggesting that the ventrolateral hypothalamus may influence steroid-dependent behaviors via the modulation of estrogen receptor-containing cells. Furthermore, ventrolateral hypothalamic projections may include direct connections with estrogen receptor-containing cells, suggesting the presence of a network of interconnected estradiol-sensitive neurons involved in the regulation of estradiol-dependent functions.
Acute Intrahippocampal Injection of Human Interleukin-1β Stimulates the Anterior Pituitary POMC Transcription and Increases Plasma Levels of ACTH and Corticosterone in the Male Rat
Authors: Mélik Parsadaniantz S. , Daugé V. , Roques B.P. , Kerdelhué B.
Published: April 2003

It has been well documented that interleukin-1β (IL-1β) is a major mediator for recruiting the hypothalamo-pituitary-adrenal (HPA) axis following infectious disease. The recent localization of IL-1β receptors in neurons of the hippocampus provides further support for the role of IL-1β as a neurotransmitter/neuromodulator in the central nervous system. In this study, we investigated whether an acute intrahippocampal injection of IL-1β is able to rapidly stimulate HPA activity. Seven days after bilateral implantation of a guide cannula into the hippocampus, human IL-1β (10 ng/0.5 µl/side) was injected to freely moving male rats. Following this, animals were sacrificed at times 20, 45 and 90 min postinjection and a kinetic analysis of hIL-1β action on plasma ACTH and corticosterone (CORT) concentrations and nuclear processing of the anterior pituitary (AP) proopiomelanocortin (POMC) was conducted. Intrahippocampal administration of hIL-1β significantly increased both plasma ACTH and CORT concentrations at 45 and 90 min postinjection. This increase in ACTH concentration paralleled a rise in AP POMC gene transcription. Moreover, the increase in AP POMC primary transcript was followed by an increase in AP POMC intermediate processing RNA. However, at these times, no significant hIL-1β effect on the level of AP nuclear POMC mRNA was observed. Almost identical results were obtained after intraperitoneal injection of hIL-1β. In conclusion, our data demonstrates that the hippocampal IL-1β/IL-1β receptor is directly and rapidly implicated in HPA activation, in the same manner as that observed after intraperitoneal administration of hIL-1β. These results show that IL-1 action in the hippocampus could be of immunoneuroendocrine significance for the HPA axis activation during inflammatory states.
Hyporesponsiveness of the Pituitary to CRH during Slow Wave Sleep Is Not Mimicked by Systemic GHRH
Authors: Antonijevic Irina A. , Murck Harald , Frieboes Ralf-Michael , Holsboer Florian , Steiger Axel
Published: April 2003

During slow wave sleep (SWS) pituitary responsiveness to CRH is reduced. Since GHRH is involved in the promotion of SWS in humans and rats, it was examined whether the blunted CRH-induced ACTH and cortisol release during SWS could be mimicked by systemic GHRH. Young healthy men (n = 7) participated in 4 sleep-endocrine protocols: (A) lights off at 23.00 h, intravenous injection of 50 µg CRH during the first SWS period; (B) lights off at 01.00 h, injection of 100 µg GHRH at 23.00 h, followed by 50 µg CRH at 23.30 h; (C) lights off at 01.00 h, injection of 50 µg CRH at 23.30 h, and (D) lights off at 23.00 h, saline treatment only (= baseline condition). The sleep EEG was recorded during the lights off period and blood samples, collected every 20 min between 22.00 and 07.00 h, were assayed for GH, cortisol and ACTH. There was no significant difference in the sleep-associated GH peak between protocols. Plasma ACTH was significantly higher following CRH administration during wakefulness compared with CRH administration during SWS (protocols B and C vs. A; area under the curve (AUC) 23.00–03.00 h: 9.6 ± 4.8 and 7.3 ± 2.0 vs. 6.1 ± 1.1 ng/ml × min; p < 0.05), while there was no significant difference in plasma ACTH concentration between the baseline condition and protocol A (CRH administration during SWS). Similarly, cortisol was significantly enhanced compared with baseline following CRH during wakefulness only. CRH induced an increase in EEG activity in the sigma frequency range, both when it was administered during wakefulness and SWS, while this effect was reduced by pre-treatment with GHRH. In summary, our data suggest that (1) the blunted CRH-induced release of ACTH and cortisol during SWS is not mimicked by systemic GHRH administration, and (2) CRH enhances sigma EEG activity possibly via modulation of afferent pathways from the median eminence to the thalamus and this effect is reduced by pre-treatment with GHRH.
The Role of the Retrochiasmatic Area in the Control of Pineal Metabolism
Authors: Cipolla-Neto José , Skorupa AnaLúcia , Ribeiro-Barbosa ÉrikaR. , Bartol Ione , Mota Sandra Regina , Afeche SolangeCastro , Delagrange Philippe , Guardiola-Lemaitre Béatrice , Canteras NewtonSabino
Published: April 2003

The aim of the present investigation was to study the effect of neurotoxic ibotenic acid lesion of the retrochiasmatic area on the daily profile of pineal N-acetylserotonin and melatonin synthesis and on the pineal metabolic reactivity to nocturnal short-term retinal photostimulation. Groups of rats were killed 6 h after lights off either in the dark of immediately after being photostimulated for 1 or 15 min. Additionally, groups of rats were sacrificed at six different time points throughout the 24-hour light-dark cycle. The results suggested the presence of two functionally distinct territories in the retrochiasmatic area. The basal retrochiasmatic area, an area situated immediately ventral to the third ventricle, behind the suprachiasmatic nuclei and in front of the arcuate nucleus, is implicated in the nocturnal inhibitory process induced by short-term retinal photostimulation. The lateral retrochiasmatic area, which is situated immediately lateral to the anterior periventricular nucleus, below the anterior hypothalamic nucleus and in front of the ventromedial hypothalamic nucleus, is importantly involved in the control of the peak amplitude of the daily production of N-acetylserotonin and melatonin by the pineal gland.
In vivo Characterization of 11β-Hydroxysteroid Dehydrogenase in Rat Hippocampus Using Glucocorticoid Neuroendangerment as an Endpoint
Authors: Ajilore Olusola A. , Sapolsky Robert M.
Published: April 2003

11β-Hydroxysteroid dehydrogenase (11β-HSD) is the enzyme responsible for the interconversion of corticosterone (CORT) to 11-dehydrocorticosterone (11-DHC). CORT is an adrenal hormone secreted during the stress response and it has widespread effects in many different target tissues. In addition, CORT can exacerbate damage caused by neurological insults, such as kainic acid-induced seizures. In addition to its protective role in the kidney, 11β-HSD is also thought to play a role in steroid regulation in the brain. However, it is not known whether the enzyme is acting in vivo as a reductase or a dehydrogenase. If the enzyme is working as a reductase, converting 11-DHC to CORT, it has the potential to exacerbate neurotoxicity due to other agents. On the other hand, 11β-HSD could be neuroprotective if the enzyme is acting as a dehydrogenase, deactivating CORT by converting it into 11-DHC. To characterize the enzyme in vivo, we have utilized glucocorticoid neuroendangerment in the hippocampus as an indirect assay of 11β-HSD function. We have shown that 11-DHC can exacerbate kainic acid toxicity in adrenalectomized (ADX) rats and this exacerbation is blocked by the 11β-HSD antagonist, carbenoxolone; these findings suggest that 11β-HSD is working as a reductase in ADX rats. The presumptive reductase activity found in ADX rats was derived from both hippocampal and peripheral forms of the enzyme. In the presence of physiological levels of glucocorticoids, reductase activity was decreased and no dehydrogenase activity was detected. The present study demonstrates that 11β-HSD reductase activity, both in vivo and in vitro, occurs only in the presence of low levels of circulating glucocorticoids.
Effect of Selective Blockade of Catecholaminergic Alpha and Beta Receptors on Histamine-Induced Release of Corticotropin and Prolactin
Authors: Willems Edwin , Knigge Ulrich , Jørgensen Henrik , Kjær Andreas , Warberg Jørgen
Published: April 2003

We investigated the role of adrenergic receptors in histamine (HA)-induced release of corticotropin (ACTH) and prolactin (PRL) in conscious male rats. Specific α- or β-receptor antagonists were administered intracerebroventricularly in doses of 1 mmol at time –20 min, and HA (270 nmol), the H1 receptor agonist 2-thiazolylethylamine (2-TEA; 2,180 nmol) or the H2 receptor agonist 4-methylHA (4-MeHA; 790 nmol) were administered intracerebroventricularly at –15 min. The animals were decapitated at 0 min, and plasma was analyzed for ACTH and PRL. Administration of HA and the histaminergic agonists stimulated ACTH secretion equally, while only HA and the H2 receptor agonist stimulated PRL secretion. Pretreatment with the adrenergic receptor antagonists had no effect on the ACTH response to the histaminergic compounds. In contrast, the PRL response to HA or 4-MeHA was inhibited or prevented by the α-receptor antagonists phenoxybenzamine and phentolamine, the α1-receptor antagonist prazocin, the β-receptor antagonist propranolol and the β1-receptor antagonist atenolol, whereas the α2-receptor antagonist yohimbine or the β2-receptor antagonist ICI-118-551 had no effect. The study indicates that histaminergic neurons interact with the catecholaminergic neuronal system in regulation of PRL secretion, and that this interaction is dependent upon activation of α1- and β1-receptors. In contrast, histaminergic neurons stimulate ACTH secretion independently of adrenergic receptor activation.
Role of Glucocorticoids in the Response of the Hypothalamo-Corticotrope, Immune and Adipose Systems to Repeated Endotoxin Administration
Authors: Chautard Thierry , Spinedi Eduardo , Voirol Marie-Jeanne , Pralong François P. , Gaillard Rolf C.
Published: April 2003

The present study was designed to determine whether the glucocorticoid inhibitory feedback mechanism plays a role in the well-known tolerance of the neuroendocrine-immune axis response to repeated endotoxemia. Adult male rats underwent adrenalectomy (ADX) and were implanted with a subcutaneous corticosterone (compound B, CB, 75 mg) pellet, or sham operated and implanted with a placebo pellet. On the morning of day 8 after surgery (experimental day, D1), all rats received an intravenous injection of lipopolysaccharide (LPS) (25 µg/kg body weight) which was repeated daily until D5. Blood was drawn via intravenous indwelling catheters before (sample time zero) as well as 1, 2, 3 and 4 h after LPS treatment on D1, 3 and 5 for measurements of corticotropin (ACTH), CB, tumor necrosis factor-α (TNF-α) and leptin. In sham animals, tolerance to repeated LPS administration was complete by D5 for the corticotrope axis and the immune response. In addition, LPS was found to stimulate leptin secretion on day 1 in intact rats, an effect that also disappeared thereafter. ADX + CB rats showed only a partial tolerance of the corticotrope axis on D5, whereas tolerance of the immune response was similar to that found in sham animals. Interestingly, the acute stimulation of leptin secretion by LPS in ADX + CB rats was qualitatively similar to that of intact controls on D1, but plasma leptin levels were significantly reduced on D3 and 5 compared to controls. Our results demonstrate that the adrenal response tolerance of the hypothalamo-pituitary-adrenal axis to repeated endotoxemia. In addition, our finding that TNF-α secretion follows the same pattern in sham-operated and in adrenalectomized animals suggests that unlike the corticotrope axis, tolerance of the immune response does not depend upon stimulated CB levels. The decrease in circulating levels of leptin following ADX is consistent with the stimulatory effects of glucocorticoids on leptin secretion. However, our finding of an acute stimulation of leptin secretion by LPS in ADX + CB animals demonstrates that this effect of endotoxemia is at least partially glucocorticoid independent.
PC18, a Specific Aminopeptidase N Inhibitor, Induces Vasopressin Release by Increasing the Half-Life of Brain Angiotensin III
Authors: Réaux Annabelle , de Mota Nadia , Zini Sylvie , Cadel Sandrine , Fournié-Zaluski Marie-Claude , Roques Bernard P. , Corvol Pierre , Llorens-Cortès Catherine
Published: April 2003

Angiotensin III (AngIII), which is metabolized in vivo by aminopeptidase N (APN), was previously shown to be one of the main effector peptides of the brain renin-angiotensin system (RAS) in the control of vasopressin release. Recently, a potent APN inhibitor, PC18 (2-amino-4-methylsulfonyl butane thiol, methionine thiol), has been developed. In this study, we first checked the in vitro selectivity of PC18 towards APN, aminopeptidase A (APA) and aminopeptidase B (APB), three zinc metalloproteases with significant identity between their amino acid sequences. The Ki values of this compound on APN were found to be in the nanomolar range (Ki = 8.0 ± 1.7 nM) but it was 2,150 and 125 times less active on APA and APB, respectively. Secondly, we evaluated in vivo the effect of brain APN inhibition with PC18 on the inactivation of brain AngIII and on vasopressin secretion in mice. For this purpose, mice received [3H]AngII intracerebroventricularly in the presence or absence of the APN inhibitor PC18 (30 µg). At different times after the injection, [3H]AngIII levels were evaluated from hypothalamus homogenates after separation by cation-exchange chromatography. PC18 induced an accumulation of [3H]AngIII, increasing its half-life 3.9 times as compared with control values. In addition, the effect of PC18 on vasopressin release was studied in mice. PC18 (10–100 µg) was injected intracerebroventricularly, and plasma vasopressin levels were estimated by radioimmunoassay. PC18 increased vasopressin levels in a dose-dependent manner. The maximal increase in vasopressin release (+220%) is observed for a dose of PC18 of 100 µg and was inhibited 75% by the coadministration of the AngII receptor antagonist (Sar1–Ala8)-AngII (0.5 µg). These results indicate that in vivo, in the mouse brain, APN inhibition by PC18 increases the half-life of endogenous AngIII, resulting in an enhanced vasopressin release.
Cyclical Variations in the Abundance of Leptin Receptors, but not in Circulating Leptin, Correlate with NPY Expression during the Oestrous Cycle
Authors: Bennett Pamela A. , Lindell Kajsa , Wilson Carole , Carlsson Lena M.S. , Carlsson Björn , Robinson Iain C.A.F.
Published: April 2003

We have demonstrated previously that pharmacological doses of oestradiol decreased leptin receptor expression in the hypothalamus. We therefore analysed leptin receptor expression during the oestrous cycle in the rat, to establish if acute changes in oestradiol affect leptin receptor expression under physiological conditions. Radioactive in situ hybridization histochemistry was used to measure the gene expression under investigation. Total leptin receptor transcript levels were lowest in pro-oestrus in the choroid plexus, these changes correspond inversely with levels of circulating oestradiol in the rat 4-day oestrous cycle. In contrast full-length leptin receptor levels in both arcuate and ventromedial nuclei did not correspond to the levels of total leptin receptor in the same areas of the hypothalamus or serum levels of oestradiol. Full-length leptin receptor expression in the arcuate nucleus was negatively correlated with neuropeptide Y (NPY) expression (r = 0.447, p < 0.05) in the same nucleus. Arcuate nucleus NPY expression did not correlate significantly with the expression of total leptin receptors in the arcuate nucleus (r = 0.080) or serum leptin levels (r = 0.251). Our results demonstrate that leptin receptor expression is regulated during the oestrous cycle. The unchanged serum leptin levels during the oestrous cycle together with the correlation between the expression of leptin-RL and NPY provide circumstantial evidence that regulation of leptin receptor abundance in the hypothalamus governs the biological actions of leptin.
Cloning of Proopiomelanocortin from the Brain of the African Lungfish, Protopterus annectens, and the Brain of the Western Spadefoot Toad, Spea multiplicatus
Authors: Lee Jenny , Lecaude Stephanie , Danielson Phillip , Sollars Cristina , Alrubaian Jasem , Propper Catherine R. , Lihrmann Isabelle , Vaudry Hubert , Dores Robert M.
Published: April 2003

A degenerate primer, specific for the opioid core sequence YGGFM, was used to clone and sequence proopiomelanocortin (POMC) cDNAs from the brain of the African lungfish, Protopterus annectens, and from the brain of the western spadefoot toad, Spea multiplicatus. In addition, the opioid-specific primer was used to clone and sequence a 3′RACE product corresponding to a portion of the open reading frame of S. multiplicatus proenkephalin. For both species, cDNA was made from a single brain and a degenerate opioid-specific primer provided a reliable probe for detecting opioid-related cDNAs. The African lungfish POMC cDNA was 1,168 nucleotides in length, and contained regions that are similar to tetrapod POMCs and fish POMCs. The African lungfish POMC encodes a tetrapod-like γ-MSH sequence that is flanked by sets of paired basic amino acid proteolytic cleavage sites. The γ-MSH region in ray-finned fish POMCs either has degenerate cleavage sites or is totally absent in some species. However, the African lungfish γ-MSH sequence does contain a deletion which has not been observed in tetrapod γ-MSH sequences. The β-endorphin region of lungfish POMC has the di-amino acid sequence tryptophan-aspartic acid in the N-terminal region and an additional glutamic acid residue in the C-terminal region of β-endorphin – features found in fish β-endorphin, but not tetrapod β-endorphins. The western spadefoot toad POMC was 1,186 nucleotides in length, and exhibited an organizational scheme typical for tetrapod POMCs. However, the toad POMC did lack a paired basic amino acid proteolytic cleavage site N-terminal to the β-MSH sequence. Thus, like rat POMC, it is doubtful that β-MSH is an end product in either the toad brain or intermediate pituitary. At the amino acid level, the toad POMC had 76% sequence identity with Xenopus laevis POMC and 68% sequence identity with Rana ribidunda POMC. The use of these POMC sequences to assess phylogenetic relationships within anuran amphibians will be discussed. With respect to the fragment of S. multiplicatus proenkephalin cDNA, two metenkephalin sequences and the metenkephalin-RF sequence were found encoded in this fragment. As seen for X. laevis and R. ridibunda proenkephalin, a leuenkephalin sequence was not detected in the C-terminal region of the S. multiplicatus proenkephalin. The absence of a leuenkephalin sequence may be a common feature of anuran amphibian proenkephalins.
Differential Effects of Pregnancy on Mineralocorticoid and Glucocorticoid Receptor Availability and Immunoreactivity in Cortisol Feedback Sites
Authors: Roesch Darren M. , Keller-Wood Maureen
Published: April 2003

The suppression of plasma adrenocorticotropic hormone by very low levels of cortisol is reduced in pregnant adrenalectomized ewes, suggesting that pregnancy reduces the efficacy of the high-affinity corticosteroid receptor. This study was designed to determine the effects of pregnancy on the availability, immunoreactivity, and affinity of both corticosteroid receptors: the high-affinity mineralocorticoid receptor (MR) and the lower-affinity glucocorticoid receptor (GR). Availability was measured in the hypothalamus, pituitary, hippocampus and kidney using a saturation point radioligand binding assay. GR availability was significantly decreased in hippocampal cytosols obtained from pregnant ewes, but did not significantly change in other tissues. This finding is consistent with increased GR activation due to elevated circulating concentrations of cortisol. MR availability significantly increased from undetectable levels in hippocampal cytosols obtained from nonpregnant ewes to 2.8 ± 1.6 fmol/mg protein in pregnant ewes, suggesting a reduced MR activation in the hippocampus during pregnancy. MR availability tended to be greater in other tissues during pregnancy, but these differences were not significant. The amount of immunoreactive MR (iMR) and GR (iGR) protein was estimated by quantifying Western blots. iGR significantly increased in the pituitary, but did not significantly change in other tissues. In contrast, iMR was significantly increased during pregnancy in all tissues assayed, suggesting that an increased cytosolic MR protein amount contributes to the observed increase in MR availability. Since studies suggest that progesterone is a potent anticorticosteroid, we tested for evidence of endogenous inhibition of binding to MR and/or GR during pregnancy by determining MR and GR affinity in pituitary cytosols obtained from nonpregnant and pregnant ewes. Although there was a tendency towards a decreased affinity of the MR in pregnant ewes, there was no significant change in the KD of the pituitary MR or GR during pregnancy. We hypothesize that an alteration in activation and/or autoregulation of the MR during pregnancy, particularly in the hippocampus, may contribute to the observed changes in receptor availability and immunoreactivity and increase basal plasma cortisol levels during pregnancy.
Brain Potentials and Attention after Acute and Subchronic Intranasal Administration of ACTH 4–10 and Desacetyl-α-MSH in Humans
Authors: Smolnik Rüdiger , Mölle Matthias , Fehm Horst Lorenz , Born Jan
Published: April 2003

Neuropeptides related to adrenocorticotropin (ACTH) are potent regulators of neurobehavioral functions. In humans, ACTH and its behaviorally active fragment ACTH 4–10 have been consistently found to diminish event-related brain potential (ERP) signs of focussing attention. This study aimed at (1) evaluating effects of ACTH 4–10 on ERP indicators of attention in healthy controls after intranasal administration of the peptide. This route of administration has been proposed to provide a more direct access to the brain than the intravenous administration of the peptide, (2) comparing acute effects and effects of a subchronic treatment with ACTH 4–10, and (3) comparing effects of ACTH 4–10 with those of desacetyl-α-MSH (corresponding to ACTH 1–13 amide) which like ACTH 4–10 binds to subgroups of the melanocortin receptor family. Double-blind placebo-controlled experiments were completed in 54 healthy young subjects. ERPs were recorded while the subject performed an auditory selective attention task. Moreover, a modified Stroop interference test including motivational (food, sex) and nonmotivational words was performed. Acute intranasal administration of ACTH 4–10 (1 mg) reduced the processing negativity (PN) of the ERP over frontal and central cortical areas (p < 0.05) indicating diminished focussing of attention. Moreover, on this condition subjects were more prone to interference on the Stroop task especially with motivational words (p < 0.05). Subchronic administration of ACTH 4–10 (1 mg/day over 6 weeks) did not affect PN and Stroop performance. Acute intranasal administration of desacetyl-α-MSH at equimolar doses (1.68 mg) also remained ineffective. However, some measures of Stroop performance appeared to improve after subchronic desacetyl-α-MSH treatment. Results confirm an acute decrease in focussing of attention after ACTH 4–10. These effects of intranasal administration are likely to reflect a direct action of the peptide on respective brain functions. Moreover, they were specific to ACTH 4–10 and were not obtained after equimolar doses of desacetyl-α-MSH, thus excluding a mediation via the known melanocortin receptors.
Activation of Arcuate Nucleus Neurons by Systemic Administration of Leptin and Growth Hormone-Releasing Peptide-6 in Normal and Fasted Rats
Authors: Luckman Simon M. , Rosenzweig Ivana , Dickson Suzanne L.
Published: April 2003

Both leptin and growth hormone secretagogues are believed to have stimulatory effects on the hypothalamic growth hormone pulse generator, though whether these are achieved through the same pathway is unknown. Systemic administration of a normally maximal effective dose of the growth hormone secretagogue GHRP-6 to male rats causes the induction of c-Fos protein in the ventromedial aspect of the hypothalamic arcuate nucleus. The effect of the same dose of GHRP-6 is, however, much greater in animals that have been fasted for 48 h, suggesting that in the food-replete rat, arcuate neurons either show reduced sensitivity to endogenous growth hormone secretagogues or they are under the tonic inhibitory influences of other factors. The major populations of arcuate neurons activated by GHRP-6 have been shown to contain neuropeptide Y or growth hormone-releasing factor, while leptin is thought to be inhibitory to neuropeptide Y neurons. Leptin did not alter the response of the rats to GHRP-6. However, it was able by itself to induce c-Fos protein immunoreactivity in the ventral, including the ventrolateral, arcuate nucleus of fasted rats. This is a clear demonstration of the acute activation of arcuate neurons in the rat following systemic leptin injection and suggests that GHRP-6 and leptin act on the growth hormone axis via different pathways.
Effects of Androgens and Antiandrogens on the Quantitative Immunohistochemistry of Gonadotrope Cells in Prepubertal Male Rats
Authors: Cónsole Gloria M. , Rulli Susana B. , Riccillo Fernando L. , Calandra Ricardo S. , Gómez Dumm César L.A.
Published: April 2003

In the male rat, androgens are involved in the feedback regulation of gonadotropin synthesis and secretion. Specific androgen-receptor blockade by the nonsteroidal antiandrogens, flutamide and Casodex, has proven to be a valid tool for studying androgen effects in vivo. The aim of the present study was to investigate the effect of antiandrogen administration at the pituitary level by evaluating the changes in gonadotropes through quantitative immunohistochemistry, and by comparing these alterations with the effect of androgen deprivation by castration either with or without subsequent androgen replacement. Male Sprague-Dawley rats (23 days old) were randomly divided into 5 groups for the following treatments: (a) controls; (b) flutamide-injected (10 mg/rat/day in a gelatin vehicle); (c) Casodex-injected (10 mg/rat/day in an oil vehicle); (d) castrated, and (e) castrated and dihydrotestosterone propionate-replaced (40 µg/rat/day in an oil vehicle). Groups were then sacrificed after 10 days of maintenance under each condition. Pituitaries were fixed in Bouin’s fluid and embedded in paraffin. Serial sections (4 µm) were obtained at different levels and immunostained by means of the primary murine monoclonal antibodies anti-FSH and anti-LH and a peroxidase-mediated EnVision System (Dako). Measurements of volume density (VD) and individual mean cell area were made by means of an image-analysis system (Imaging Technology, Optimas). Serum FSH and LH levels were determined by radioimmunoassay (RIA). Serum gonadotropin levels, VD, and mean cell area increased significantly in the flutamide-treated, Casodex-treated, and castrated groups (p < 0.05). Androgen replacement in the castrated rats, however, reduced VD, mean cell area, and serum gonadotropins to levels comparable to those of controls. We conclude that either androgen blockade by antiandrogens or castration produce an enhancement in the gonadotrope cell population in prepubertal rats, as shown by an increase in both VD and mean cell area, as well as an elevation in FSH- and LH-immunoreactive cells. These observations correlate well with the changes found in the levels of circulating gonadotropins as measured by RIA.