[Evaluation of the ovarian reserve]. / L'évaluation de la réserve ovarienne.

AIMS: This literature review attempts to answer the following questions: 1) When making an assessment of ovarian reserve (OR) in an infertile woman? and 2) what is the investigation that best identifies situations of low OR in order to guide patients towards appropriate care? RESULTS: Some elements of the clinical examination should prompt us to rapidly perform OR tests: patients over 35 years, with cycle disorders or a history of pelvic disease, chemotherapy or radiotherapy. The most widely used OR tests are FSH, estradiol, inhibin B, antral follicle count (AFC), ovarian volume, Antimüllerian hormone (AMH) and dynamic tests. Most studies evaluating these markers were performed within the frame of in vitro fertilization (IVF), and we have little information on their ability to predict spontaneous pregnancy. FSH seems an interesting marker to identify extreme situations of ovarian insufficiency when using a high threshold value. Estradiol assay must always be associated with it. The AFC and AMH both have a good ability to predict the response to ovarian stimulation in IVF. However, their association does not increase their strength of prediction. The ovarian volume, inhibin B as well as dynamic tests are not relevant markers. None of these markers can predict the chances of pregnancy in IVF. CONCLUSION: OR tests must be performed before assisted reproduction techniques (ART) and should include FSH, estradiol, and AMH or CFA. Each center must define its specific standards for the interpretation of results. This assessment will advise the couple about their chances of success in ART and the clinician to adapt its management.

Effect of chronic treatment with the antidepressant tianeptine on the hypothalamo-pituitary-adrenal axis.

The effects of acute and chronic administration of tianeptine, a novel antidepressant agent, on the hypothalamo-pituitary-adrenal axis were studied in the adult male rat. A single injection of tianeptine did not alter the activity of the hypothalamo-pituitary-adrenal axis. In contrast, chronic administration of tianeptine (10 mg/kg twice a day for 15 days) induced a significant decrease in the concentration of corticotropin-releasing factor (CRF) in the hypothalamus and adrenocorticotropin (ACTH) in the anterior lobe of the pituitary. Chronic tianeptine treatment did not modify CRF levels in the cerebral cortex and hippocampus, and did not alter alpha-melanocyte-stimulating hormone and beta-endorphin levels in the neurointermediate lobe of the pituitary. Using the in situ hybridization technique, we observed that chronic administration of tianeptine did not modify CRF mRNA levels in the paraventricular nucleus of the hypothalamus. The effect of chronic tianeptine treatment on the neuroendocrine response to stress was also investigated. Tube restraint stress for 30 min induced a significant depletion of hypothalamic CRF and a substantial increase of plasma ACTH and corticosterone. Tianeptine abolished the stress-induced reduction of hypothalamic CRF concentration and markedly reduced the stress-induced increase in plasma ACTH and corticosterone levels. Taken together, these results suggest that tianeptine acts primarily at the level of the hypothalamus: (1) in unstressed rats, tianeptine reduces hypothalamic CRF and pituitary ACTH contents; (2) in stressed animals, tianeptine attenuates the activation of the hypothalamo-pituitary-adrenal axis.

Purification and characterization of joining peptide and N-terminal peptide of proopiomelanocortin from the pars distalis of the bullfrog pituitary.

The joining peptide (JP) and the N-terminal peptide of proopiomelanocortin (NPP) were isolated from an acid-acetone extract of the distal lobe of the pituitary of the bullfrog, Rana catesbeiana, and purified by gel filtration and reverse-phase high performance liquid chromatography. The amino acid sequence of the bullfrog JP resembled the sequences of the JPs of Rana ridibunda (86% similarity) and Xenopus laevis (54% similarity), as deduced from the nucleotide sequences of their cDNAs. The amino acid sequence of bullfrog NPP showed 100%, 85%, and 50% similarity with those of Rana ridibunda, Xenopus laevis, and human NPPs, respectively. Administration of bullfrog NPP (0.05-5 micrograms/ml) to perifused Rana ridibunda interrenal slices induced a dose-dependent stimulation of corticosterone and aldosterone release. The present results indicate that the primary structure of NPP has been highly conserved during evolution. These data also reveal that NPP, which has no sequence homology with ACTH, exhibits a substantial corticotropic activity.

Glucocorticoids, transmitters and stress.

Many kinds of stress stimulate the neuroendocrine systems controlling catecholamine and glucocorticoid secretion. Stress-induced stimulation of CRF-containing neurons appears to be mediated by serotonergic, noradrenergic, and possibly other neuronal pathways. Stress can alter various neurobiological and endocrine functions, two essential components of the neuroendocrine responses being release of adrenalin from chromaffin cells of the adrenal medulla and secretion of glucocorticoids from adrenocortical cells. Activation of adrenal steroid secretion is mainly by a reflex activation of hypothalamic neurons, which stimulate ACTH secretion from the anterior pituitary. While the neuropeptide CRF plays a major role in the neuroendocrine response to stress, the neuronal signals which are responsible for the regulation of CRF neurons have not been completely elucidated. A number of other regulatory substances may also participate, alone or with CRF, in the control of ACTH secretion by pituitary corticotrophs, and there is increasing evidence that classical neurotransmitters or neuropeptides may act directly on adrenocortical cells to modulate corticosteroid secretion. We review the neuronal, neuroendocrine, and humoral pathways which participate in the regulation of stress-induced corticosteroid secretion, and present preliminary data on the effect of the tricyclic antidepressant, tianeptine in the response of the HPA axis to stress.

The novel antidepressant, tianeptine, reduces stress-evoked stimulation of the hypothalamo-pituitary-adrenal axis.

The possible effect of tianeptine, a novel antidepressant agent, on the neuroendocrine response to stress was investigated in adult male rats. Tube restraint stress for 30 min induced a marked increase of plasma ACTH and corticosterone. A single i.p. injection of tianeptine (10 mg/kg), 120 min before stress caused a significant decrease of ACTH and corticosterone levels. In order to investigate the kinetics of the effect of tianeptine, the drug was injected at various times (from 15 min to 12 h) before restraint stress. The inhibitory effect of tianeptine on stress-induced elevations of plasma ACTH and corticosterone occurred from 1 to 3 h after the injection. Administration of increasing doses of tianeptine revealed that only the highest doses (10 and 20 mg/kg) had a significant effect on stress-evoked stimulation of ACTH and corticosterone secretion. These results show that the antidepressant, tianeptine, reduces the activation of the hypothalamo-pituitary-adrenal (HPA) axis induced by restraint stress. Since depressed patients generally exhibit an elevated cortisol level, the present data suggest that part of the therapeutic properties of tianeptine could be accounted for by the effect of this antidepressant to modulate the activity of the HPA axis.

Central-type benzodiazepines inhibit release of alpha-melanocyte-stimulating hormone from the rat hypothalamus.

In a previous work, we have shown that GABA inhibits the release of alpha-melanocyte-stimulating hormone (alpha-melanotropin) from hypothalamic neurons through activation of GABAA receptors [Delbende et al. (1989) Brain Res. 497, 86-93]. Since GABA-gated channel activity can be allosterically modulated by a variety of compounds including benzodiazepines, we have investigated the effect of benzodiazepines in the control of alpha-melanotropin release by the rat basal hypothalamus. This study was conducted in vitro using perifused rat hypothalamic slices and the amount of alpha-melanotropin release was monitored with a sensitive and highly specific radioimmunoassay. Infusion of clonazepam (50 microM), a selective agonist for central-type benzodiazepine binding sites, induced an inhibition of KCl (50 mM)-evoked alpha-melanotropin release. The inhibitory effect of clonazepam was rapid and reversible. Administration of Ro 15-1788 (100 microM), a specific antagonist for central-type benzodiazepine receptors or SR 95531, a GABAA receptor antagonist, completely reversed the inhibitory effect of clonazepam. In addition, Ro 15-1788 and SR 95531 both enhanced the amplitude of the response observed during prolonged KCl infusion on alpha-melanotropin neurons, suggesting the existence of a tonic inhibitory effect of endogenous GABA and/or benzodiazepines in the release of alpha-melanotropin by hypothalamic neurons. To investigate further the effect of benzodiazepines in the regulation of alpha-melanotropin neurons, rats were treated in vivo with clonazepam (5 mg/kg) or the non-selective benzodiazepine receptor agonist diazepam (3 mg/kg). Both compounds caused a significant increase in the content of alpha-melanotropin and beta-endorphin in the rat hypothalamus within 3 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of ions and ionic channel activators or blockers on release of alpha-MSH from perifused rat hypothalamic slices.

The involvement of sodium and chloride ions in the process of alpha-melanocyte-stimulating hormone (a-MSH) release from hypothalamic neurons was investigated using perifused rat hypothalamic slices. Three different stimuli were found to increase a-MSH release from hypothalamic slices: high K+ concentration (50 mM), veratridine (50 microM), and the Na+/K(+)-ATPase inhibitor ouabain (1 mM). Spontaneous or K(+)-evoked a-MSH release was insensitive to the specific Na+ channel blocker tetrodotoxin (TTX; 1.5 microM) and to the blocker of K+ channels tetraethylammonium (TEA; 30 mM) or 4-aminopyridine (4-AP; 4 mM). In contrast, blockage of ouabain-sensitive Na+/K(+)-ATPase increased the resting level of a-MSH and caused a dramatic potentiation of K(+)-evoked a-MSH release. The Na+ channel activator veratridine (50 microM) triggered a-MSH release. This stimulatory effect was blocked by TTX and prolonged by TEA application, indicating the occurrence of voltage-sensitive Na+ and K+ channels on a-MSH neurons. Replacement of Na+ by impermeant choline ions from 95 to 60 mM did not alter K(+)-evoked a-MSH release. Conversely, dramatic reduction of the external Na+ concentration to 16 mM caused a robust increase of a-MSH secretion from hypothalamic neurons, likely through activation of the Na+/Ca2+ exchange system. These data indicate that the depolarizing effect of K+ results from direct activation of voltage-operated Ca2+ channels. The lack of effect of TEA on basal a-MSH release prompted us to investigate the possible involvement of chloride ions in the regulation of the spontaneous activity of a-MSH neurons. Substitution of Cl- for impermeant acetate ions did not affect basal or K(+)-evoked a-MSH release.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of alpha-melanocyte-stimulating hormone (alpha-MSH)-like peptides in discrete regions of the rat brain. In vitro release of alpha-MSH from perifused hypothalamus and amygdala.

The neuropeptide alpha-melanocyte-stimulating hormone (alpha-MSH) is synthesized by discrete populations of hypothalamic neurons which project in different brain regions including the cerebral cortex, hippocampus and amygdala nuclei. The purpose of the present study was to identify the alpha-MSH-immunoreactive species contained in these different structures and to compare the ionic mechanisms underlaying alpha-MSH release at the proximal and distal levels, i.e. within the hypothalamus and amygdala nuclei, respectively. The molecular forms of alpha-MSH-related peptides stored in discrete areas of the brain were characterized by combining high-performance liquid chromatography (HPLC) separation and radioimmunoassay detection. In mediobasal and dorsolateral hypothalamic extracts, HPLC analysis confirmed the existence of a major immunoreactive peak which co-eluted with the synthetic des-N alpha-acetyl alpha-MSH standard. In contrast, 3 distinct forms of immunoreactive alpha-MSH, which exhibited the same retention times as synthetic des-, mono- and di-acetyl alpha-MSH, were resolved in amygdala nuclei, hippocampus, cortex and medulla oblongata extracts. The proportions of acetylated alpha-MSH (authentic alpha-MSH plus diacetyl alpha-MSH) contained in these extrahypothalamic structures were, respectively, 78, 80, 60 and 92% of the total alpha-MSH immunoreactivity. In order to compare the ionic mechanisms underlaying alpha-MSH release from hypothalamic and extrahypothalamic tissues, we have investigated in vitro the secretion of alpha-MSH by perifused slices of hypothalamus and amygdala nuclei. High potassium concentrations induced a marked increase of alpha-MSH release from both tissue preparations. However, a higher concentration of KCl was required to obtain maximal stimulation of amygdala nuclei (90 mM) than hypothalamic tissue (50 mM).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of platelet-activating factor on hypothalamic and hypophyseal pro-opiomelanocortin-related peptides and hypothalamo-pituitary-adrenal axis in the rat.

To examine the effect of platelet-activating factor (PAF-acether) on pro-opiomelanocortin (POMC)-related peptides and on the hypothalamo-pituitary-adrenal axis, we administered PAF-acether and BN 52021, a selective PAF-acether antagonist, to freely moving rats. Minipumps loaded with either PAF-acether (30 micrograms/kg) or the vehicle alone were connected to the jugular vein for 7 days and positioned under the back skin of rats. A group of animals treated with PAF-acether also received 15 mg/kg of BN 52021 orally twice a day. In vivo treatment with PAF-acether alone or in association with BN 52021 did not affect the hypothalamic concentrations of corticotropin-releasing factor (CRF), alpha-melanocyte-stimulating hormone (alpha-MSH) and beta-endorphin. Using a perifusion system for rat hypothalamic slices, we did not observe any effect of PAF-acether on spontaneous or potassium-induced release of alpha-MSH in vitro. In addition, treatment of rats with PAF-acether alone or in association with BN 52021 did not modify the alpha-MSH or beta-endorphin concentration in the neurointermediate lobe of the pituitary. In contrast, in vivo administration of PAF-acether caused a significant reduction of ACTH concentration in the anterior lobe of the pituitary and a marked decrease in the corticosterone level in plasma and adrenal glands. The inhibitory effect of PAF-acether was reversed by concomitant administration of BN 52021. The ineffectiveness of PAF-acether to modulate in vitro ACTH release from perifused anterior pituitary fragments ruled out a direct effect of PAF-acether on corticotrophs. These findings support the view that PAF-acether exerts a specific inhibitory effect on the hypothalamo-pituitary-adrenal axis.

Proopiomelanocortin (POMC)-related peptides in the brain of the rainbow trout, Salmo gairdneri.

We have investigated the presence of ACTH, alpha-MSH and beta-endorphin, three peptides which derive from the multifunctional precursor protein proopiomelanocortin (POMC) in the brain of the rainbow trout Salmo gairdneri. Using both the indirect immunofluorescence and peroxidase-antiperoxidase techniques, a discrete group of positive cells was identified in the hypothalamus, within the anterior part of the nucleus lateralis tuberis. alpha-MSH-containing neurons represented the most abundant immunoreactive subpopulation. Coexistence of alpha-MSH, ACTH and beta-endorphin was observed in the lateral part of the nucleus. ACTH- and beta-endorphin-containing cells were mainly distributed in the rostral and caudal regions of the nucleus. In the medial portion of the nucleus lateralis tuberis, numerous cells were only stained for alpha-MSH. Moderate to dense plexuses of immunoreactive fibers were observed in the ventral thalamus and the floor of the hypothalamus. Some of these fibers projected towards the pituitary. The concentrations of ACTH, alpha-MSH and beta-endorphin-like immunoreactivities were measured in microdissected brain regions by means of specific radioimmunoassays. Diencephalon, mesencephalon and medulla oblongata extracts gave dilution curves which were parallel to standard curves. The highest concentrations of POMC-derived peptides were found in the diencephalon (alpha-MSH: 4.28 +/- 0.43 ng/mg prot.; ACTH: 1.08 +/- 0.09 ng/mg prot.; beta-endorphin: 1.02 +/- 0.1 ng/mg prot.), while lower concentrations were detected in the mesencephalon, medulla oblongata and telencephalon. The present results demonstrate that various peptides derived from POMC coexist within the same cell bodies of the fish hypothalamus. Taken together, these data suggest that expression and processing of POMC in the fish brain is similar to that occurring in pituitary melanotrophs.

gamma-Aminobutyric acid inhibits the release of alpha-melanocyte-stimulating hormone from rat hypothalamic slices.

The effect of gamma-aminobutyric acid (GABA) on release of alpha-melanocyte-stimulating hormone (alpha-MSH) from hypothalamic neurons was investigated in vitro using the perifusion technique. Rat hypothalamic slices were continuously superfused with Krebs-Ringer medium and the release of alpha-MSH in the effluent perifusate was monitored by means of a sensitive and specific radioimmunoassay method. Infusion of 50 mM K+ for 15 min induced a transient increase of alpha-MSH release (5- to 8-fold above the spontaneous level). Infusion of the same dose of K+ for 75 min caused a brief discharge of alpha-MSH during the first 30 min followed by sustained release of the neuropeptide. The effect of GABA was investigated 27 min after the onset of KCl infusion. Application of GABA (5 x 10(-5) M) resulted in a significant and reversible inhibition of K+-induced alpha-MSH release. The GABAA agonist, muscimol (10(-4) M), produced a prolonged inhibition of K+-evoked alpha-MSH release, while the GABAB agonist, baclofen (10(-4) M), was devoid of effect on hypothalamic alpha-MSH release. Bicuculline (10(-4) M), a specific GABAA antagonist, had no effect when added alone to the medium but totally reversed the inhibitory effect of GABA on K+-induced alpha-MSH release. Taken together, these data suggest that exogenous GABA exerts an inhibitory control on alpha-MSH neurons. Our data also show that the effect of GABA on alpha-MSH release by hypothalamic neurons is mediated through GABAA-type receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Involvement of voltage-operated calcium channels in alpha-melanocyte-stimulating hormone (alpha-MSH) release from perifused rat hypothalamic slices.

The contribution of voltage-operated calcium (VOC) channels in the mechanism of release of alpha-melanocyte-stimulating hormone (alpha-MSH) from hypothalamic neurons was investigated using perifused rat hypothalamic slices. The stimulatory effect of potassium (50 mM) on alpha-MSH release was completely blocked by cadmium (1 mM) a calcium competitor which indifferently blocks T-, L-and N-type VOC channels. To determine the nature of calcium conductances involved in K+-evoked alpha-MSH release, we have investigated the effect of a VOC channel agonist and 3 antagonists on the secretion of the neuropeptide. Administration of synthetic omega-conotoxin fraction GVIA (1 microM), a peptide toxin which blocks both N- and L-type VOC channels, reduced by 33% K+-induced alpha-MSH release. In contrast, the 1,4-dihydropyridine (DHP) antagonist nifedipine, at concentrations up to 100 microM, did not affect the response of hypothalamic alpha-MSH neurons to depolarizing concentrations of KCl. In addition, the secretion of alpha-MSH induced by high K+ concentrations was not reduced by nifedipine (10 microM) in the presence of diltiazem (1 microM), a benzothiazepine derivative which increases the affinity of the DHP antagonist for L-type VOC channels. The DHP agonist BAY K 8644 (0.1-10 microM) did not modify the early phase of the response of alpha-MSH neurons to K+-induced depolarization. In contrast BAY K 8644 (1 or 10 microM) significantly prolonged the duration of K+-induced alpha-MSH release. This sustained release of alpha-MSH induced by BAY K 8644 (10 microM) was totally suppressed by nifedipine (10 microM).(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of alpha-MSH-related peptides released from rat hypothalamic neurons in vitro.

Reverse-phase high-performance liquid chromatography analysis, coupled with a sensitive radioimmunoassay for alpha-melanocyte-stimulating hormone (alpha-MSH), was used to characterize the alpha-MSH-related peptides stored in the rat hypothalamus or released from perifused hypothalamic slices. Four peaks of alpha-MSH-like immunoreactivity (alpha-MSH-LI) co-eluting with synthetic des-N alpha-acetyl alpha-MSH, alpha-MSH and their respective sulfoxide derivatives were resolved and quantified. In hypothalamic extract, deacetyl alpha-MSH which was the predominant peptide represented 94.4% of total alpha-MSH-LI content, while the relative amount of alpha-MSH was only 5.6%. Analysis of alpha-MSH-related peptides contained in effluent perifusates showed that deacetyl alpha-MSH and its oxidized form were the major peptides released from neurons in basal conditions or under KCl-induced depolarization (50 mM KCl for 75 min). However, the proportion of acetylated peptide was 3-4 times higher in the perifusion medium than in hypothalamic extracts. Our data indicate that acetylation of des-N alpha-acetyl alpha-MSH may occur during the process of exocytosis. Since acetylation of alpha-MSH markedly increases the behavioural potency of the peptide, these results suggest that regulation of the acetyltransferase activity could be a key mechanism to modulate the bioactivity of alpha-MSH-related peptides in the brain.

Melanin-concentrating hormone (MCH) immunoreactivity in the brain and pituitary of the dogfish Scyliorhinus canicula. Colocalization with alpha-melanocyte-stimulating hormone (alpha-MSH) in hypothalamic neurons.

The distribution of melanin-concentrating hormone (MCH) in the central nervous system of the dogfish Scyliorhinus canicula was determined by indirect immunofluorescence and peroxidase-anti-peroxidase techniques, using an antiserum raised against synthetic salmon MCH. Three groups of MCH-positive cell bodies were localized in the posterior hypothalamus. The most prominent cell group was detected in the nucleus sacci vasculosi. Scattered MCH-immunoreactive cells were observed in the nucleus tuberculi posterioris and in the nucleus lateralis tuberis. At the pituitary level, the caudal part of the median lobe of the pars distalis contained strongly MCH-positive perikarya. Some of these cells were liquor-contacting-type. Immunoreactive fibers originating from the hypothalamic perikarya projected throughout the dorsal wall of the posterior hypothalamus. Positive fibers were also detected within the thalamus and the central gray of the mesencephalon. The distribution of MCH-containing neurons was compared to that of alpha-MSH-immunoreactive elements using consecutive, 5-micron thick sections. Both MCH- and alpha-MSH-immunoreactive peptides were found in the same neurons of the nucleus sacci vasculosi. These data suggest that MCH and alpha-MSH, two neuropeptides which exert antagonistic activities on skin melanophores, may also act in a coordinate manner in the central nervous system of cartilaginous fish.

Immunocytochemical Localization and Biochemical Characterization of alpha-Melanocyte-Stimulating Hormon in the Brain of the Rainbow Trout, Salmo gairdneri.

Abstract The distribution of alpha-melanocyte-stimulating hormone (alpha-MSH)-like immunoreactivity in the central nervous system of the rainbow trout Salmo gairdneri was investigated by indirect immunofluorescence and peroxidase-antiperoxidase techniques, using a highly specific antiserum generated in rabbits against synthetic alpha-MSH. Immunoreactive perikarya were exclusively observed in the basal hypothalamus within the pars anterioris of the nucleus lateralis tuberis. In this region, a moderate number of small stained cell bodies were observed surrounding the dorsal wall of the anterior infundibular recess. These immunoreactive cells were organized in rostro-caudal rows extending over the whole portion of the nucleus. Positive fibres originating from these perikarya were visualized in the dorsal posterior lobe and the ventral hypothalamus. A dense tract of immunoreactive fibres projected ventrally through the pituitary stalk and terminated in the neurohypophysis. The concentrations of alpha-MSH in different regions of the brain were measured by means of a sensitive and specific radioimmunoassay. The dilution curves obtained with synthetic alpha-MSH and serial dilutions of diencephalon, mesencephalon, medulla oblongata, telencephalon or pituitary extracts were strictly parallel. The highest concentration of alpha-MSH in brain was found in the diencephalon (1.31 +/- 0.07 ng/mg protein). In contrast alpha-MSH was not detectable in cerebellar extracts. Reverse-phase high-performance liquid chromatography and radioimmunoassay were used to characterize alpha-MSH-like peptides in the trout brain and pituitary. Two major forms of immunoreactive alpha-MSH were resolved by high performance liquid chromatography in hypothalamic extracts; these peptides exhibited the same retention times as des-Na-acetyl alpha-MSH and its sulfoxide derivative, respectively. Additional peaks of alpha-MSH immunoreactive material were detected in pituitary extract. These latter peptides coeluted with authentic alpha-MSH, diacetyl alpha-MSH and their sulfoxide forms. These results provide the first evidence for the presence of alpha-MSH in the brain of a teleostean fish. Our data indicate that, in the brain, the immunoreactivity corresponds to the non-acetylated form of alpha-MSH, while three different types of alpha-MSH-like molecules (namely deacetylated, monoacetylated, and diacetylated forms) coexist in the pituitary. It thus appears that, in salmonoid fish, mono- or diacetylation of the N-terminal serine residue of aL-MSH only occurs at the pituitary level.

Regional distribution of vasoactive intestinal peptide in brains from normal and parkinsonian subjects.

The distribution of vasoactive intestinal peptide (VIP) in the post-mortem human brain was determined by radioimmunoassay using a highly specific antiserum. The detection limit of the assay was 4 fmol/tube. The highest concentrations of VIP were found in the cerebral cortex, amygdala, hypothalamus and hippocampus. The lowest levels of peptide were detected in basal ganglia including caudate nucleus, external pallidum, putamen and substantia nigra. All dilution curves of acetic acid extracts from different brain areas were strictly parallel to the standard curve. Sephadex G-50 gel filtration of frontal cortex extract showed that VIP-like immunoreactivity (VIP-LI) eluted as a major peak comigrating with synthetic hVIP. Detailed mapping of VIP in the human cerebral cortex showed the existence of a rostro-caudal gradient of VIP-LI concentrations: the frontal cortex exhibited the highest VIP levels, the parietal and temporal cortex contained medium values and the occipital cortex contained the lowest VIP levels. The concentrations of VIP-LI were compared in various regions of the human brain from normal and parkinsonian subjects. No significant changes in VIP-LI levels occurred in the brains of patients dying with Parkinson's disease. No difference in VIP levels could be found either when the parkinsonian group was subdivided into nondemented and demented patients. These data indicate that VIP-containing neurons are not affected in parkinsonian patients. Our results also suggest that VIP neuronal systems are not involved in the course of dementing process in Parkinson's disease.

Alpha-melanocyte-stimulating hormone (alpha-MSH) in the brain of the cartilagenous fish. Immunohistochemical localization and biochemical characterization.

The distribution of immunoreactive alpha-melanocyte-stimulating hormone (alpha-MSH) in the central nervous system and pituitary of the elasmobranch fish Scyliorhinus canicula was determined by the indirect immunofluorescence and the peroxidase-antiperoxidase methods using a highly specific antiserum. Perikarya containing alpha-MSH-like immunoreactivity were localized in the dorsal portion of the posterior hypothalamus, mainly in the tuberculus posterioris and sacci vasculosus nuclei. Immunoreactive alpha-MSH cell bodies were found in the dorsal wall and ventral region of the caudal part of the tuberculum posterioris. These structures were densely innervated by fine beaded immunoreactive fibers. Some alpha-MSH immunoreactive cells were occasionally detected in the ventral part of the nucleus periventricularis. Scattered cell bodies and fibers were also observed in the dorsal wall of the posterior recess. Outside the hypothalamus very few fibers were detected in the dorsal thalamus and mesencephalon. No immunoreactivity was found in any other parts of the brain. The alpha-MSH immunoreactive material localized in the brain was characterized by combining high-performance liquid chromatography (HPLC) analysis and radioimmunological detection. Brain and pituitary extracts exhibited displacement curves which were parallel to that obtained with synthetic alpha-MSH. The concentrations of alpha-MSH immunoreactive material were determined in 5 different regions of the brain. The highest concentration was found in the hypothalamus. HPLC analysis resolved two major forms of immunoreactive alpha-MSH in the hypothalamus, which had been same retention times as des-N alpha-acetyl-alpha-MSH and its sulfoxide derivative. These results provide the first evidence for the presence of alpha-MSH-like peptides in the fish brain.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypothalamic alpha-melanocyte-stimulating hormone (alpha-MSH) is not under dopaminergic control.

A possible dopaminergic regulation of hypothalamic proopiomelanocortin (POMC)-containing neurons has been investigated in rats by means of in vivo and in vitro approaches. Acute or 3-weeks chronic in vivo treatments with the dopaminergic agonists apomorphine (1 mg/kg: s.c.) and 2-Br-alpha-ergocriptine (2.5 mg/kg; s.c.) or the dopaminergic antagonist haloperidol (0.15-3 mg/kg; i.p.) had no significant effect on the concentration of alpha-melanocyte-stimulating hormone (alpha-MSH) in two hypothalamic regions: arcuate nucleus (AN) and dorsolateral area (DLH). In the same way, chronic administration of the dopaminergic agonists or antagonist did not induce any change in hypothalamic contents of beta-endorphin, another peptide derived from POMC. Reverse-phase high-performance liquid chromatographic analysis revealed that acetic acid extracts of AN and DLH both contained two major forms of alpha-MSH-like peptides: deacetylated alpha-MSH and authentic alpha-MSH. The ratio between these two forms was not altered after acute haloperidol treatment (3 mg/kg, i.p.). The possible effect of dopamine on the release of hypothalamic alpha-MSH was studied in vitro using perifused rat hypothalamic slices. Infusion of dopamine (10(-7)-10(-5)M) or its antagonist haloperidol (10(-5)M) had no effect on spontaneous alpha-MSH release from hypothalamic tissue. In addition, none of these drugs had any effect on potassium (50 mM)-induced alpha-MSH release. It is concluded that dopaminergic neurons are not involved in the regulation of synthesis, post-translational processing (acetylation) or release of hypothalamic alpha-MSH.