[Respiratory physiotherapy in acute viral bronchiolitis in the newborn. Pro/con arguments]. / La kinésithérapie respiratoire dans la bronchiolite virale aiguë du nourrisson. Arguments pour/contre.

This article reports an exchange of unbiased arguments between Mr Guy Postiaux speaking in favour of respiratory physiotherapy in acute viral bronchiolitis in the newborn and Prof. Jean-Christoph Dubus arguing against. A review of the literature suggests that traditional methods of physiotherapy should be abandoned because they are not validated and because they have harmful side effects. The latest Cochrane revue (2016) suggests the use of slow expiration techniques that have some validated elements and cause no harmful side effects. Large multicentre studies should be undertaken to confirm or refute the results of the five studies in the Cochrane review. Their analysis would allow extraction of objective evidence for the efficacy of slow expiration techniques on the relief of bronchopulmonary obstruction and the reduction of the degree of severity in the short and medium term. Studies of the effect of ambulatory respiratory physiotherapy for bronchiolitis of a moderate degree not requiring hospitalisation are not available. An evaluation is needed which is based on the pathophysiology of multifactorial bronchial obstruction and on the physical signs, of which auscultation is the cornerstone.

Expression of RAS-like family members, c-jun and c-myc mRNA levels in neoplastic hemocytes of soft-shell clams Mya arenaria using microsphere-based 8-plex branched DNA assay.

The molecular mechanisms by which disseminated neoplasia (DN) is developed in soft shell clams Mya arenaria remain largely unknown. This study aims at quantifying Rho-like GTPase, RAS-Rho, RAS-related C3 botulinum (RAS C3), c-jun as well as c-myc transcript levels in clams sampled at North River (Charlottetown, Prince Edward Island, Canada). The transcripts were quantified using multiplex gene analysis (Quantigene(®) 2 Plex, Affymetrix) in 3 groups of clams: (1) Group C (healthy clams considered as control) with a low percentage of tetraploid hemocytes (<10%); (2) Group D (disease in development): individuals presenting a percentage of tetraploid cells ranging between 10% and 50%; (3) Group E (established disease): clams with a high percentage of tetraploid hemocytes (>50%). Data showed a down-regulation of Rho-like GTPase, Rho-like subfamily, RAS C3, c-jun and an up-regulation of c-myc gene expression. It is believed that a deregulation of the expression of these genes could partly unravel the molecular mechanisms involved in the development of DN in soft shell clams Mya arenaria. Further investigations should be pursued to determine the role of these gene products in clams' hemocytes.

Respiratory muscle endurance training in obese patients.

OBJECTIVE: Increased respiratory muscle work is associated with dyspnea and poor exercise tolerance in obese patients. We evaluated the effect of respiratory muscle endurance training (RMET) on respiratory muscle capacities, symptoms and exercise capacity in obese patients. DESIGN: A total of 20 obese patients hospitalized for 26 ± 6 days to follow a low-calorie diet and a physical activity program were included in this case-control study. Of them, 10 patients performed RMET (30-min isocapnic hyperpnea at 60-80% maximum voluntary ventilation, 3-4 times per week during the whole hospitalization period: RMET group), while the other 10 patients performed no respiratory training (control (CON) group). RMET and CON groups were matched for body mass index (BMI) (45 ± 7 kg m(-2)) and age (42 ± 12 years). Lung function, respiratory muscle strength and endurance, 6-min walking distance, dyspnea (Medical Research Council scale) and quality of life (short-form health survey 36 questionnaire) were assessed before and after intervention. RESULTS: Similar BMI reduction was observed after hospitalization in the RMET and CON groups (-2 ± 1 kg m(-2), P < 0.001). No significant change in lung function and respiratory muscle strength was observed except for vital capacity, which increased in the RMET group (+0.20 ± 0.26 l, P = 0.039). Respiratory muscle endurance increased in the RMET group only (+52 ± 27%, P < 0.001). Compared with the CON group, the RMET group had greater improvement in 6MWT (+54 ± 35 versus +1 ± 7 m, P = 0.007), dyspnea score (-2 ± 1 versus -1 ± 1 points, P = 0.047) and quality of life (total score: +251 ± 132 versus +84 ± 152 points, P = 0.018) after hospitalization. A significant correlation between the increase in respiratory muscle endurance and improvement in 6MWT distance was observed (r (2) = 0.36, P = 0.005). CONCLUSIONS: The present study indicates that RMET is feasible in obese patients and can induce significant improvement in dyspnea and exercise capacity. RMET may be a promising tool to improve functional capacity and adherence to physical activities in this population, but further studies are needed to confirm these results.

Receptor activated C kinase is down-regulated in the male gonad of the marine bivalve mollusc Mya arenaria exposed to tributyltin (TBT).

This study was aimed at investigating the molecular mechanisms by which tributyltin (TBT) impairs the reproductive processes in the marine bivalve Mya arenaria. The suppression polymerase chain reaction subtractive hybridization (SSH) method was used to identify differentially expressed transcripts in the gonads of adult M. arenaria 72 h after a single injection of 160 ng TBT in the adductor muscle. Subtractive cDNA libraries comprising 322 clones were obtained. These clones were sequenced and corresponded to 55 female and 26 single male non-redundant cDNAs. Following similarity searches in genome databases, some of the transcripts could be assigned to cellular functions including mitochondrial respiration, structural proteins, structure of cytoskeleton, nucleic acid regulation, general metabolism and signal transduction. Among the potentially differentially regulated transcripts, Receptor for activated C kinase 1 (RACK1) represented 6% of the total down-regulated clones in males and the corresponding protein exhibited a high degree of similarity (80%) with the human polypeptide. The RACK1 cDNA from M. arenaria consists of 1085 bp, encoding a 318 deduced polypeptide which contains five internal tryptophan-aspartate (WD) repeats, six putative PKC phosphorylation sites, one tyrosine kinase site, four putative N-myristoylation sites as well as a transmembrane segment spanning amino acid 228-251. A significant down-regulation (by approximately 30% (p<0.05)) of RACK1 expression in male gonads exposed to TBT was confirmed by quantitative real-time RT-PCR. Transcript levels of RACK1 were higher in the female gonads than in the mantle, gills and male gonads. Gene expression as detected by in situ hybridization was strong in mature oocytes comparatively to primary germ cells. RACK1 may be a useful biomarker for TBT exposure in the reproductive system of bivalve molluscs.

Molecular cloning of superoxide dismutase (Cu/Zn-SOD) from aquatic molluscs.

The potential of the first line of the active oxygen-scavenging system, partial cDNA encoding Cu/Zn superoxide dismutase (SOD) was isolated in three aquatic mollusc species: Ruditapes decussatus (marine clam), Dreissena polymorpha (continental water mussel) and Bathymodiolus azoricus (hydrothermal vent mussel). These SOD cDNA fragments were amplified by PCR with degenerate oligonucleotide primers derived from the amino acid sequence conserved in the Cu/Zn-SOD from several other organisms. A partial cDNA of CuZn-SOD was obtained for R. decussates (510 bp), D. polymorpha (510 bp) and B. azoricus (195 bp). The deduced amino acid sequence showed high similarity among the three mollusc species (57-63%) and among other species (50-65%). The residues involved in coordinating copper (His-47, 49, 64, 121) and zinc (His-64, 72, 81 and Asp-84) were well conserved among the three Cu/Zn-SOD sequences.

Identification of differentially expressed genes in Dreissena polymorpha exposed to contaminants.

Development of transcriptome analysis methods such as differential display PCR and construction of subtractive libraries now makes it possible to profile gene expression in response to xenobiotic exposure. As an example of application of these methods, zebra mussels (Dreissena polymorpha) were treated with various contaminants such as Aroclor 1254, 3-methylcholanthrene, chrysene and atrazine. A total of 242 mRNAs were identified as differentially expressed. Analysis of these mRNAs should provide valuable information regarding detoxification mechanisms in this bivalve species. In addition, the use of cDNA array technology applied to these gene products may constitute a multi-marker approach to monitor the effect of contamination on this aquatic species.

Quadrupolar transitions evidenced by resonant auger spectroscopy.

From absorption spectra, the only way to bring to the fore the occurrence of quadrupolar transitions is to study their angular dependence. Resonant spectroscopies offer a new opportunity to obtain more insight into excited electronic states by studying lineshape and intensity of decay processes. We show here that resonantly excited Ti KL(2,3)L(2,3) Auger spectra of TiO2(110) carry a clear signature of quadrupolar transitions to localized e(g) and t(2g) d-like states, giving access to a direct measurement of crystal field splitting.

Molecular cloning, characterization of cDNA, and distribution of mRNA encoding the frog prohormone convertase PC1.

Prohormone convertases (PCs) are calcium-dependent serine endoproteases of the subtilisin/kexin family that play a key role in the posttranslational processing of precursors for biologically active peptides. In this study, we have characterized the cDNA encoding PC1 in the European green frog Rana ridibunda. A frog brain cDNA library was screened by using a heterologous probe at low stringency, and a 2.3-kb cDNA clone encoding PC1 was isolated. This cDNA encodes a 736-residue protein with a 26-amino-acid signal peptide. Comparative structural analysis revealed that frog PC1 exhibits a high degree of amino acid identity with its mammalian counterparts, in particular in the subtilisin-like catalytic domain. Northern blot analysis resolved two major transcripts of 3.0 kb and 5.0 kb that were expressed differentially in the brain and pituitary. In situ hybridization studies showed that, in the frog brain, the highest densities of PC1 mRNA are present in the amygdala, the hypothalamus, and the anterior preoptic area. High concentrations of PC1 mRNA also were found in the pars distalis and pars intermedia of the pituitary, whereas the pars nervosa was devoid of hybridization signal. The wide distribution of PC1 mRNA in the brain and pituitary suggests that, in frog, PC1 is involved in the processing of a number of hormone and neuropeptide precursors.

Characterization of the cDNA encoding the prohormone convertase PC2 and localization of the mRNA in the brain of the frog Rana ridibunda.

A number of precursors for neuropeptides have recently been cloned in amphibians, but little is known concerning the endoproteases responsible for the processing of these precursors. Here we report on the molecular cloning of the cDNA encoding the proprotein convertase PC2 and the distribution of the corresponding mRNA in the European green frog Rana ridibunda. The full cDNA structure (2125 bp) was obtained from the analysis of the PCR products combined with the sequence from a clone isolated from a frog pituitary cDNA library. The deduced amino acid sequence revealed that frog PC2 comprises 636 amino acid residues including a 22-residue signal peptide. RT-PCR analysis showed that PC2 is expressed not only in the brain and pituitary but also in various peripheral organs including the pancreas, stomach, intestine, liver, kidney and testis. In situ hybridization histochemistry revealed that, in the central nervous system, PC2 mRNA is widely distributed, the highest concentrations being found in the pallium, the anterior preoptic area, the hypothalamus and the medial amygdala. High levels of PC2 mRNA were also detected in the intermediate lobe of the pituitary. The overall distribution of PC2 mRNA in the frog brain is consistent with its involvement in the processing of a number of neuropeptide and hormone precursors.

Structural organization of the gene encoding the neuroendocrine chaperone 7B2.

The neuroendocrine-specific polypeptide 7B2 is a constituent of the regulated secretary pathway. Recently, 7B2 was found to function as a molecular chaperone for prohormone convertase PC2. This report describes the genomic organization of the 7B2 gene which consists of six exons. Exon I corresponds to the 5'-untranslated mRNA region, while exons 2 and 3 encode the signal peptide and the amino-terminal half of the 7B2 protein that is distantly related to a subclass of molecular chaperones. The carboxy-terminal half of 7B2, responsible for its inhibitory action on PC2, is encoded by exons 4-6. Primer-extension analysis showed that the human 7B2 gene is transcribed from multiple transcription-initiation sites. The human 7B2 gene promoter contains a cAMP-responsive element, an AP-1 site, and several Pit-1/GHF-1-binding domains and heat-shock-element-like sequences but no obvious TATA or CAAT boxes. Of further interest is the finding of two DNA elements which are common to the promoter regions of the 7B2 gene and other genes selectively expressed in neuroendocrine tissues.

Short- and long-term effects of nucleus basalis magnocellularis lesions on cortical levels of somatostatin and its receptors in the rat.

Cognitive and histological alterations in human Alzheimer's disease (AD) are correlated with selective neuronal loss in nucleus basalis of Meynert. In search of an animal model of AD-linked neurochemical deficits, we examined the effects of short- (2 weeks) and long- (3 and 6 months) term lesions of the nucleus basalis magnocellularis (NBM) on somatostatinergic parameters in rat forebrain. NBM lesions were performed by unilateral injection of ibotenic acid into the NBM. Cortical choline-acetyl transferase (ChAT) activity and acetylcholinesterase staining in the NBM remained significantly decreased ipsi- as compared to contralaterally up to 6 months after the placement of the lesion. Somatostatin (SRIF) content was increased by 120% in the ipsilateral frontal cortex 6 months post-lesion but not at shorter time intervals. Levels of neuropeptide Y (which is extensively co-localized with SRIF in the forebrain) were not significantly altered after unilateral NBM lesions at any time point. A 30% decrease in SRIF binding capacity as well as a marked reduction of SRIF inhibition of adenylate cyclase, indicative of a loss of functional SRIF receptors, was observed in ipsilateral versus contralateral frontal cortex on brain tissue homogenates after short-term unilateral NBM lesion. By film radioautography, the loss in SRIF binding sites was localized to both superficial and deep layers of the frontal cortex. This loss persisted up to 3 months but was no longer apparent after 6 months due to a decrease in SRIF binding capacity on the contralateral side.

Neuropeptide-Y in the trout brain and pituitary: localization, characterization, and action on gonadotropin release.

Using a specific antiserum raised against synthetic neuropeptide-Y (NPY), the distribution of NPY-like immunoreactivity in the brain and pituitary of the trout Oncorhynchus mykiss has been examined with the indirect immunofluorescence and peroxidase-antiperoxidase methods. The highest density of NPY-immunoreactive elements was found in the basal telencephalon and hypothalamus. In particular, NPY-immunoreactive neurons were located in the nucleus entopeduncularis and the preoptic nucleus. NPY-immunoreactive fibers were observed throughout the trout brain. The preoptic nucleus, the suprachiasmatic nucleus, and the nucleus entopeduncularis were densely innervated. In addition, NPY-positive fibers were detected in the nucleus lateralis tuberis and in the distal and intermediate lobes of the pituitary. The NPY-like peptide of the trout brain was characterized by combining HPLC analysis and radioimmunological detection. Serial dilutions of trout hypothalamus and pituitary extracts produced displacement curves that were parallel to the standard curve. HPLC analysis resolved a major peak which was slightly less hydrophobic than porcine NPY. The possible effect of NPY, either alone or in combination with a GnRH antagonist, on gonadotropin (GtH) release from trout pituitaries was investigated using a perifusion system technique. Graded concentrations of synthetic NPY induced a dose-dependent stimulation of GtH release. The stimulatory activities of NPY and various short chain analogs on GtH release were compared: the order of potency was NPY greater than NPY-(2-36) greater than NPY-(16-36) greater than NPY-(25-36). This result suggests that the biological determinant of NPY is located in the C-terminal part of the molecule. Administration of a short pulse of NPY or GnRH (10(-7) M each) induced a marked stimulation of GtH release. Prolonged infusion of the GnRH antagonist D-Phe2-6,Pro3-GnRH induced a significant reduction of GnRH-evoked GtH secretion. In addition, the GnRH antagonist blocked NPY-induced GtH release. The widespread distribution of NPY in the trout brain suggests the involvement of this neuropeptide in a variety of physiological functions. The present data support the view that NPY, released by nerve terminals in the distal lobe of the pituitary, may act presynaptically on GnRH fibers to modulate GtH release.

Characterization of melanotropin-release-inhibiting factor (melanostatin) from frog brain: homology with human neuropeptide Y.

A polypeptide was purified from frog brain extracts on the basis of its ability to inhibit alpha-melanotropin release from perifused frog neurointermediate lobes. Based on Edman degradation, amino acid analysis, and peptide mapping, the primary structure of this frog melanotropin-release-inhibiting factor (melanostatin) was determined to be H-Tyr-Pro-Ser-Lys-Pro-Asp-Asn-Pro-Gly-Glu-Asp-Ala-Pro-Ala-Glu-Asp-Met- Ala-Lys-Tyr-Tyr-Ser-Ala-Leu-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg- Tyr-NH2 . Frog melanostatin belongs to the pancreatic polypeptide/neuropeptide Y/peptide YY family, and the structure of this peptide differs from that of human neuropeptide Y by only one amino acid substitution in position 19. A synthetic replicate of frog melanostatin is coeluted with the native peptide on HPLC and is highly potent in inhibiting alpha-melanotropin secretion in vitro (IC50 = 60 nM).

Localization and characterization of neuropeptide Y in the brain of Microcebus murinus (Primate, Lemurian).

The distribution of neuropeptide Y (NPY) in the brain of the lemur Microcebus murinus was determined by immunocytochemistry with the aid of a highly specific antiserum against synthetic porcine NPY. When compared with previous immunohistochemical data obtained in primates and other mammalian species, the localization of NPY-immunoreactive (IR) structures in the Microcebus murinus brain revealed particular features. (1) Numerous NPY-IR perikarya and a dense network of IR nerve terminals were found in the supraoptic and suprachiasmatic nuclei, respectively. The occurrence of NPY-IR perikarya in the supraoptic nucleus, also reported in the squirrel monkey, seems to be specific to primates. In the squirrel monkey, the suprachiasmatic nucleus exhibits only a moderate innervation, whereas in humans it appears totally devoid of NPY-IR fibers. (2) IR perikarya and axon processes were observed in many upper brainstem areas, in particular in the interpeduncular, raphe pontine, dorsal tegmental, parabrachial, and dorsal raphe nuclei, in the locus coeruleus, the nucleus of the solitary tract, and the reticular formation; in this latter area, the occurrence of two categories of NPY-IR neurons was demonstrated on the basis of their morphology and localization, suggesting that they may play distinct roles. (3) NPY-IR nerve processes could be traced over a long distance. (4) For the first time, numerous NPY-IR terminals were observed close to the lumen of the various cerebral ventricles. The immunoreactive NPY-like peptide was characterized by combining high performance liquid chromatography (HPLC) analysis and radioimmunoassay quantification. The dilution curves obtained with synthetic porcine NPY and serial dilutions of occipital cortex, paraventricular and supraoptic hypothalamus, posterior hypothalamus, medulla oblongata, or preoptic area extracts were parallel. The highest amounts of NPY were measured in the hypothalamus and telencephalon. HPLC analysis resolved a single peak of NPY-like immunoreactivity that exhibited the same retention time as synthetic porcine NPY. The distribution of NPY in the lemurian brain is discussed with respect to phylogeny and putative functions.

Neuropeptide Y: localization in the central nervous system and neuroendocrine functions.

Neuropeptide Y (NPY) is a 36-amino acid peptide first isolated and characterized from porcine brain extracts. A number of immunocytochemical investigations have been conducted to determine the localization of NPY-containing neurons in various animal species including both vertebrates and invertebrates. These studies have established the widespread distribution of NPY in the brain and in sympathetic neurons. In the rat brain, a high density of immunoreactive cell bodies and fibers is observed in the cortex, caudate putamen and hippocampus. In the diencephalon, NPY-containing perikarya are mainly located in the arcuate nucleus of the hypothalamus; numerous fibers innervate the paraventricular and suprachiasmatic nuclei of the hypothalamus, as well as the paraventricular nucleus of the thalamus and the periaqueductal gray. At the electron microscope level, using the pre- and post-embedding immunoperoxidase techniques, NPY-like immunoreactivity has been observed in neuronal cell body dendrites and axonal processes. In nerve terminals of the hypothalamus, the product of the immunoreaction is associated with large dense core vesicles. In lower vertebrates, including amphibians and fish, neurons originating from the diencephalic (or telencephalic) region innervate the intermediate lobe of the pituitary where a dense network of immunoreactive fibers has been detected. At the ultrastructural level, positive endings have been observed in direct contact with pituitary melanotrophs of frog and dogfish. These anatomical data suggest that NPY can act both as a neurotransmitter (or neuromodulator) and as a hypophysiotropic neurohormone. In the rat a few NPY-containing fibers are found in the internal zone of the median eminence and high concentrations of NPY-like immunoreactivity are detected in the hypothalamo-hypophyseal portal blood, suggesting that NPY may affect anterior pituitary hormone secretion. Intrajugular injection of NPY causes a marked inhibition of LH release but does not significantly affect other pituitary hormones. Passive immunoneutralization of endogenous NPY by specific NPY antibodies induces stimulation of LH release in female rats, suggesting that NPY could affect LH secretion at the pituitary level. However, NPY has no effect on LH release from cultured pituitary cells or hemipituitaries. In addition, autoradiographic studies show that sites for 125I-labeled Bolton-Hunter NPY or 125I-labeled PYY (2 specific ligands of NPY receptors) are not present in the adenohypophysis, while moderate concentrations of these binding sites are found in the neural lobe of the pituitary. It thus appears that the inhibitory effect of NPY on LH secretion must be mediated at the hypothalamic level.(ABSTRACT TRUNCATED AT 400 WORDS)

Neuropeptide Y inhibits thyrotropin-releasing hormone-induced stimulation of melanotropin release from the intermediate lobe of the frog pituitary.

Previous studies have shown that the release of melanotropin from frog neurointermediate lobes is under the control of two neuropeptides: thyrotropin-releasing hormone (TRH) stimulates, while neuropeptide Y (NPY) inhibits alpha-melanocyte-stimulating hormone (alpha-MSH) secretion from intact neurointermediate lobes in vitro. The aim of the present study was to investigate possible interactions between the two regulatory peptides at the pituitary level. Whole neurointermediate lobes or acutely dispersed pars intermedia cells from Rana ridibunda were perifused in vitro for 2 to 7.5 hr and the concentrations of alpha-MSH released into the effluent perifusate were monitored by radioimmunoassay. Administration of TRH (10(-7) M) or NPY (10(-7) M) to dispersed cells induced, respectively, marked stimulation or inhibition of alpha-MSH release. The effects of the two neuropeptides were similar to those observed using intact neurointermediate lobes, suggesting that TRH and NPY act directly on melanotropic cells. Perifused whole neurointermediate lobes were exposed to NPY (10(-8) to 3 x 10(-7) M) for 120 min and a single dose of TRH (10(-8) M) was administered during the prolonged infusion of NPY. Using this procedure, we observed a dose-dependent inhibition of TRH-evoked alpha-MSH release. These data support the concept that TRH and NPY act through a common intracellular pathway to regulate alpha-MSH release.

Studies on the mode of action of neuropeptide Y (NPY) on maturational gonadotropin (GtH) secretion from perifused rainbow trout pituitary glands.

The action of neuropeptide Y (NPY) and gonadotropin releasing hormone (s-GnRH) have been compared on the release of gonadotropin (GtH) by perifused rainbow trout pituitary glands sampled from freshly ovulated female rainbow trout. We have already demonstrated that NPY stimulated the secretion of GtH in vitro.The pituitary responses to s-GnRH and NPY were similar either after repeated 10 min infusions or a one hour prolonged application. In both cases, after the first application, the pituitary did not responded to subsequent secretagogues stimulations, and appeared to be desensitized. The stimulatory action of s-GnRH was partly inhibited (60%) by LH-RH antagonist (DpGlu(1), DPhe(2), DTrp(3-6)) LH-RH, which completely inhibited the response to NPY in perifused pituitary glands sampled from freshly ovulated females, but did not modify the response of pituitaries taken from vitellogenic animals in which NPY induced a transient inhibition of the GtH secretion. These results may indicate that the mode of action of NPY would differ between vitellogenic and matured animals. NPY also stimulated the GtH secretion from perifused pituitary dispersed cells prepared from pituitaries taken from freshly ovulated rainbow trout, indicating that NPY may act directly on the pituitary gonadotropic cells to stimulate GtH secretion.

Innervation of the pars intermedia and control of alpha-melanotropin secretion in the newt.

In this study we have investigated the presence of nerve fibers containing dopamine, thyrotropin-releasing hormone (TRH) and neuropeptide Y (NPY) in the pars intermedia of the crested newt and we have examined the possible effect of these neurohormones on the release of alpha-melanotropin (alpha-MSH) by neurointermediate lobes in vitro. By means of immunohistochemistry, we observed the presence of tyrosine hydroxylase (TH)-immunoreactive fibers in the pars intermedia of the crested newt. Using a specific antiserum to dopamine, these fibers appeared to be mainly dopaminergic in nature. Unlike anurans, urodele amphibians do not exhibit TRH or NPY-like immunoreactivity in the pars intermedia. A perifusion system technique for newt pituitaries was developed to investigate the effect of dopamine, TRH and NPY on alpha-MSH secretion. Administration of increasing concentrations of dopamine (from 10(-9) to 10(-5)M) induced a dose-related inhibition of alpha-MSH release. This inhibitory effect was mimicked by the dopamine agonist apomorphine (10(-6)M). In contrast, the secretory activity of the newt pars intermedia was not affected by administration of synthetic TRH or NPY (up to 10(-7) and 10(-6)M, respectively). These results indicate that the neurotransmitter dopamine likely plays a pivotal role in the regulation of melanotropin secretion in urodele amphibians.

Central-type benzodiazepines and the octadecaneuropeptide modulate the effects of GABA on the release of alpha-melanocyte-stimulating hormone from frog neurointermediate lobe in vitro.

The involvement of the GABA-benzodiazepine receptor complex in the regulation of melanotropin secretion has been investigated using perfused frog neurointermediate lobes. The GABAA agonist 3-amino-1 propane sulfonic acid mimicked the biphasic effect of GABA on alpha-melanocyte-stimulating hormone secretion: a brief stimulation followed by an inhibition of melanotropin secretion. The GABAA antagonist SR 95531 (10(-4) M) inhibited both stimulation and inhibition of alpha-melanocyte-stimulating hormone release induced by GABA (10(-4) M). Since the inhibitory effect of baclofen (10(-4) M) was partially antagonized by SR 95531 (10(-4) M), it appears that the GABAergic control of alpha-melanocyte-stimulating hormone release is mainly achieved through activation of GABAA receptors. GABA-induced stimulation of alpha-melanocyte-stimulating hormone release was inhibited by tetrodotoxin (10(-5) M), an Na+ -channel blocker, or nifedipine (10(-5) M), a voltage-dependent Ca2+ -channel blocker, suggesting that Na+ and Ca2+ ions are involved in the stimulatory phase of GABA action. Only central-type benzodiazepine binding site agonists such as clonazepam (10(-4) M) modified alpha-melanocyte-stimulating hormone release. In fact, clonazepam (10(-7) to 10(-5) M) led to a dose-dependent potentiation of both GABA-induced stimulation and inhibition of alpha-melanocyte-stimulating hormone release. This potentiating effect was antagonized by the GABAA antagonist SR 95531 (10(-4) M) or by the central-type benzodiazepine binding site antagonist flumazenil (10(-4) M), whereas picrotoxin (10(-4) M) abolished only the stimulatory phase.(ABSTRACT TRUNCATED AT 250 WORDS)