Biomedical subjects

H Vaudry

Publications and source records attributed to H Vaudry.

At least 109 records · Page 6Linked to original sources

PubMed2000

Rat glomerulosa cells express functional 5-HT7 receptors.

It has been previously demonstrated that serotonin (5-HT) is a potent stimulator of aldosterone secretion in amphibians and mammals. The aim of the present study was to characterize the type of serotonergic receptor involved in the action of 5-HT on rat glomerulosa cells. The effects of 10 serotonergic receptor agonists and 12 receptor antagonists on aldosterone secretion from perifused rat adrenocortical slices were investigated. Correlation analysis between the potencies of the different compounds in our model and those previously reported for transfected 5-HT receptors showed that the rat adrenal 5-HT receptor exhibits the pharmacological profile of the 5-HT7 receptor. RT-PCR amplification with specific primers for the 5-HT7 receptor confirmed the presence of 5-HT7 receptor mRNA in the adrenal cortex. Western blot analysis using antibodies against the 5-HT7 receptor revealed the occurrence of a protein with an apparent molecular mass of 66 kDa in the zona glomerulosa. In glomerulosa cells, 5-HT induced a concentration-dependent increase of cAMP formation. These data demonstrate that rat adrenal glomerulosa cells express functional 5-HT7 receptors positively coupled to adenylyl cyclase.

Aldosterone↗

PubMed2000

Pituitary adenylate cyclase-activating polypeptide and its receptors: from structure to functions.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a 38-amino acid peptide that was first isolated from ovine hypothalamic extracts on the basis of its ability to stimulate cAMP formation in anterior pituitary cells. PACAP belongs to the vasoactive intestinal polypeptide (VIP)-glucagon-growth hormone releasing factor-secretin superfamily. The sequence of PACAP has been remarkably well conserved during the evolution from protochordate to mammals, suggesting that PACAP is involved in the regulation of important biological functions. PACAP is widely distributed in the brain and peripheral organs, notably in the endocrine pancreas, gonads, and respiratory and urogenital tracts. Characterization of the PACAP precursor has revealed the existence of a PACAP-related peptide whose activity remains unknown. Two types of PACAP binding sites have been characterized. Type I binding sites exhibit a high affinity for PACAP and a much lower affinity for VIP whereas type II binding sites have similar affinity for PACAP and VIP. Molecular cloning of PACAP receptors has shown the existence of three distinct receptor subtypes, the PACAP-specific PAC1 receptor, which is coupled to several transduction systems, and the two PACAP/VIP-indifferent VPAC1 and VPAC2 receptors, which are primarily coupled to adenylyl cyclase. PAC1 receptors are particularly abundant in the brain and pituitary and adrenal glands whereas VPAC receptors are expressed mainly in the lung, liver, and testis. The wide distribution of PACAP and PACAP receptors has led to an explosion of studies aimed at determining the pharmacological effects and biological functions of the peptide. This report reviews the current knowledge concerning the multiple actions of PACAP in the central nervous system and in various peripheral organs including the endocrine glands, the airways, and the cardiovascular and immune systems, as well as the different effects of PACAP on a number of tumor cell types.

Amino Acid Sequence↗

PubMed2000

Role of 5-HT in the regulation of the brain-pituitary-adrenal axis: effects of 5-HT on adrenocortical cells.

Serotonin (5-HT) plays a pivotal role in the regulation of the brain-pituitary-adrenal axis. In particular, 5-HT has been shown to control the activity of hypothalamic CRF neurons and pituitary corticotrope cells through activation of 5-HT1A and (or) 5-HT(2A/2C) receptor subtypes. 5-HT, acting through 5-HT2 receptors, can also trigger the renin-angiotensin system by stimulating renin secretion and consequently can enhance aldosterone production. At the adrenal level, 5-HT produced locally stimulates the secretory activity of adrenocortical cells through a paracrine mode of communication. The presence of 5-HT in the adrenal gland has been demonstrated immunohistochemically and biochemically in various species. In the frog, rat, and pig adrenal gland, 5-HT is synthesized by chromaffin cells, while in the mouse adrenal cortex, 5-HT is contained in nerve fibers. In man, 5-HT is present in perivascular mast cells. In vivo and in vitro studies have shown that 5-HT stimulates corticosteroid secretion in various species (including human). The type of receptor involved in the mechanism of action of 5-HT differs between the various species. In frogs and humans, the stimulatory effect of 5-HT on adrenocortical cells is mediated through a 5-HT4 receptor subtype positively coupled to adenylyl cyclase and calcium influx. In the rat, the effect of 5-HT on aldosterone secretion is mediated via activation of 5-HT7 receptors. Clinical studies indicate that 5-HT4 receptor agonists stimulate aldosterone secretion in healthy volunteers and in patients with corticotropic insufficiency and primary hyperaldosteronism. Local serotonergic control of corticosteroid production may be involved in the physiological control of the activity of the adrenal cortex as well as in the pathophysiology of cortisol and aldosterone disorders.

Adrenal Cortex↗

PubMed1999

Synergistic action of upstream elements and a promoter-proximal CRE is required for neuroendocrine cell-specific expression and second-messenger regulation of the gene encoding the human secretory protein secretogranin II.

Secretogranin II (SgII) is a secretory polypeptide stored in large dense core vesicles of neuroendocrine and neuronal cells. In order to characterize the molecular mechanisms underlying the tissue-specific expression of the SgII gene and its regulation by second-messenger pathways in endocrine and neuronal cells, we have cloned and characterized the human SgII gene. Sequence analysis revealed the existence of numerous putative cis-regulatory elements in the SgII gene promoter, including a consensus cyclic AMP-responsive element (CRE). Constructs containing different portions of the human SgII promoter fused to the luciferase reporter were transfected in AtT-20, SH-SY5Y, LLC-PK1 or COS-7 cells. Northern blot analysis showed that the endogenous SgII gene is more highly expressed in AtT-20 cells than in SH-SY5Y cells, and not expressed at all in LLC-PK1 cells. Treatment by forskolin or 12-O-tetradecanoylphorbol-13-acetate (TPA) caused a 1.5- and 10-fold increase, respectively, in SgII mRNA levels in SH-SY5Y cells but not in AtT-20 cells. Transfection experiments revealed that 4 kb of the human SgII promoter is sufficient to impart cell-specific expression of the reporter gene in the four cell lines studied. Specifically, in AtT-20 cells, a positive element located between -1.38 and -4 kb, in addition to the CRE, is responsible for the high expression of the SgII gene. In SH-SY5Y cells, a negative element located between -0.66 and -1.4 kb represses the activating effect of the CRE leading to an overall lower activity of fusion genes in these cells compared to the activity in AtT-20 cells. Finally, the promoter activity was very low in LLC-PK1 and COS-7 cells. Forskolin and TPA stimulated the activity of a SgII-luciferase fusion gene in SH-SY5Y but not in AtT-20 cells. Disruption of the CRE abolished the stimulatory effect of forskolin and TPA. These data suggest that the basal activity of the human SgII gene relies on cell-specific trans-acting factors in addition to factors that bind to the CRE and show that the regulation of this gene by second messengers is cell-specific and requires an intact CRE.

Base Sequence↗

PubMed1999

The pituitary-skin connection in amphibians. Reciprocal regulation of melanotrope cells and dermal melanocytes.

In amphibians, alpha-MSH secreted by the pars intermedia of the pituitary plays a pivotal role in the process of skin color adaptation. Reciprocally, the skin of amphibians contains a number of regulatory peptides, some of which have been found to regulate the activity of pituitary melanotrope cells. In particular, the skin of certain species of amphibians harbours considerable amounts of thyrotropin-releasing hormone, a highly potent stimulator of alpha-MSH release. Recently, we have isolated and sequenced from the skin of the frog Phyllomedusa bicolor--a novel peptide named skin peptide tyrosine tyrosine (SPYY), which exhibits 94% similarity with PYY from the frog Rana ridibunda. For concentrations ranging from 5 x 10(-10) to 10(-7) M, SPYY induces a dose-related inhibition of alpha-MSH secretion. At a dose of 10(-7) M, SPYY totally abolished alpha-MSH release. These data strongly suggest the existence of a regulatory loop between the pars intermedia of the pituitary and the skin in amphibians.

Amino Acid Sequence↗

PubMed1999

Characterization and localization of pituitary adenylate cyclase-activating polypeptide (PACAP) binding sites in the brain of the frog Rana ridibunda.

The biochemical characteristics and the distribution of pituitary adenylate cyclase-activating polypeptide (PACAP) binding sites have been investigated in the brain of the frog Rana ridibunda by using [(125)I]PACAP27 as a radioligand. Membrane-binding studies revealed the existence of high-affinity receptors for frog PACAP38 and PACAP27. In contrast, the [Des-His(1)]PACAP38 analogue had a much lower affinity and vasoactive intestinal polypeptide did not produce any displacement of the binding. Autoradiographic labeling of frozen brain sections revealed that the highest concentrations of PACAP receptors were located in the olfactory bulb, pallium, striatum, habenular nuclei, ventromedial thalamic nucleus, corpus geniculatum, posterior tubercle, dorsal part of the magnocellular preoptic nucleus, tectum, and the molecular cell layer of the cerebellum. Moderate binding was observed in the septum, in most parts of the thalamus, the dorsal hypothalamic nucleus, the median eminence, the ventral nuclei of the tegmentum, the torus semicircularis, and the interpeduncular and isthmi nuclei. The present data provide the first biochemical characterization and anatomic distribution of PACAP binding sites in the brain of a nonmammalian vertebrate species. The widespread distribution of specific PACAP receptors in the frog brain suggests that the peptide does not act solely as a hypophysiotropic factor, but likely fulfills neurotransmitter functions, neuromodulator functions, or both.

Animals↗

PubMed1999

Cloning, sequence analysis and tissue distribution of the mouse and rat urotensin II precursors.

Urotensin II (UII) is a cyclic neuropeptide initially isolated from the caudal neurosecretory system of teleost fish. The recent cloning of the UII precursor in frog and human has demonstrated that the peptide is not restricted to the fish urophysis but that it is also expressed in the central nervous system of tetrapods. Here, we describe the characterization of the cDNAs encoding prepro-UII in mouse and rat. A comparison of the primary structures of mouse and rat UII with those of other vertebrate UII reveals that the sequence of the cyclic region of the molecule (CFWKYC) has been fully conserved. In contrast, the N-terminal flanking domain of prepro-UII has markedly diverged with only 48% sequence identity between the mouse or rat and the human precursors. In situ hybridization histochemistry showed that the prepro-UII gene is predominantly expressed in motoneurons of the brainstem and spinal cord, suggesting that UII may play a role in the control of neuromuscular functions.

Amino Acid Sequence↗

PubMed1999

Molecular cloning of the cDNAs and distribution of the mRNAs encoding two somatostatin precursors in the African lungfish Protopterus annectens.

The occurrence of two somatostatin precursors, PSS1 and PSS2, yielding S-14 (SS1) and the variant [Pro2, Met13]S-14 (SS2), has been recently reported in the frog Rana ridibunda. The evolutionary significance of frog PSS2 is unclear because its sequence exhibits very little similarity with other known vertebrate somatostatin precursors. In the present study, we report on the characterization of two somatostatin precursor cDNAs from the brain of the African lungfish Protopterus annectens. One of the cDNAs encodes a 115-amino-acid protein that contains the SS1 sequence at its C-terminal extremity and thus is clearly homologous to PSS1. Comparison with other vertebrate PSS1 showed that lungfish PSS1 is more closely related to PSS1 from tetrapods than to PSS1 from fish. The other cDNA encodes a 109-amino-acid protein that contains a somatostatin variant [Pro2]S-14 at its C-terminal extremity. Sequence analysis of this second precursor indicated that it is the lungfish counterpart of frog PSS2. Northern blot analysis showed that lungfish PSS1 mRNA is widely distributed in the central nervous system and in peripheral organs, including the pancreas and gastrointestinal tract. In contrast, PSS2 mRNA was primarily found in the central nervous system but not in the pancreas or gut. In situ hybridization studies showed that the two genes are differentially expressed in various regions of the lungfish brain. The present data indicate that the PSS2 gene, initially discovered in frog, appeared early in vertebrate evolution, before the emergence of the tetrapod lineage. The recent isolation of a [Pro2]S-14 variant in the sturgeon, whose sequence is identical to that of lungfish SS2, suggests that the PSS2 gene may actually be present in the genome of all Osteichthyii.

Amino Acid Sequence↗

PubMed1999

Neurotrophic activity of pituitary adenylate cyclase-activating polypeptide on rat cerebellar cortex during development.

High concentrations of pituitary adenylate cyclase-activating polypeptide (PACAP) receptors are present in the external granule cell layer of the rat cerebellum during postnatal development. In vitro studies have shown that PACAP promotes cell survival and neurite outgrowth on immature cerebellar granule cells in primary culture. In the present study, we have investigated the effect of PACAP on the development of the cerebellar cortex of 8-day-old rats. Incubation of cultured granule cells for 12 or 18 h with PACAP provoked a significant increase in the rate of incorporation of [(3)H]thymidine in cultured granule cells, suggesting that PACAP could stimulate the proliferation of granule cells. After 96 h of treatment, in vivo administration of PACAP provoked a transient increase in the number of granule cells in the molecular layer and in the internal granule cell layer. In contrast, PACAP did not affect the number of Purkinje cells. The augmentation of the number of granule cells evoked by PACAP was significantly inhibited by the PACAP receptor antagonist PACAP(6-38). Administration of PACAP also caused a significant increase in the volume of the cerebellar cortex. The present study provides evidence that PACAP can act in vivo as a trophic factor during rat brain development. Our data indicate that PACAP increases proliferation and/or inhibits programmed cell death of granule cells, as well as stimulating neuronal migration from the external granule cell layer toward the internal granule cell layer.

Aging↗

PubMed1999

Protective effects of preconditioning in cultured rat endothelial cells: effects on neutrophil adhesion and expression of ICAM-1 after anoxia and reoxygenation.

BACKGROUND: Preconditioning with brief periods of ischemia protects the coronary endothelium against acute and chronic reperfusion injury, but the mechanisms of this endothelial protection remain unknown. We hypothesized that preconditioning protects endothelial cells through a decreased production of endothelial adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1), leading to a lesser adhesion of neutrophils to the endothelium. METHODS AND RESULTS: Cultured rat aortic endothelial cells were subjected to 6-hour anoxia followed by various durations of reoxygenation. Preconditioning was induced by 1-hour anoxia and 1-hour reoxygenation. ICAM-1 gene expression was measured by polymerase chain reaction, and the percentage of cells expressing ICAM-1 was assessed by confocal laser fluorescence microscopy. Anoxia/reoxygenation increased expression of ICAM-1, with a peak occurring after 6 hours of reoxygenation for mRNA and 9 hours for protein. Preconditioning prevented the increase in ICAM-1. Similar reductions were observed with the free radical scavenger N-2 mercaptopropionyl glycine (MPG). The inhibitory effect of preconditioning on ICAM-1 expression was abolished by an inhibitor of protein kinase C, an inhibitor of nitric oxide synthesis, and by MPG but was not affected by an adenosine receptor antagonist. Finally, both preconditioning and MPG partially prevented the increased adhesion of human neutrophils to reoxygenated endothelial cells. CONCLUSIONS: Preconditioning prevented reoxygenation-induced, free radical-mediated expression of ICAM-1 by a mechanism involving activation of protein kinase C and production of nitric oxide and free radicals, and this is associated with a lesser adhesion of neutrophils to endothelial cells. Such prevention of neutrophil adhesion may contribute to the protective effect of preconditioning against reperfusion-induced endothelial injury.

Animals↗

PubMed1999

Mu-opioid receptor mRNA expression in proopiomelanocortin neurons of the rat arcuate nucleus.

It has been previously demonstrated that the activity of proopiomelanocortin (POMC)-containing neurons in the rat arcuate nucleus is regulated by opiates, but the expression of opioid receptors in POMC neurons has never been reported. In the present study, we have applied a double-labeling in situ hybridization technique to investigate the occurrence of mu-opioid receptor mRNA on POMC neurons. We have found that 20+/-3% of arcuate POMC neurons express mu-opioid receptor mRNA and that the proportion of POMC neurons expressing mu-opioid receptor is higher in the caudal than in the rostral portion of the arcuate nucleus. Our data suggest that POMC neurons might be both auto-regulated by beta-endorphin, and regulated by enkephalins.

Animals↗

PubMed1999

Somatostatin binding sites in the white matter of the developing human brainstem: inverse relationship with the myelination process.

The ontogeny of somatostatin binding sites was studied in eight fiber tracts of the human lower brainstem in 18 fetuses and infants aged from 21 weeks postconceptional to 6 months postnatal, and in two adults. The study was performed by means of quantitative autoradiography using [125I-Tyr0,DTrp8]somatostatin-14 as a radioligand. For all structures examined, the highest densities of binding sites were detected in the younger stages and the density of sites decreased during development. These results reveal the existence of a close inverse relationship between the density of somatostatin receptors and the myelination process in the fiber tracts of the human brainstem.

Aged↗

PubMed1999

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.

Amino Acid Sequence↗

PubMed1999

GABA inhibits endozepine release from cultured rat astrocytes.

In the mammalian brain, the endogenous ligands for benzodiazepine receptors (also called endozepines) are predominantly synthesized by glial cells. It has recently been reported that rat astrocytes in primary culture release substantial amounts of endozepines. The aim of the present study was to investigate the possible involvement of GABA in the control of endozepine release. Exposure of cultured rat astrocytes to GABA (10(-7) to 10(-5) M) induced a dose-related inhibition of endozepine secretion. At higher doses (3 x 10(-5) to 10(-3) M), the effect of GABA gradually diminished. The inhibitory effect of GABA (10(-5) M) was mimicked by the GABA(B) receptor agonist baclofen (10(-5) M). In contrast, the GABA(A) receptor agonists 3APS and isoguvacine (10(-5) M each) did not modify endozepine release. The inhibition of endozepine secretion evoked by GABA and baclofen (10(-5) M each) was totally abrogated by the specific GABA(B) receptor antagonist phaclofen (10(-4) M). GABA and baclofen caused a significant inhibition of forskolin-evoked production of cAMP in astrocytes and this effect was abolished in the presence of phaclofen. In contrast, isoguvacine had no effect on cAMP production. Exposure of astrocytes to dbcAMP induced a time- and dose-dependent stimulation of endozepine release. These data indicate that GABA, acting through GABA(B) receptors negatively coupled to adenylyl cyclase, inhibits endozepine release from cultured rat astrocytes. The secretion of endozepines thus appears to be a valuable marker to monitor astrocyte activity.

Acyl Coenzyme A↗

PubMed1999

The stimulatory effect of endothelin-1 on frog adrenocortical cells is mediated through both the phospholipase C and the adenylyl cyclase transduction pathways.

We have previously shown that endothelin-1 (ET-1) stimulates corticosterone and aldosterone secretion by the frog adrenal gland through activation of ET(A) receptors. In the present study, we have investigated the transduction pathways involved in the corticotropic action of ET-1. Exposure of frog adrenal explants to ET-1 provoked a time- and dose-dependent increase in inositol phosphate production and a parallel decrease in membrane polyphosphoinositide content. Incubation of adrenal explants with ET-1 also induced a dose-related increase of cAMP formation. The selective ET(A) receptor antagonist BQ-485 totally abolished the stimulatory effects of ET-1 on both inositol phosphate and cAMP production. In contrast, the selective ET(B) receptor agonist IRL 1620 did not significantly modify polyphosphoinositide hydrolysis or cAMP formation. Administration of the phospholipase C inhibitor U-73122 or the protein kinase A inhibitor H-89 to perifused frog adrenal slices significantly reduced the stimulatory effect of ET-1 on corticosterone and aldosterone secretion. Concomitant administration of the two inhibitors almost completely suppressed the corticotropic effect of ET-1. Taken together, these data indicate that, in the frog adrenal gland, the stimulatory effect of ET-1 on corticosteroid secretion is mediated through activation of both the phospholipase C and the adenylyl cyclase transduction pathways.

Adenylyl Cyclases↗

PubMed1999

Immunohistochemical localization and biochemical characterization of two novel decapeptides derived from POMC-A in the trout hypothalamus.

Several vertebrate species which underwent duplication of their genome, such as trout, salmon and Xenopus, possess two proopiomelanocortin (POMC) genes. In the trout, one of the POMC molecules, called POMC-A, exhibits a unique C-terminal extension of 25 amino acids which has no equivalent in other POMCs characterized so far. This C-terminal peptide contains three pairs of basic residues, suggesting that it may be the source of novel regulatory peptides. The aim of the present study was to investigate the occurrence of these peptides in the brain of the trout Oncorhynchus mykiss by using specific antibodies raised against two epitopes derived from the C-terminal extension of POMC-A, i.e., EQWGREEGEE and YHFQ-NH2. Immunohistochemical labeling of brain sections revealed the presence of EQWGREEGEE- and YHFQ-NH2-immunoreactive cell bodies in the anterior part of the nucleus lateralis tuberis of the hypothalamus. Immunoreactive fibers were observed in the dorsal hypothalamus, the thalamus, the telencephalon, the optic tectum and the medulla oblongata. In contrast, no labeling was detected using antibodies against the non-amidated peptide YHFQG. Biochemical characterization was performed by combining high-performance liquid chromatography (HPLC) analysis with radioimmunoassay (RIA) quantification. Two peptides exhibiting the same retention time as synthetic EQWGREEGEE and ALGERKYHFQ-NH2 were resolved. However, no peptide co-eluting with YHFQ-NH2 or YHFQG could be detected. These results demonstrate that, in the trout brain, post-translational processing of POMC-A generates the two decapeptides EQWGREEGEE and ALGERKYHFQ-NH2. The wide distribution of immunoreactive fibers in the diencephalon, telencephalon, optic tectum and medulla oblongata suggests that these peptides may exert neurotransmitter and/or neuromodulator activities.

Animals↗

PubMed1999

The endozepine ODN stimulates [3H]thymidine incorporation in cultured rat astrocytes.

High concentrations of diazepam-binding inhibitor (DBI) mRNA have been detected in astrocytoma, suggesting that DBI-derived peptides may play a role in glial cell proliferation. In the present study, we have investigated the effect of a processing product of DBI, the octadecaneuropeptide ODN, on DNA synthesis in cultured rat astrocytes. At very low concentrations (10(-14) to 10(-11) M), ODN caused a dose-dependent increase of [3H]thymidine incorporation. At higher doses (10(-10) to 10(-5) M), the effect of ODN gradually declined. The central-type benzodiazepine receptor antagonist flumazenil (10(-6) M) completely suppressed the stimulatory action of ODN whereas the peripheral-type benzodiazepine receptor ligand, PK11195 (10(-6) M) had no effect. The ODN-induced stimulation of [3H]thymidine incorporation was mimicked by methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM). The GABAA receptor antagonist bicuculline (10(-4) M) suppressed the effect of both ODN and DMCM on DNA synthesis. Exposure of cultured astrocytes to the specific GABAA agonist 3APS (10(-10) to 10(-4) M) also induced a dose-related increase of [3H]thymidine incorporation. The present study indicates that ODN, acting through central-type benzodiazepine receptors associated with the GABAA receptor complex, stimulates DNA synthesis in rat glial cells. These data provide evidence for an autocrine role of endozepines in the control of glial cell proliferation.

Acyl Coenzyme A↗

PubMed1999

The proenkephalin A-processing product peptide E, which encompasses two enkephalin sequences, has a much lower opioid activity than beta-endorphin.

Peptide E is a 25-amino acid peptide derived from proenkephalin A that was originally isolated from the bovine adrenal medulla. Bovine peptide E (BPE), which possesses a Met- and a Leu-enkephalin sequence at its N- and C-terminus, respectively, has been described as a highly potent and selective mu-opioid receptor agonist. Paradoxically, the frog counterpart of peptide E (FPE), which exhibits only two amino acid substitutions (Met15-->Gln and Leu25-->Met) compared with BPE, was found to be totally devoid of antinociceptive activity. To decipher this apparent discrepancy, we have decided to compare the structural and pharmacological characteristics of FPE, BPE, and the chimeric peptide [Gln15]BPE (Q15BPE). In methanol, all three peptides exhibited virtually the same conformation, the central region of each peptide (residues 10-20) being involved in a regular helix. Intracerebroventricular administration of FPE, BPE, or Q15BPE, at doses up to 1000 ng per mouse, did not induce any analgesic effects, as evaluated by the hot plate and writhing tests, whereas, in the same tests, beta-endorphin at a dose of 100 ng provoked profound analgesia. Concomitant administration of FPE, BPE, or Q15BPE (100 ng) with the aminopeptidase-N inhibitor bestatin (50 microg) or the endopeptidase 24-11 inhibitor thiorphan (10 microg) did not produce analgesic responses. Antinociceptive effects were only observed when very high doses of FPE, BPE, and Q15BPE (10000 ng per mouse) were administered. These data clearly demonstrate that, contrary to what has been previously reported, peptide E is virtually devoid of opioid activity.

Amino Acid Sequence↗