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E Costa

Publications and source records attributed to E Costa.

At least 361 records · Page 20Linked to original sources
PubMed1983

Substance P-induced release of Met5-enkephalin from striatal and periaqueductal gray slices.

Substance P(SP), the heptapeptide SP and the stable analogue (p-Glu5-MePhe8-MeGly9) SP (DiMe-C7) induce a Ca2+-dependent release of Met5-enkephalin (MET) from slices of periaqueductal gray matter (PAG) and striatum of rats. The MET release from striatal slices is greater than that from PAG slices because of the higher MET content of striatum. Intraventricular injection of SP and of the two related peptides induce analgesia in the rat, and their analgesic potency is in line with their capacity to release MET. Other neuropeptides which possess antinociceptive activity such as bombesin, neurotensin, vasopressin and somatostatin fail to release MET from PAG slices.

Animals↗
PubMed1983

Met5-enkephalin-arg6-phe7 content of human and rabbit plasma.

Using an adsorption technique combined with high pressure liquid chromatography and a specific radioimmunoassay, the met5-enkephalin-arg6-phe7 (YGGFMRF) content was measured in human and rabbit plasma. This heptapeptide content was 0.16 +/- 0.03, 0.20 +/- 0.05 pmol/10 ml plasma, for human and rabbit plasma, respectively. The degradation of YGGFMRF injected intravenously (rabbit) or that of the opioid heptapeptide added to rabbit plasma is rapid. The biological half life (T 1/2) of 125I-YGGFMRF in rabbit plasma was about 45 seconds (in vivo) and 1 minute and 8 seconds (in vitro). The metabolic clearance rate of YGGFMRF is slower than that of met5-enkephalin. The YGGFMRF content of rabbit plasma increased following inhibition of dipeptidyl peptidase activity by an intravenous injection of captopril. The presence of met5-enkephalin-arg6-phe7 into circulation indicates that it may have some physiological role, however it is unknown whether circulating YGGFMRF originates from adrenal medulla, intermediate lobe of pituitary or some other site.

Adult↗
PubMed1983

Met5-enkephalin-arg6-phe7 and its receptor in lung.

The presence of met5-enkephalin-arg6-phe7 (YGGFMRF) and opiate receptors in rat, guinea pig and human lung was investigated with specific and sensitive radio immuno- and radio-receptor assays. 1) High and low molecular weight YGGFMRF-like immunoreactivity were detected in lung extracts using Bio-Gel P-2 column chromatography followed by radioimmunoassay. Using HPLC, we determined that the low molecular weight YGGFMRF-like immunoreactivity is authentic YGGFMRF. 2) The contents of YGGFMRF were 0.68 +/- 0.08, 0.76 +/- 0.12 and 0.63 pmol/mg protein in lung of rat, guinea pig and human, respectively. In the lung of these three species, the content of YGGFMRF is much greater than that of met5-enkephalin. 3) 47 mM KCl released YGGFMRF from rat lung slices in a Ca++ dependent manner. 4) Rat lung membranes were shown to bind [3H]-etorphine in a saturable manner. There are two populations of binding sites with a Kd = 0.6 and 7.1 nM and a Bmax = 7.8 and 28.5 fmol/mg protein, respectively. This binding could be displaced by YGGFMRF with high affinity, the other endogenous opioid peptides were poor displacers. From these results, we infer that YGGFMRF might be a putative neurotransmitter or neuromodulator, its role in the regulation of lung function can now be investigated.

Animals↗
PubMed1983

(-)-Deprenyl a selective MAO "B' inhibitor, increases [3H]imipramine binding and decreases beta-adrenergic receptor function.

In rats, a selective inhibition for 3 weeks of monoamineoxydase (MAO) type B elicited by daily doses of pargyline (2.5 mumol/kg) or (-)-deprenyl (1 mumol/kg) attenuated the NE dependent stimulation of cortical adenylate cyclase and reduced the number of brain recognition sites for beta-adrenergic receptor ligands. Similar actions were not elicited by a comparable dose regimen of (+)-amphetamine. Hence the inhibition of MAO B mimicks responses that are typically elicited by antidepressants. The molecular nature of the mechanisms involved cannot be understood, however, these mechanisms may not be identical for pargyline and (-)-deprenyl because this drug but not pargyline increased the number of [3H]imipramine recognition sites. Even high daily doses of pargyline (100 mumol/kg, for 3 weeks) failed to change [3H]imipramine binding though they still down regulated beta-adrenergic recognition sites, the NE stimulation of adenylate cyclase and the Bmax of [3H]mianserin and [3H]spiroperidol binding.

Animals↗
PubMed1983

The transsynaptic regulation of the septal-hippocampal cholinergic neurons.

There is not yet a complete understanding of the functional interactions among various septal nuclei which regulate hippocampal function. Nevertheless, much has been learned histologically and biochemically about the major connections of the distinct areas of the septal complex and the chemical character of some of these pathways. The cholinergic septal-hippocampal pathway serves as a well defined link between these two important structures of the limbic system. Acetylcholine turnover rates in the hippocampus have been shown to increase or decrease proportionally to the activity of the cholinergic neurons originating in the septum. Moreover, these turnover rates have been shown to be modulated by intraseptal injections of agonists or antagonists of various neurotransmitters or neuromodulators which are stored in various cell groups located in the septum. By coupling this biochemical approach with techniques to study the receptor organization, greater detail concerning the transmitter and cotransmitter interactions among the various neuromodulators can be obtained.

Acetylcholine↗
PubMed1983

A high molecular weight form of met5-enkephalin-arg6-gly7-leu8 in rat brain and bovine adrenal chromaffin granules.

An antiserum against met5-enkephalin-arg6-gly7-leu8 (MAGL) has been used to identify various molecular weight forms of this immunoreactive material in brain and adrenal tissue. The predominant immunoreactive species in acid extracts of chromaffin granules eluted on Biogel P-30 as an approximately equal to 8,000 dalton peptide. Acid extracts prepared from rat brain medulla-pons also contained 8,000 dalton immunoreactive material which exhibited an identical retention time on HPLC to the corresponding immunoreactive material prepared from chromaffin granules.

Adrenal Medulla↗
PubMed1983

The effect of peptidase inhibitors on the release of Met5-Enk-Arg6-Phe7 (YGGFMRF) and Met5-enkephalin (YGGFM) from spinal cord induced by substance P in vivo.

Using a preparation for the perfusion of the subarachnoidal spaces of the spinal cord of rats it was found that substance P can stimulate the release of YGGFMRF and YGGFM. We have studied the effect of several peptidase inhibitors (captopril, bestatin, thiorphan) on the recovery of YGGFMRF and YGGFM released from spinal cord by substance P. The recovery of released YGGFMRF was increased by adding captopril to the perfusion medium. A combination of captopril and bestatin in the perfusion medium further increases this YGGFMRF recovery. Intrathecal injection of captopril and bestatin also potentiated the analgesic effect of YGGFMRF and electroacupuncture. These results suggest that substance P may act as a "releaser" of enkephalins in spinal cord and that the dipeptidyl carboxypeptidase and aminopeptidase may be important in the degradation of YGGFMRF in vivo.

Animals↗
PubMed1983

Substance P stimulates the release of Met5-enkephalin-Arg6-Phe7 and Met5-enkephalin from rat spinal cord.

Substance P (10(-6) M) injected in the fluid perfusing the subarachnoidal spaces of the spinal cord can release met5-enkephalin (YGGFM) and met5-enkephalin-Arg6-Phe7 (YGGFMRF) from spinal cord. This effect is Ca2+ dependent and can be blocked by the substance P antagonist, D-pro2, -D Trp7,9-substance P. The release of YGGFM and YGGFMRF was not observed when substance P was replaced by substance P1-9. These results suggest that substance P receptors are involved in triggering the release of enkephalins.

Animals↗
PubMed1983

In vivo modulation of brain dopamine recognition sites: a possible model for emission computed tomography studies.

The content of authentic 3H-spiroperidol and of its metabolites was measured in brain regions of rat, guinea pig and mouse receiving tracer doses of 3H-spiroperidol intravenously (0.2 to 0.5 micrograms/kg). The time course of the 3H-spiroperidol content of various brain regions shows that a steady state was maintained between 2 and 6 hrs; the lowest 3H-spiroperidol content was attained in cerebellum where the value approached that of blood plasma. Since the cerebellum contains an insignificant number of dopamine receptors but many serotonin receptors and other sites that bind 3H-spiroperidol, the 3H-spiroperidol contained in cerebellum was considered background binding. In the striatum and olfactory tubercle of rats receiving two daily doses for 3 weeks of haloperidol or amphetamine the amount of 3H-spiroperidol that binds in vivo is decreased or increased, respectively. If the kinetic characteristics of in vivo binding of 3H-spiroperidol observed in the rat, guinea pig and mouse can be replicated in man using spiroperidol containing a gamma- or a position-emitting label, one might have a probe to study dopamine receptors in vivo with emission computed tomography scanning.

Animals↗
PubMed1983

GABAergic synapses. Supramolecular organization and biochemical regulation.

Extraneurally released gamma-aminobutyric acid (GABA) interacts with specific recognition sites associated with proteins located in postsynaptic neuronal membranes that function as chloride (Cl-)ionophores. As a result of the interaction between GABA and the recognition sites, Cl- ionophores are opened causing an influx or an efflux of Cl-, depending on the values of the Cl- equilibrium potential and of the membrane potential. Hyperpolarization or depolarization will result from inward or outward Cl- fluxes, respectively. Independently of the change in conductivity elicited by GABA, this amino acid transmitter will reduce the effectiveness of the sodium ion (Na+) excitatory potential. In attempts to elucidate the molecular mechanism, whereby benzodiazepines facilitate the action of GABA on membrane conductance without changing the activity of Cl- or other ionophore, a basic protein (GABA-modulin, GM) has been isolated from rat brain which is similar in structure to the small molecular weight myelin basic protein, found in rodent brain. While GABA-modulin is located in synaptosomes, the small molecular weight myelin basic protein is located in the myelin fraction: more important, GABA-modulin inhibited the high affinity binding of GABA to crude synaptic membranes while the basic myelin protein did not. Also, amino acid composition and molecular weight differentiate the two proteins. The GABA-modulin can be phosphorylated with different stoichiometry by cyclic AMP-dependent protein kinase (4 mol PO4(-3)) or Ca2+-dependent protein kinase (1 mol PO4(-3)). Only cyclic AMP-dependent phosphorylation inhibited the action of GABA-modulin on GABA binding.

Animals↗
PubMed1983

On a brain polypeptide functioning as a putative effector for the recognition sites of benzodiazepine and beta-carboline derivatives.

Stimulation of benzodiazepine recognition sites by various ligands can elicit opposite types of responses such as proconvulsant and anticonvulsant or proconflict and anticonflict actions. The study of the pharmacological profile of various ligands makes it possible to distinguish three classes of compounds: (1) those that elicit anticonflict responses in the Vogel test, inhibit the convulsions due to an impairment of GABAergic transmission and increase the Bmax of the high affinity recognition site for GABA; (2) those that displace benzodiazepines from specific binding sites, facilitate convulsions due to impairment of GABAergic mechanisms, elicit proconflict responses in Vogel's test and inhibit the facilitation by benzodiazepines of GABA binding and (3) those that displace benzodiazepines and beta-carboline-derivatives from specific binding sites, antagonize the anticonflict, the proconflict, the anticonvulsant and the proconvulsant actions of the two preceding groups of substances and in very large doses elicit a small proconvulsant action. Examples of the latter are an imidazobenzodiazepine (RO 15-1788) and a pyrazolquinolinone derivative (CGS 8216). The nomenclature for these three classes of drugs should be kept flexible until the action of the endogenous ligand that functions as the physiological effector of the benzodiazepine/beta-carboline recognition site is known. A putative ligand for this site (DBI = diazepam binding inhibitor) has been isolated and purified to homogeneity. It includes 104 amino acid residues, the sequence of the last 45 amino acids has been determined. This compound elicits a proconflict action, displaces beta-carboline derivatives more than anxiolytic benzodiazepines, but its high molecular weight and relative low affinity for the binding sites in brain might suggest that it is a precursor, rather than the putative effector for benzodiazepine and/or beta-carboline recognition sites.

Animals↗
PubMed1983

Daily bupropion injections for 3 weeks attenuate the NE stimulation of adenylate cyclase and the number of beta-adrenergic recognition sites in rat frontal cortex.

In rats receiving daily doses (50 mg/kg i.p. twice daily) of bupropion HCI (WellbutrinR) repeated for 21 days the Bmax of the beta-adrenergic receptor recognition sites located in the frontal cortex is reduced. This decrease is not associated with a decrease of the apparent affinity of these recognition sites. However the Vmax of the cAMP (cyclic AMP) generating system stimulated by NE is reduced suggesting that similarly to other antidepressants bupropion down regulates beta-adrenergic receptors located in the frontal cortex. Bupropion neither inhibits MAO (monoamine oxidase) nor releases biogenic amines but only weakly inhibits monoamine uptake in vitro.

Adenylyl Cyclases↗
PubMed1983

On the mode of action of imipramine: relationship between serotonergic axon terminal function and down-regulation of beta-adrenergic receptors.

Recognition sites for [3H]imipramine and [3H]mianserin are located in different structures and regulate different neuronal functions. Recognition sites for [3H]imipramine are located on serotonergic terminals, are part of the supramolecular organization of the uptake mechanisms and can be down-regulated by prolonged administration of the drug. When the number of recognition sites for imipramine is down-regulated, uptake of 5-hydroxytryptamine (5HT) in rat brain hippocampal slices is increased. The presence of the binding sites for imipramine in 5HT terminals is essential to mediate the down-regulation of recognition sites for norepinephrine (NE) and NE-mediated stimulation of adenylate cyclase. Mianserin binds on a site that is modulated by 5HT, the number of its binding sites is not down-regulated by repeated treatment and, like imipramine, decreases the NE-dependent cyclase but not the number of beta-adrenergic receptor recognition sites. Repeated treatment with imipramine and mianserin down-regulated the number of 5HT2 recognition sites. Several lines of evidence indicate that binding site for mianserin is related but not identical to the 5HT2 receptor binding site.

Animals↗
PubMed1983

Differences in the regulatory adaptation of the 5HT recognition sites labelled by 3H-mianserin or 3H-ketanserin.

In crude synaptic membranes prepared from rat brain the sites occupied by 3H-spiroperidol that are displaced by microM concentrations of serotonin (5HT) have been termed 5HT2 receptors (Peroutka and Snyder, 1980). Since the 3H-spiroperidol displaced by 5HT is also displaced very effectively (IC50 in the nM range) by ketanserin and mianserin it was suggested that spiroperidol, mianserin and ketanserin are labelling 5HT2 receptors. Data are presented showing that the 3H-ketanserin and 3H-mianserin bound to crude synaptic membrane in the presence of a H1 receptor blocker are not labelling the same recognition site. Hence from this standpoint the recognition site marked by 3H-mianserin and 3H-ketanserin is not identical. The possibility that allosteric effects are operative in some of these ligand displacements should be entertained.

Animals↗
PubMed1983

Isolation, characterization, and purification to homogeneity of an endogenous polypeptide with agonistic action on benzodiazepine receptors.

A brain polypeptide termed diazepam-binding inhibitor (DBI) and thought to be chemically and functionally related to the endogenous effector of the benzodiazepine recognition site was purified to homogeneity. This peptide gives a single band of protein on NaDodSO4 and acidic urea gel electrophoresis. A single UV-absorbing peak was obtained by HPLC using three different columns and solvent systems. DBI has a molecular mass of approximately equal to 11,000 daltons. Carboxyl-terminus analysis shows that tyrosine is the only residue while the amino-terminus was blocked. Cyanogen bromide treatment of DBI yields three polypeptide fragments, and the sequences of two of them have been determined for a total of 45 amino acids. DBI is a competitive inhibitor for the binding of [3H]diazepam, [3H]flunitrazepam, beta-[3H]carboline propyl esters, and 3H-labeled Ro 15-1788. The Ki for [3H]-diazepam and beta-[3H]carboline binding were 4 and 1 microM, respectively. Doses of DBI that inhibited [3H]diazepam binding by greater than 50% fail to change [3H]etorphine, gamma-amino[3H]butyric acid, [3H]-quinuclidinyl benzilate, [3H]dihydroalprenolol, [3H]adenosine, and [3H]imipramine binding tested at their respective Kd values. DBI injected intraventricularly at doses of 5-10 nmol completely reversed the anticonflict action of diazepam on unpunished drinking and, similar to the anxiety-inducing beta-carboline derivative FG 7142 (beta-carboline-3-carboxylic acid methyl ester), facilitated the shock-induced suppression of drinking by lowering the threshold for this response.

Amino Acids↗
PubMed1983

Increase of proenkephalin mRNA and enkephalin content of rat striatum after daily injection of haloperidol for 2 to 3 weeks.

Proenkephalin mRNA has been detected in striatum, hypothalamus, cortex, cerebellum, hippocampus, midbrain, and brain stem of rat by RNA ("Northern") blot analysis using a 918-base-pair DNA hybridization probe complementary to proenkephalin mRNA [Comb, M., Seeburg, P. H., Adelman, J., Eiden, L. & Herbert, E. (1982) Nature (London) 295, 663-666]. The size of the mRNA species in all brain regions is approximately 1,400 bases, and it was found to be comparable with that of bovine adrenal medulla and human pheochromocytoma. The data were quantified by densitometric scanning of the autoradiograms: the area under the peak is proportional to the amount of standard proenkephalin mRNA from bovine adrenal medulla. The relative content of proenkephalin mRNA in the various brain regions correlates generally with the content of [Met5]enkephalin-like immunoactivity of these regions. Rats receiving daily intraperitoneal injection of haloperidol (1 or 2 mg/kg) show a fourfold increase of proenkephalin mRNA content in striatum but not other brain regions. In agreement with previous reports, [Met5]enkephalin-like immunoactivity increased twofold in striatum.

Animals↗