dynorphins and adrenorphin

The ability of a number of opioid agonists and antagonists to affect nicotine-induced 45Ca2(+)-uptake into cultured bovine adrenal medullary cells has been investigated. High (10 microM) concentrations of the opioid agonist bremazocine produced a significant inhibition of nicotine-induced 45Ca2(+)-uptake throughout the 15 min time course examined. The opioid subtype-selectivity of this inhibition was investigated; mu and delta selective agonists produced only minor effects whereas the kappa selective agonist U50-488H and the endogenous opioid peptides dynorphin(1-13) and metorphamide almost abolished nicotine-induced 45Ca2(+)-uptake. The U50-488H inhibition was significant at 10 nM concentrations with an IC50 of approximately 1 microM. U50-488H inhibition could not be reversed or reduced by the opioid antagonists naxolone, diprenophine or Mr2266. Furthermore, Mr2266 and its optical isomer Mr2267 also produced marked inhibition of 45Ca2(+)-uptake. The inhibition was specific to nicotine-induced 45Ca2(+)-uptake in that a similar level of uptake evoked by potassium depolarization was unaffected by high concentrations of U50-488H. These data indicate that opioid inhibition of nicotine-induced 45Ca2(+)-uptake does not involve classical, stereospecific opioid receptors and suggests the involvement of a pharmacologically distinct opioid recognition site. It is speculated that this may be associated with the nicotine receptor-ionophore complex.

Topics: Adrenal Medulla; Animals; Benzomorphans; Calcium; Cattle; Cells, Cultured; Chromaffin System; Dynorphins; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Methionine; Enkephalins; Nicotine; Oligopeptides

The opioid peptides are potent inhibitors of gastric somatostatin-like immunoreactivity (SLI) secretion from the isolated perfused rat stomach. In addition, inhibition of SLI secretion induced by vagal stimulation is partially blocked by naloxone, indicating that endogenously released opioid peptides probably play a physiological role in the regulation of SLI release. The opioid peptides exert their effects by interacting with a number of different receptor types. In the present study, the effect of the selective delta-opioid receptor agonists [D-Pen2.5]enkephalin and [D-Pen2,L-Pen5]enkephalin and the mu-receptor agonist [D-Ala2, N-methyl (NMe)-Phe4,Gly5-ol]enkephalin on gastric inhibitory polypeptide (GIP)-stimulated SLI secretion from the isolated perfused rat stomach have been studied. Responses to the less selective delta-agonist [D-Ala2,D-Leu5]enkephalin, dynorphins 1-8, 1-13, and 1-17, and the extended enkephalin forms Met-enkephalin-Arg6-Phe7,Met- enkephalin-Arg6-Gly7-Leu8, and Met-enkephalin-Arg6-Arg7-Val8-NH2 (metorphamide), have also been investigated. [D-Ala2,NMe-Phe4,Gly5-ol]enkephalin induced a concentration-dependent inhibition of GIP-stimulated SLI secretion, with 50% of maximal inhibition at 10 nM. Neither [D-Pen2.5]enkephalin nor [D-Pen2,L-Pen6]enkephalin (10 nM to 1 microM) had any effect on SLI release, and [D-Ala2,D-Leu5] enkephalin inhibited SLI release only at high concentrations. Met-enkephalin-Arg6-Phe7 and metorphamide both inhibited SLI release, whereas Met-enkephalin-Arg6-Gly7-Leu8 and the dynorphins had little or no effect. In conclusion, the strong inhibition of SLI secretion produced by [D-Ala2,NMe-Phe4,Gly5-ol] enkephalin and lack of major effect of [D-Pen2.5]-enkephalin, [D-Pen2,L-Pen5]enkephalin, and the dynorphins indicate that opioid peptide-induced inhibition was mediated by interaction with mu-receptors and that neither delta or kappa-receptors play a significant role.

Topics: Animals; Dynorphins; Enkephalin, Methionine; Enkephalins; Gastric Inhibitory Polypeptide; Gastric Mucosa; In Vitro Techniques; Kinetics; Male; Rats; Rats, Inbred Strains; Receptors, Opioid; Somatostatin

It has been previously reported that both the cysteinyl-endo-oligopeptidase A and the metalloendopeptidase EC 3.4.24.15 are able to generate enkephalin from a number of enkephalin-containing peptides, including dynorphin A1-8. The present study shows that only endo-oligopeptidase A is able to generate [Leu5]enkephalin and [Met5]enkephalin from dynorphin A1-8 and from metorphamide respectively. It is also shown that endo-oligopeptidase A neither hydrolyses the specific EC 3.4.24.15 substrate alpha-N-benzoyl-Gly-Ala-Ala-Phe p-aminobenzoate, nor is inhibited by the specific EC 3.4.24.15 inhibitor N-[1(RS)-carboxy-2-phenylethyl]-alpha-Ala-Ala-Phe p-aminobenzoate.

Topics: Chemical Precipitation; Cysteine Endopeptidases; Cysteine Proteinase Inhibitors; Dynorphins; Enkephalin, Methionine; Enkephalins; Metalloendopeptidases; Peptide Fragments

Opioid compounds have been assessed for their ability to modify desensitization of nicotine-induced catecholamine secretion from cultured, bovine, adrenal chromaffin cells. Dynorphin-1-13 and metorphamide produced protection against desensitization of the nicotinic response at concentrations between 1 and 20 microM while etorphine and morphine only produced this effect at 100 microM. The opioid antagonists, naloxone and diprenorphine, at 100 microM mimicked the weak ability of the opioid agonists to protect against nicotinic desensitization. All opioid compounds tested were considerably more potent at inhibiting nicotine-induced catecholamine secretion from the cells than at protecting against desensitization of this response. It is concluded that adrenal opioid peptides probably do not act on adrenal opioid binding sites characterised from ligand binding studies to prevent the nicotinic response from desensitizing. They are unlikely, therefore, to be involved in such a mechanism to maintain catecholamine secretion during stress.

Topics: Adrenal Medulla; Animals; Calcium; Cattle; Cells, Cultured; Chromaffin System; Diprenorphine; Drug Tolerance; Dynorphins; Endorphins; Enkephalin, Methionine; Epinephrine; Etorphine; Morphine; Naloxone; Nicotine; Norepinephrine; Peptide Fragments; Potassium

Adrenorphin is the first C-terminally amidated form of opioid peptides isolated from human pheochromocytoma tumor and is considered to be generated out of proenkephalin A by unique processing. By the highly specific and sensitive radioimmunoassay (RIA) procedure utilizing the antiserum against adrenorphin, combined with high performance liquid chromatography (HPLC), immunoreactive adrenorphin in rat brain was verified to be identical with its authentic peptide. It has been revealed that adrenorphin immunoreactivity distributes widely in rat brain but in the unique pattern distinct from those of other endogenous opioid peptides. Note that immunoreactive adrenorphin was most concentrated in the olfactory bulb, and appreciably in the hypothalamus and striatum. Furthermore, immunohistochemical study has revealed that adrenorphin-immunoreactive structures in hypothalamic region of rat were localized in the neurones of the arcuate nucleus. In addition, adrenorphin-immunoreactive fibre plexus was found in the various regions of the hypothalamus, such as median eminence, periventricular zone and paraventricular nucleus. These indicate that adrenorphin may have a unique physiological function.

Topics: Animals; Brain Chemistry; Chromatography, High Pressure Liquid; Dynorphins; Endorphins; Enkephalin, Methionine; Fluorescent Antibody Technique; Hypothalamus; Peptide Fragments; Protein Precursors; Radioimmunoassay; Rats

Adrenorphin is the first C-terminally amidated form of opioid peptide isolated from human pheochromocytoma tumor and is considered to be generated out of proenkephalin A by unique processing. We have developed a highly specific and sensitive radioimmunoassay for adrenorphin as well as for PH- 8P , whose structure and processing are similar to adrenorphin . Prior to the measurement of both peptides in rat brain, immunoreactive adrenorphin and PH- 8P were verified to be identical with their individual authentic peptides by high performance liquid chromatography. Here we have determined the distribution of adrenorphin in rat brain by radioimmunoassay, and compared it with that of PH- 8P . The regional distribution of adrenorphin was found to be quite different from that of other endogenous opioid peptides including PH- 8P . The highest concentration of adrenorphin was found in the olfactory bulb. These results suggest that adrenorphin is generated by a specific processing mechanism and may have a unique physiological function distinct from that of known opioid peptides. In addition, we identified adrenorphin also in human and bovine adrenal medullas.

Topics: Animals; Antibody Specificity; Brain Chemistry; Chromatography, High Pressure Liquid; Dynorphins; Endorphins; Enkephalin, Methionine; Male; Peptide Fragments; Radioimmunoassay; Rats; Rats, Inbred Strains