ABSTRACT
Receptor binding studies were performed to characterize the properties of subtypes of kappa opioid receptors in membrane preparations of human cerebral cortex. [3H]U69,593 ([3H]U69), a selective kappa 1-agonist, and [3H]diprenorphine ([3H]DIP), a non-selective opioid antagonist, in the presence of 1 microM each of DAMGO, DPDPE and U-69 to block mu-, delta-, and kappa 1-sites, labeled single population of binding sites, respectively. [3H]U-69 binding sites (KD = 3.8 +/- 0.2 nM, Bmax = 6.3 +/- 0.2 fmol/mg protein) had a binding profile that correspond to kappa 1-receptor. That is, dynorphin A (1-13) (Dyn A), bremazocine (BZC), U50,488H (U50), (-)ethylketocyclazocine (EKC) and nor-binaltorphimine (nor-BNI) bound to this site with high affinities. [3H]DIP labeled binding sites (Kd = 7.3 +/- 0.2 nM, Bmax = 102 +/- 9 fmol/mg protein) that were not sensitive to U-50, but to BZC, EKC and nor-BNI. These results indicate that kappa 1 and Kappa 2 opioid receptors exist in human cerebral cortex with different ligand binding profiles.
Subject(s)
Analgesics/pharmacology , Benzeneacetamides , Cerebral Cortex/drug effects , Pyrrolidines/pharmacology , Receptors, Opioid, kappa/drug effects , Cell Membrane/drug effects , Cell Membrane/metabolism , Cerebral Cortex/metabolism , Diprenorphine/pharmacology , Humans , In Vitro Techniques , Protein Binding , Radioligand Assay , Receptors, Opioid, delta/antagonists & inhibitors , Receptors, Opioid, kappa/agonists , Receptors, Opioid, kappa/antagonists & inhibitors , Receptors, Opioid, mu/antagonists & inhibitors , TritiumABSTRACT
1. Ischaemia was induced by 5 min of deprivation of glucose and an additional 5 min of deprivation of glucose and oxygen from Mg(2+)-free artificial cerebrospinal fluid in vitro. 2. During the ischaemic period, 11 +/- 1.5% of the total [3H]-dopamine ([3H]-DA) was released into the incubation medium. 3. Ischaemia-evoked release of [3H]-DA from striatal slices was attenuated by tetrodotoxin (TTX), MgSO4, dizocilpine, ketamine, 6,7-dinitroquinoxaline-2,3-dione (DNQX) or carbetapentane. 4. Release of [3H]-DA was attenuated by verapamil, omega-conotoxin GVIA and dantrolene. 5. Nomifensin inhibited the ischaemia-induced release of [3H]-DA. 6. Omission of Ca(2+) from incubation media potentiated ischaemia-evoked [3H]-DA release. The inhibitory effect of nomifensin was potentiated in Ca(2+)-free incubation media. 7. These results suggest that ischaemia induces release of [3H]-DA by dual mechanisms; one is Ca(2+)-dependent exocytosis and the other is reversal of transporter.
