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1.
J Neuroimmunol ; 76(1-2): 145-52, 1997 Jun.
Article in English | MEDLINE | ID: mdl-9184644

ABSTRACT

Long-term regulation of opioid binding was studied in the human neuroblastoma NMB and in the murine lymphoma R1.1 and R1.EGO cell lines. Binding was down-regulated following prolonged exposure to opioid agonists and up-regulated following exposure to antagonist. Down-regulation was inhibited by the metabolic blocker sodium-azide and by the protein kinase H-7. Up-regulation was blocked by the protein and mRNA synthesis blockers cycloheximide, alpha-amanitin and actinomycin D. A significant difference was found between the response of neuronal and immune cells to ethanol exposure: while opioid binding in neuroblastoma culture underwent a pronounced (75%) up-regulation, no effect of ethanol on opioid receptors in lymphoma cultures was detected. The described cell lines present an excellent experimental model to study long-term regulation of opioid receptors in the nervous and immune systems and to elucidate the biological effects of chronic use of opiates and alcohol.


Subject(s)
Lymphoma/chemistry , Neuroblastoma/chemistry , Receptors, Opioid/analysis , Diprenorphine/metabolism , Down-Regulation , Ethanol/pharmacology , Humans , Receptors, Opioid/drug effects , Tumor Cells, Cultured
2.
Brain Res ; 665(1): 94-100, 1994 Nov 28.
Article in English | MEDLINE | ID: mdl-7882024

ABSTRACT

The human neuroblastoma NMB cell line was found to contain the three types of opioid receptors (60% delta 25% kappa and 15% mu). The opioid receptors were negatively coupled to adenylyl-cyclase. Maximal reduction in cAMP content was achieved by selectively activating single receptor types, indicating the co-presence of the various opioid receptors in the same cells. The opioid receptors in NMB cells were up-regulated following prolonged exposure to the opioid antagonist naloxone and down-regulated following chronic treatment with the opioid agonist etorphine. Down-regulation was time-, dose- and temperature-dependent and was inhibited by colchicine and sodium azide. The NMB culture is presented as an excellent experimental model for studying the selective activation and regulation of the different opioid receptor types when they are co-expressed in the same neuron, as well as for studying interactions between the various opioid receptors.


Subject(s)
Adenylyl Cyclases/metabolism , Neurons/metabolism , Receptors, Opioid, delta/metabolism , Receptors, Opioid, kappa/metabolism , Receptors, Opioid, mu/metabolism , Amino Acid Sequence , Basal Metabolism , Cyclic AMP/metabolism , Down-Regulation/drug effects , Etorphine/pharmacology , Humans , Molecular Sequence Data , Neuroblastoma , Radioligand Assay , Tumor Cells, Cultured
3.
Mol Pharmacol ; 44(2): 461-7, 1993 Aug.
Article in English | MEDLINE | ID: mdl-8394999

ABSTRACT

Human neuroblastoma SK-N-SH cells, which contain both mu- and delta-opioid receptors, were grown under conditions that provided a mu:delta ratio of 1.5:1. Both receptors were down-regulated after 72 hr of exposure to 100 nM etorphine. Selective down-regulation was demonstrated using selective opioid agonists; the mu agonist Tyr-D-Ala2-Gly-(Me)Phe4-Gly-ol down-regulated mu- but not delta-opioid receptors, whereas prolonged exposure to the selective delta agonist D-Pen2,D-Pen5-enkephalin resulted in delta- but not mu-opioid receptor down-regulation. Morphine, which binds mu- as well as delta-opioid receptors, down-regulated both receptor subtypes. NG108-15 cells, which contain delta receptors exclusively, were also tested. NG108-15 cells did not exhibit delta-opioid receptor down-regulation when exposed to morphine. The discrepancy between the effect of chronic morphine treatment on delta receptors in SK-N-SH cells and in NG108-15 cells raised the question of whether the coexistence of mu receptors in the former allowed morphine to down-regulate delta receptors. The role of mu-opioid receptors in morphine-induced delta receptor down-regulation was studied by using the irreversible mu antagonist beta-funaltrexamine. Pretreatment of SK-N-SH cells with beta-funaltrexamine prevented down-regulation of delta receptors in response to chronic exposure to morphine but did not affect down-regulation of delta receptors in response to D-Pen2,D-Pen5-enkephalin. The experimental data indicate that morphine-induced delta-opioid receptor down-regulation is dependent on the presence of functional mu receptors in the same cell.


Subject(s)
Analgesics/pharmacology , Receptors, Opioid, delta/drug effects , Receptors, Opioid, mu/drug effects , 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer , Down-Regulation/drug effects , Enkephalin, Ala(2)-MePhe(4)-Gly(5)- , Enkephalin, D-Penicillamine (2,5)- , Enkephalins/pharmacology , Humans , Morphine/pharmacology , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Neuroblastoma/pathology , Pyrrolidines/pharmacology , Receptors, Opioid, delta/metabolism , Receptors, Opioid, mu/metabolism , Tumor Cells, Cultured
4.
Life Sci ; 52(19): PL205-10, 1993.
Article in English | MEDLINE | ID: mdl-8387137

ABSTRACT

Human neuroblastoma cells were tested for the presence of opioid receptors. [3H]Diprenorphine binds to NMB cell membranes with a KD value of 0.46 +/- 0.13 nM and Bmax of 534 +/- 22 fmol/mg protein. The presence of mu, delta, and kappa opioid receptors was tested by displacing [3H]diprenorphine specific binding by the selective agonists DAMGO, DPDPE, and U50,488H, respectively. Using this procedure, the data suggest that the NMB neuroblastoma cells express the three opioid receptor types with the abundance of delta receptors (about 60%) and minor, yet substantial populations of mu and kappa receptors (about 20% each).


Subject(s)
Neuroblastoma/ultrastructure , Receptors, Opioid/analysis , 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer , Analgesics/metabolism , Analgesics/pharmacology , Binding, Competitive , Diprenorphine/metabolism , Diprenorphine/pharmacology , Enkephalin, Ala(2)-MePhe(4)-Gly(5)- , Enkephalin, D-Penicillamine (2,5)- , Enkephalins/metabolism , Enkephalins/pharmacology , Humans , Kinetics , Neuroblastoma/metabolism , Pyrrolidines/metabolism , Pyrrolidines/pharmacology , Receptors, Opioid/metabolism , Receptors, Opioid, delta/analysis , Receptors, Opioid, delta/metabolism , Receptors, Opioid, kappa/analysis , Receptors, Opioid, kappa/metabolism , Receptors, Opioid, mu/analysis , Receptors, Opioid, mu/metabolism , Sensitivity and Specificity , Tritium , Tumor Cells, Cultured
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