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Ni-coupled receptors in cultured neural hybrid cells: cell specificity for dibutyryl cyclic AMP-induced down-regulation but not morphological differentiation.
Mol Pharmacol. 1986 Dec; 30(6):526-36.MP

Abstract

Opiate, muscarinic, and alpha 2-adrenergic receptors and the Ni-coupled response of adenylate cyclase (AC) inhibition were examined in neuroblastoma X glioma NG108-15 (108 CC15) and neuroblastoma X Chinese hamster brain NCB-20 clonal hybrid cells, induced to differentiate with 1.0 mM dibutyryl cAMP (dBcAMP). Scatchard analysis of binding of the opiate agonist 3H-(D-Ala2,D-Leu5)enkephalin (DADLE) and the antagonist [3H] diprenorphine to dBcAMP-treated NCB-20 cell membranes indicated an 80% reduction in opiate receptor density relative to untreated cells (Bmax = 47 +/- 11 fmol/mg of protein versus 220 +/- 48 fmol/mg of protein), with no change in ligand affinities. Binding of the muscarinic cholinergic antagonist [3H]quinuclidinyl benzilate and the alpha 2-adrenergic agonist [3H]-p-aminoclonidine to dBcAMP-treated NCB-20 membranes was also reduced by 50% and 28%, respectively. In contrast, treatment of NG108-15 cells with dBcAMP did not down-regulate opiate, muscarinic, or alpha 2-adrenergic receptor sites. Opiate and alpha 2-adrenergic receptor sites were not down-regulated in the N18TG2 neuroblastoma clone, the common parent of both the hybrid cells, and the apparent source of these receptors. The C6BU-1 parent of the NG108-15 hybrid showed poor specific binding of all ligands examined. dBcAMP was very potent in inducing opiate receptor site down-regulation of NCB-20 cells, with an ED50 after 4 days treatment of 8 microM. The time course of loss of [3H]DADLE and [3H]quinuclidinyl benzilate specific binding was similar, and maximum down-regulation was achieved after 2 days. In contrast, neither higher concentrations of dBcAMP (5.0 mM) nor longer treatment times (7 days) resulted in down-regulation of receptor sites on NG108-15 cells. Coupling of opiate receptors to AC was also selectively altered in differentiated NCB-20 cells. Prostaglandin E1-stimulated AC was maximally inhibited by 1 microM DADLE in membranes from undifferentiated cells to different degrees (30% in NCB-20 and 54% in NG108-15). dBcAMP treatment had no effect on opiate inhibition of AC in NG108-15 cells but reduced by 50% the maximum opiate inhibition of AC in NCB-20 cells. These data indicate that the signal for receptor down-regulation which was triggered by dBcAMP in the NCB-20 cell comes uniquely from the Chinese hamster brain cell NCB-20 parent. The differences between NCB-20 and NG108-15 cells in the regulation of Ni-coupled receptors provides an example of dBcAMP-induced heterologous down-regulation with unique cell specificity, which is unrelated to the morphological differentiation process triggered by dBcAMP, which is common to both cells.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

3023808

Citation

Noronha-Blob, L, et al. "Ni-coupled Receptors in Cultured Neural Hybrid Cells: Cell Specificity for Dibutyryl Cyclic AMP-induced Down-regulation but Not Morphological Differentiation." Molecular Pharmacology, vol. 30, no. 6, 1986, pp. 526-36.
Noronha-Blob L, Lowe VC, Kinnier WJ, et al. Ni-coupled receptors in cultured neural hybrid cells: cell specificity for dibutyryl cyclic AMP-induced down-regulation but not morphological differentiation. Mol Pharmacol. 1986;30(6):526-36.
Noronha-Blob, L., Lowe, V. C., Kinnier, W. J., & U'Prichard, D. C. (1986). Ni-coupled receptors in cultured neural hybrid cells: cell specificity for dibutyryl cyclic AMP-induced down-regulation but not morphological differentiation. Molecular Pharmacology, 30(6), 526-36.
Noronha-Blob L, et al. Ni-coupled Receptors in Cultured Neural Hybrid Cells: Cell Specificity for Dibutyryl Cyclic AMP-induced Down-regulation but Not Morphological Differentiation. Mol Pharmacol. 1986;30(6):526-36. PubMed PMID: 3023808.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Ni-coupled receptors in cultured neural hybrid cells: cell specificity for dibutyryl cyclic AMP-induced down-regulation but not morphological differentiation. AU - Noronha-Blob,L, AU - Lowe,V C, AU - Kinnier,W J, AU - U'Prichard,D C, PY - 1986/12/1/pubmed PY - 1986/12/1/medline PY - 1986/12/1/entrez SP - 526 EP - 36 JF - Molecular pharmacology JO - Mol Pharmacol VL - 30 IS - 6 N2 - Opiate, muscarinic, and alpha 2-adrenergic receptors and the Ni-coupled response of adenylate cyclase (AC) inhibition were examined in neuroblastoma X glioma NG108-15 (108 CC15) and neuroblastoma X Chinese hamster brain NCB-20 clonal hybrid cells, induced to differentiate with 1.0 mM dibutyryl cAMP (dBcAMP). Scatchard analysis of binding of the opiate agonist 3H-(D-Ala2,D-Leu5)enkephalin (DADLE) and the antagonist [3H] diprenorphine to dBcAMP-treated NCB-20 cell membranes indicated an 80% reduction in opiate receptor density relative to untreated cells (Bmax = 47 +/- 11 fmol/mg of protein versus 220 +/- 48 fmol/mg of protein), with no change in ligand affinities. Binding of the muscarinic cholinergic antagonist [3H]quinuclidinyl benzilate and the alpha 2-adrenergic agonist [3H]-p-aminoclonidine to dBcAMP-treated NCB-20 membranes was also reduced by 50% and 28%, respectively. In contrast, treatment of NG108-15 cells with dBcAMP did not down-regulate opiate, muscarinic, or alpha 2-adrenergic receptor sites. Opiate and alpha 2-adrenergic receptor sites were not down-regulated in the N18TG2 neuroblastoma clone, the common parent of both the hybrid cells, and the apparent source of these receptors. The C6BU-1 parent of the NG108-15 hybrid showed poor specific binding of all ligands examined. dBcAMP was very potent in inducing opiate receptor site down-regulation of NCB-20 cells, with an ED50 after 4 days treatment of 8 microM. The time course of loss of [3H]DADLE and [3H]quinuclidinyl benzilate specific binding was similar, and maximum down-regulation was achieved after 2 days. In contrast, neither higher concentrations of dBcAMP (5.0 mM) nor longer treatment times (7 days) resulted in down-regulation of receptor sites on NG108-15 cells. Coupling of opiate receptors to AC was also selectively altered in differentiated NCB-20 cells. Prostaglandin E1-stimulated AC was maximally inhibited by 1 microM DADLE in membranes from undifferentiated cells to different degrees (30% in NCB-20 and 54% in NG108-15). dBcAMP treatment had no effect on opiate inhibition of AC in NG108-15 cells but reduced by 50% the maximum opiate inhibition of AC in NCB-20 cells. These data indicate that the signal for receptor down-regulation which was triggered by dBcAMP in the NCB-20 cell comes uniquely from the Chinese hamster brain cell NCB-20 parent. The differences between NCB-20 and NG108-15 cells in the regulation of Ni-coupled receptors provides an example of dBcAMP-induced heterologous down-regulation with unique cell specificity, which is unrelated to the morphological differentiation process triggered by dBcAMP, which is common to both cells. SN - 0026-895X UR - https://www.unboundmedicine.com/medline/citation/3023808/Ni_coupled_receptors_in_cultured_neural_hybrid_cells:_cell_specificity_for_dibutyryl_cyclic_AMP_induced_down_regulation_but_not_morphological_differentiation_ L2 - http://molpharm.aspetjournals.org/cgi/pmidlookup?view=long&pmid=3023808 DB - PRIME DP - Unbound Medicine ER -