title: Phosphorylation- and voltage-dependent inhibition of neuronal calcium currents by activation of human D2(short) dopamine receptors.
abstract: 1. Activation of human D2(s) dopamine receptors with quinpirole (10 nM) inhibits omega-conotoxin GVIa-sensitive, high-threshold calcium currents when expressed in differentiated NG108-15 cells (55% inhibition at +10 mV). This inhibition was made irreversible following intracellular dialysis with the non-hydrolysable guanosine triphosphate analogue GTP-gamma-S (100 microM), and was prevented by pretreatment with pertussis toxin (1 microgram ml-1 for 24 h). 2. Stimulation of protein kinase C with the diacylglycerol analogue, 1-oleoyl-2-acetyl-sn-glycerol (100 microM), also attenuated the inhibition of the sustained calcium current but did not affect the receptor-mediated decrease in rate of current activation. Similarly, okadaic acid (100 nM), a protein phosphatase 1/2A inhibitor, selectively occluded the inhibition of the sustained current. 3. The depression of calcium currents by quinpirole (10 nM) was enhanced following intracellular dialysis with 100 microM cyclic adenosine monophosphate (cyclic AMP, 72.8 +/- 9.8% depression), but was not mimicked by the membrane permeant cyclic GMP analogue, Sp-8-bromoguanosine-3',5':cyclic monophosphorothioate (100 microM). 4. Inhibition of calcium currents was only partly attenuated by 100 ms depolarizing prepulses to +100 mV immediately preceding the test pulse. However, following occlusion of the sustained depression with okadaic acid (100 nM) the residual kinetic slowing was reversed in a voltage-dependent manner (P < 0.05). 5. Thus pertussis toxin-sensitive G-proteins liberated upon activation of human D2(short) dopamine receptors inhibited high-threshold calcium currents in two distinct ways. The decrease in rate of calcium current activation involved a voltage-dependent pathway, whereas the sustained inhibition of calcium current involved, in part, the voltage-resistant phosphorylation by cyclic AMP-dependent protein kinases and subsequent dephosphorylation by protein phosphatases 1/2A.
authors.affiliation: Department of Pharmacology, Merck Sharp & Dohme Research Laboratories, Terlings Park, Harlow, Essex.
authors.forename: N A
authors.initials: NA
authors.lastname: Brown
authors.forename: G R
authors.initials: GR
authors.lastname: Seabrook
mesh_terms: D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000633:toxicity; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D000242:Cyclic AMP / Q000494:pharmacology; D004075:Diglycerides / Q000494:pharmacology; D018491:Dopamine Agonists / Q000494:pharmacology; D004558:Electric Stimulation; D004594:Electrophysiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004873:Ergolines / Q000494:pharmacology; D004988:Ethers, Cyclic / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D019319:Okadaic Acid; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000633:toxicity; D037342:Pertussis Toxin; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D054645:Protein Phosphatase 1; D019257:Quinpirole; D017448:Receptors, Dopamine D2 / Q000187:drug effects / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D020866:omega-Conotoxin GVIA
chemical_list: D002121:Calcium Channel Blockers; D015220:Calcium Channels; D004075:Diglycerides; D018491:Dopamine Agonists; D004791:Enzyme Inhibitors; D004873:Ergolines; D004988:Ethers, Cyclic; D010455:Peptides; D017448:Receptors, Dopamine D2; D010566:Virulence Factors, Bordetella; D019319:Okadaic Acid; D019257:Quinpirole; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C038237:1-oleoyl-2-acetylglycerol; D020866:omega-Conotoxin GVIA; D000242:Cyclic AMP; D037342:Pertussis Toxin; D011493:Protein Kinase C; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1; D019204:GTP-Binding Proteins; D002118:Calcium