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PMID:7512953

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Citation

Chio, CL, Drong, RF, Riley, DT, Gill, GS, Slightom, JL and Huff, RM (1994) D4 dopamine receptor-mediated signaling events determined in transfected Chinese hamster ovary cells. J. Biol. Chem. 269:11813-9

Abstract

A Chinese hamster ovary (CHO) cell line stably expressing a recombinant human D4 dopamine receptor made from a synthetic gene has been used to determine potential D4-mediated signaling events. We designed and synthesized a modified gene coding for a human D4 receptor with reduced G + C content but unaltered encoded amino acids. Stable expression of this gene was obtained in two cell lines, inducible expression in CHO lacI cells and constitutive expression in HEK293 cells. In CHO lacI cells induced to express D4 receptors but not in uninduced cells, dopamine and quinpirole inhibit forskolin-stimulated cAMP accumulation and potentiate ATP-stimulated [3H]arachidonic acid release through a mechanism that requires protein kinase C but is unaffected by membrane-soluble cAMP analogs. In addition, D4 receptor activation causes an increase in the rate of extracellular acidification measured by microphysiometry. This response is unaffected by protein kinase C down-regulation but is inhibited by removal of extracellular sodium and inhibitors of NaH-1 exchange, suggesting the involvement of a Na+/H+ exchanger. All responses are blocked by clozapine and are sensitive to pertussis toxin. D4 receptors, like other G(i)/G(o)-linked receptors, mediate multiple signaling events, and the pathways activated are similar to those used by D2 and D3 receptors expressed in similar cells.

Links

PubMed

Keywords

Amiloride/analogs & derivatives; Amiloride/pharmacology; Animals; Arachidonic Acid/metabolism; Base Sequence; Butaclamol/pharmacology; CHO Cells; Cloning, Molecular; Clozapine/pharmacology; Cricetinae; Cyclic AMP/analogs & derivatives; Cyclic AMP/metabolism; Cyclic AMP/pharmacology; DNA/chemical synthesis; Dopamine/pharmacology; Dopamine Antagonists; Ergolines/pharmacology; Genes, Synthetic; Haloperidol/pharmacology; Humans; Kinetics; Molecular Sequence Data; Pertussis Toxin; Protein Kinase C/antagonists & inhibitors; Protein Kinase C/metabolism; Quinpirole; Raclopride; Receptors, Dopamine/biosynthesis; Receptors, Dopamine/physiology; Receptors, Dopamine D2; Receptors, Dopamine D4; Recombinant Proteins/antagonists & inhibitors; Recombinant Proteins/biosynthesis; Recombinant Proteins/metabolism; Salicylamides/pharmacology; Signal Transduction/drug effects; Sodium-Hydrogen Antiporter/antagonists & inhibitors; Sodium-Hydrogen Antiporter/metabolism; Transfection; Virulence Factors, Bordetella/pharmacology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:DRD4

located_in

GO:0016020: membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:DRD4

involved_in

GO:0007195: adenylate cyclase-inhibiting dopamine receptor signaling pathway

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DRD4

part_of

GO:0016020: membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:DRD4

involved_in

GO:0032417: positive regulation of sodium:proton antiporter activity

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DRD4

involved_in

GO:0050482: arachidonic acid secretion

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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