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Functional expression of human D 3 dopamine receptors in differentiated neuroblastoma × glioma NG108–15 cells
Author(s) -
Seabrook Guy R.,
Kemp John A.,
Freedman Stephen B.,
Patel Shil,
Sinclair Heather A.,
McAllister George
Publication year - 1994
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/j.1476-5381.1994.tb14746.x
Subject(s) - neuroblastoma , glioma , receptor , dopamine , dopamine receptor , biology , microbiology and biotechnology , neuroscience , cancer research , chemistry , cell culture , genetics
This study describes the depression of calcium currents caused by activation of human D 3 dopamine receptors which have been stably expressed in the neuroblastoma × glioma NG108–15 cell line. Transfected cells, which had been differentiated with prostaglandin E 1 and isobutylmethylxanthine, exclusively expressed D 3 receptor mRNA, which was demonstrated by reverse transcription polymerase chain reaction techniques. Transfected cells had high affinity binding sites for iodosulpiride, with a K d of 0.8 n m and receptor density of 240 fmolmg −1 protein. Calcium currents were recorded using nystatinperforated patch clamp techniques. In contrast to untransfected cells that had been differentiated, high‐threshold calcium currents in differentiated hD 3 ‐NG108–15 cells were depressed by application of dopamine and quinpirole. These responses were abolished by the dopamine receptor antagonist S‐(–)‐sulpiride (1 μ m ), demonstrating that they were caused by the activation of the transfected dopamine receptors. Coupling of human D 3 receptors to calcium currents was sensitive to the action of pertussis toxin, suggesting the involvement of G‐proteins of the G i and/or G o subtype. These results demonstrate that human D 3 receptors represent a functional class of dopamine receptor.