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Molecular Pharmacology

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Abstract

Sequestration of the short and long isoforms of dopamine D2 receptors expressed in Chinese hamster ovary cells.

M Itokawa, M Toru, K Ito, H Tsuga, K Kameyama, T Haga, T Arinami and H Hamaguchi
Molecular Pharmacology March 1996, 49 (3) 560-566;
M Itokawa
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M Toru
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K Ito
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H Tsuga
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K Kameyama
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T Haga
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T Arinami
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H Hamaguchi
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Abstract

The short (D2S) and long (D2L) isoforms of dopamine D2 receptors were stably expressed in Chinese hamster ovary cells, and dopamine-induced sequestration was examined by measuring the loss of binding of the hydrophilic ligand [3H]sulpiride from the cell surface. Dopamine treatment of Chinese hamster ovary cells expressing D2S for 30 min at 37 degrees caused a 43.8 +/- 3.4% decrease in [3H]sulpiride binding activity measured by incubation of the treated cells with [3H]sulpiride at 4 degrees for 4 hr after the dopamine was washed out. The half-life of the decrease in binding was estimated to be 18.7 +/- 1.6 min, and the concentration of dopamine giving a half-maximal effect (EC50) was estimated to be 180 +/- 90 nM. The decrease was reversible, and the binding activity was recovered by washing out the dopamine and incubating the cells at 37 degrees for 30 min but was not reversible when the cells were incubated at 4 degrees. The binding activity of [3H]spiperone, a hydrophobic ligand, was not affected by the dopamine treatment under the same experimental conditions. These results indicate that approximately one half of the D2S receptors undergo agonist-induced sequestration, probably endocytosis, in a reversible and temperature-dependent manner. Sequestration of D2L receptors was not as apparent as that of D2S receptors; the decrease in [3H]sulpiride binding activity was 21.6 +/- 0.9% and the rate of the decrease was delayed, with a half-life of 33.2 +/- 7.8 min, although effective concentrations of dopamine were similar, with EC50 = 170 +/- 50 nM. A D2S receptor variant containing a missense mutation changing Ser311 in the third intracellular loop to cysteine was found to be sequestered to a significantly lesser extent than with wild-type D2S receptors. This finding was discussed with respect to the report that this variant gene is found more frequently in schizophrenic patients than in control subjects.

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Molecular Pharmacology
Vol. 49, Issue 3
1 Mar 1996
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Abstract

Sequestration of the short and long isoforms of dopamine D2 receptors expressed in Chinese hamster ovary cells.

M Itokawa, M Toru, K Ito, H Tsuga, K Kameyama, T Haga, T Arinami and H Hamaguchi
Molecular Pharmacology March 1, 1996, 49 (3) 560-566;

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Abstract

Sequestration of the short and long isoforms of dopamine D2 receptors expressed in Chinese hamster ovary cells.

M Itokawa, M Toru, K Ito, H Tsuga, K Kameyama, T Haga, T Arinami and H Hamaguchi
Molecular Pharmacology March 1, 1996, 49 (3) 560-566;
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