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Molecular Pharmacology

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Abstract

Hamster alpha 1B-adrenergic receptor directly activates Gs in the transfected Chinese hamster ovary cells.

K Horie, H Itoh and G Tsujimoto
Molecular Pharmacology September 1995, 48 (3) 392-400;
K Horie
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H Itoh
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G Tsujimoto
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Abstract

A prototypic Ca(2+)-mobilizing hormone receptor, alpha 1-adrenergic receptor (alpha 1AR), stimulates cAMP accumulation. The mechanism underlying this phenomenon was previously suggested to be secondary to phosphatidylinositol hydrolysis-protein kinase C activation in some cells. We transfected Chinese hamster ovary (CHO)-K1 cells with hamster alpha(1B)AR cDNA and isolated cells stably expressing alpha(1B)AR (CHO alpha 1B cells). We investigated the molecular mechanism underlying the alpha 1AR-mediated cAMP production in the CHO alpha 1B cells. Norepinephrine (NE) stimulated intracellular calcium mobilization and cAMP production through alpha(1B)AR. Pretreatment with a phospholipase C inhibitor, U-73,122 (10 microM), abolished the NE-induced intracellular calcium response, whereas it did not affect the NE-stimulated cAMP production. Treatment with various agents (protein kinase C inhibitors, calcium ionophore, cyclo-oxygenase inhibitor, or pertussis toxin) had little effect on the NE-induced cAMP production. The parent CHO and CHO alpha 1B cells contained similar amounts of Gs alpha (42 and 45 kDa, respectively), as detected with immunoblot analysis, and exhibited similar extents of cAMP synthesis with cholera toxin and forskolin. Adenylyl cyclase activity in the CHO alpha 1B cell membranes was also enhanced by NE. Furthermore, incubation of CHO alpha 1B cell membranes with antiserum directed against the carboxyl-terminal portion of Gs alpha inhibited the NE-stimulated adenylyl cyclase activity. Taken together, the results indicate that the alpha(1B)AR-mediated cAMP synthesis in CHO alpha 1B cells reflects direct stimulation of Gs-adenylyl cyclase. Therefore, the alpha 1AR-stimulated cAMP production observed in some native tissues may involve the multiple mechanisms of the direct activation of Gs-adenylyl cyclase and a secondary effect through activation of phosphatidylinositol hydrolysis.

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Molecular Pharmacology
Vol. 48, Issue 3
1 Sep 1995
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Abstract

Hamster alpha 1B-adrenergic receptor directly activates Gs in the transfected Chinese hamster ovary cells.

K Horie, H Itoh and G Tsujimoto
Molecular Pharmacology September 1, 1995, 48 (3) 392-400;

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Abstract

Hamster alpha 1B-adrenergic receptor directly activates Gs in the transfected Chinese hamster ovary cells.

K Horie, H Itoh and G Tsujimoto
Molecular Pharmacology September 1, 1995, 48 (3) 392-400;
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