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Molecular Pharmacology

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Abstract

Serotonin-2 receptors coupled to phosphoinositide hydrolysis in a clonal cell line.

K J Ivins and P B Molinoff
Molecular Pharmacology May 1990, 37 (5) 622-630;
K J Ivins
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Abstract

A permanent line of cells has been established from the transplantable rat pituitary tumor 7315a. P11 cells have been cloned repeatedly, and after more than 60 passages their growth and characteristics are stable. Results of radioligand binding studies with 125I-lysergic acid diethylamide (125I-LSD) indicate that P11 cells express serotonin-2 (5-HT2) receptors. Analysis of the binding of 125I-LSD to membranes prepared from P11 cells revealed the presence of a single class of high affinity sites (Kd = 1.6 nM; Bmax = 211 fmol/mg of protein). The pharmacological profile of the inhibition of the binding of 125I-LSD by a panel of drugs was consistent with the expected profile of these drugs at 5-HT2 receptors. The affinity of the site for serotonin was in the low micromolar range and was decreased by GTP. Phosphoinositide hydrolysis in P11 cells, measured in the presence of lithium, was stimulated by serotonin. Increasing concentrations of the 5-HT2-selective antagonist ketanserin blocked phosphoinositide hydrolysis stimulated by serotonin, and Schild analysis was consistent with a simple competitive interaction. The Ki for ketanserin derived from Schild analysis was comparable to the Ki for ketanserin at the binding site for 125I-LSD. These results suggest that stimulation of phosphoinositide hydrolysis in P11 cells by serotonin is mediated by 5-HT2 receptors. Pretreatment of P11 cells with pertussis toxin caused ADP-ribosylation of Gi and Go, but did not affect the ability of serotonin to stimulate phosphoinositide hydrolysis. Therefore, the guaninine nucleotide-binding protein involved in the coupling of 5-HT2 receptors to phospholipase C in P11 cells is unlikely to be either Gi or Go. P11 cells expressing 5-HT2 receptors coupled to phosphoinositide hydrolysis will be a useful model system for future studies of the regulation and function of 5-HT2 receptors on cultured cells.

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Molecular Pharmacology
Vol. 37, Issue 5
1 May 1990
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Abstract

Serotonin-2 receptors coupled to phosphoinositide hydrolysis in a clonal cell line.

K J Ivins and P B Molinoff
Molecular Pharmacology May 1, 1990, 37 (5) 622-630;

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Abstract

Serotonin-2 receptors coupled to phosphoinositide hydrolysis in a clonal cell line.

K J Ivins and P B Molinoff
Molecular Pharmacology May 1, 1990, 37 (5) 622-630;
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