Expression and modulation of 5-hydroxytryptamine(1A) receptors in P11 cells

J. G. Hensler, L. S. Cervera, H. A. Miller, J. Corbitt

Research output: Contribution to journalArticle

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Abstract

P11 cells were transfected with DNA for the human 5- hydroxytryptamine(1A) (5-HT(1A)) receptor. These cells stably expressed the 5-HT(1A) receptor coupled to the inhibition of adenylyl cyclase, and not to the stimulation of phosphoinositide hydrolysis. Homologous and heterologous regulation of the 5-HT(1A) receptor was studied in this cell system. Pretreatment of P11-5HT(1A) cells with the 5-HT1 receptor agonist 5- carboxamidotryptamine (5-CT) resulted in a 3-fold increase in both basal and forskolin-stimulated cAMP accumulation, and desensitization of the 5-HT(1A) receptor as indicated by a decrease in the potency of 8- hydroxydipropylaminotetralin (8-OH-DPAT) to inhibit forskolin-stimulated cAMP accumulation (vehicle-treated cells: EC50 = 2.3 ± 0.6 nM; 5-CT-treated cells: 9.9 ± 0.4 nM). The sensitization of adenylyl cyclase as a result of chronic agonist exposure was prevented by the 5-HT(1A) antagonist WAY100635, which indicated that the effect of 5-CT pretreatment on basal and forskolin- stimulated cAMP accumulation was mediated by 5-HT(1A) receptor activation. Pretreatment of cells with pertussis toxin abolished the inhibition of forskolin-stimulated cAMP accumulation mediated by 5-HT(1A) receptor activation and prevented the sensitization of adenylyl cyclase as a result of chronic 5-HT(1A) receptor agonist exposure. Pretreatment of P11-5HT(1A) cells with the phorbol ester, phorbol 12-myristate 13-acetate (PMA), also resulted in desensitization of the 5-HT(1A) receptor, as indicated by a marked decrease in the potency and intrinsic activity of 8-OH-DPAT. No change in the binding characteristics (i.e., K(d) or B(max)) of [3H]8-OH-DPAT to 5-HT(1A) receptor sites was observed after 5-CT or PMA treatments. Activation of alpha-1 adrenergic receptors, but not 5-HT(2A) receptors, had effects on 5- HT(1A) receptor responsiveness similar to those seen with PMA pretreatment. In P11-5HT(1A) cells, homologous regulation of the 5-HT(1A) receptor was characterized by sensitization of adenylyl cyclase and a decrease in agonist potency, whereas heterologous regulation of the 5-HT(1A) receptor was characterized by a greater decrease in agonist potency, as well as a marked decrease in intrinsic activity.

Original languageEnglish (US)
Pages (from-to)1138-1145
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume278
Issue number3
StatePublished - 1996

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Receptor, Serotonin, 5-HT1A
Colforsin
Adenylyl Cyclases
8-Hydroxy-2-(di-n-propylamino)tetralin
Acetates
Adrenergic alpha-1 Receptors
Serotonin 5-HT1 Receptors
Serotonin 5-HT1 Receptor Agonists
Receptor, Serotonin, 5-HT2A
Serotonin Antagonists
Pertussis Toxin
Phorbol Esters
Phosphatidylinositols
Hydrolysis

ASJC Scopus subject areas

  • Pharmacology

Cite this

Hensler, J. G., Cervera, L. S., Miller, H. A., & Corbitt, J. (1996). Expression and modulation of 5-hydroxytryptamine(1A) receptors in P11 cells. Journal of Pharmacology and Experimental Therapeutics, 278(3), 1138-1145.

Expression and modulation of 5-hydroxytryptamine(1A) receptors in P11 cells. / Hensler, J. G.; Cervera, L. S.; Miller, H. A.; Corbitt, J.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 278, No. 3, 1996, p. 1138-1145.

Research output: Contribution to journalArticle

Hensler, JG, Cervera, LS, Miller, HA & Corbitt, J 1996, 'Expression and modulation of 5-hydroxytryptamine(1A) receptors in P11 cells', Journal of Pharmacology and Experimental Therapeutics, vol. 278, no. 3, pp. 1138-1145.
Hensler, J. G. ; Cervera, L. S. ; Miller, H. A. ; Corbitt, J. / Expression and modulation of 5-hydroxytryptamine(1A) receptors in P11 cells. In: Journal of Pharmacology and Experimental Therapeutics. 1996 ; Vol. 278, No. 3. pp. 1138-1145.
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N2 - P11 cells were transfected with DNA for the human 5- hydroxytryptamine(1A) (5-HT(1A)) receptor. These cells stably expressed the 5-HT(1A) receptor coupled to the inhibition of adenylyl cyclase, and not to the stimulation of phosphoinositide hydrolysis. Homologous and heterologous regulation of the 5-HT(1A) receptor was studied in this cell system. Pretreatment of P11-5HT(1A) cells with the 5-HT1 receptor agonist 5- carboxamidotryptamine (5-CT) resulted in a 3-fold increase in both basal and forskolin-stimulated cAMP accumulation, and desensitization of the 5-HT(1A) receptor as indicated by a decrease in the potency of 8- hydroxydipropylaminotetralin (8-OH-DPAT) to inhibit forskolin-stimulated cAMP accumulation (vehicle-treated cells: EC50 = 2.3 ± 0.6 nM; 5-CT-treated cells: 9.9 ± 0.4 nM). The sensitization of adenylyl cyclase as a result of chronic agonist exposure was prevented by the 5-HT(1A) antagonist WAY100635, which indicated that the effect of 5-CT pretreatment on basal and forskolin- stimulated cAMP accumulation was mediated by 5-HT(1A) receptor activation. Pretreatment of cells with pertussis toxin abolished the inhibition of forskolin-stimulated cAMP accumulation mediated by 5-HT(1A) receptor activation and prevented the sensitization of adenylyl cyclase as a result of chronic 5-HT(1A) receptor agonist exposure. Pretreatment of P11-5HT(1A) cells with the phorbol ester, phorbol 12-myristate 13-acetate (PMA), also resulted in desensitization of the 5-HT(1A) receptor, as indicated by a marked decrease in the potency and intrinsic activity of 8-OH-DPAT. No change in the binding characteristics (i.e., K(d) or B(max)) of [3H]8-OH-DPAT to 5-HT(1A) receptor sites was observed after 5-CT or PMA treatments. Activation of alpha-1 adrenergic receptors, but not 5-HT(2A) receptors, had effects on 5- HT(1A) receptor responsiveness similar to those seen with PMA pretreatment. In P11-5HT(1A) cells, homologous regulation of the 5-HT(1A) receptor was characterized by sensitization of adenylyl cyclase and a decrease in agonist potency, whereas heterologous regulation of the 5-HT(1A) receptor was characterized by a greater decrease in agonist potency, as well as a marked decrease in intrinsic activity.

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