'Spare' alpha1-adrenergic receptors and the potency of agonists in rat vas deferens

K. P. Minneman, Peter W. Abel

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The existence of 'spare' alpha1-adrenergic receptors in rat vas deferens was examined, directly using radioligand binding assays and contractility measurements. Alpha1-adrenergic receptors in homogenates of rat vas deferens were labeled with [125I]BE 2254 (125IBE). Norepinephrine and other full alpha1-adrenergic receptors agonists were much less potent in inhibiting 125IBE binding than in contracting the vas deferens in vitro. Treatment with 300 nM phenoxybenzamine for 10 min to irreversibly inactive alpha1-adrenergic receptors caused a large decrease in the potency of full angonists in causing contraction of this tissue and a 23-48% decrease in the maximal contraction observed. Using those data, equilibrium constants for activation (K(act) values) of the receptors by agonists were calculated. These K(act) values agreed well with the equilibrium binding constants (K(D) values) determined from displacement of 125IBE binding. The reduction in alpha1-adrenergic receptor density following phenoxybenzamine treatment was determined by Scatchard analysis of specific 125IBE binding sites and compared with the expected reduction (q values) calculated from agonist dose-response curves before and after phenoxybenzamine treatment. Exposure to 300 nM phenoxybenzamine for 10 min resulted in a 39% decreased specific 125IBE binding sites, which did not agree with the 93% decrease expected from the calculated q values. Treatment of vas deferens with a dose of phenoxybenzamine (10 μM for 15 min) that completely abolished the contractile response to alpha1-adrenergic agonists caused an 82% decrease in the density of 125IBE binding sites. Tissues exposed to 300 nM phenoxybenzamine in the presence of 100 μM phentolamine of 3 μM prazosin showed no change in the dose-response curves for agonist-induced contraction or in the density of 125IBE binding sites when compared with controls. This suggests that phenoxybenzamine functionally inactivates alpha1-adrenergic receptors at or near the receptor binding site. These experiments suggest that the potencies of agonists in activating alpha1-adrenergic receptors in rat vas deferens agree well with their potencies in binding to the receptors. The greater potency of agonists in causing contraction may be due to spare receptors in this tissue. The data also demonstrate that phenoxybenzamine irreversibly inactivates alpha1-adrenergic receptors in rat vas deferens, but that the decrease in receptor density is much smaller than that predicted from receptor theory.

Original languageEnglish
Pages (from-to)56-63
Number of pages8
JournalMolecular Pharmacology
Volume25
Issue number1
StatePublished - 1984
Externally publishedYes

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Phenoxybenzamine
Adrenergic Agonists
Vas Deferens
Adrenergic Receptors
Binding Sites
Adrenergic alpha-1 Receptor Agonists
Radioligand Assay
Prazosin
Phentolamine
BE 2254
Norepinephrine

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

'Spare' alpha1-adrenergic receptors and the potency of agonists in rat vas deferens. / Minneman, K. P.; Abel, Peter W.

In: Molecular Pharmacology, Vol. 25, No. 1, 1984, p. 56-63.

Research output: Contribution to journalArticle

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abstract = "The existence of 'spare' alpha1-adrenergic receptors in rat vas deferens was examined, directly using radioligand binding assays and contractility measurements. Alpha1-adrenergic receptors in homogenates of rat vas deferens were labeled with [125I]BE 2254 (125IBE). Norepinephrine and other full alpha1-adrenergic receptors agonists were much less potent in inhibiting 125IBE binding than in contracting the vas deferens in vitro. Treatment with 300 nM phenoxybenzamine for 10 min to irreversibly inactive alpha1-adrenergic receptors caused a large decrease in the potency of full angonists in causing contraction of this tissue and a 23-48{\%} decrease in the maximal contraction observed. Using those data, equilibrium constants for activation (K(act) values) of the receptors by agonists were calculated. These K(act) values agreed well with the equilibrium binding constants (K(D) values) determined from displacement of 125IBE binding. The reduction in alpha1-adrenergic receptor density following phenoxybenzamine treatment was determined by Scatchard analysis of specific 125IBE binding sites and compared with the expected reduction (q values) calculated from agonist dose-response curves before and after phenoxybenzamine treatment. Exposure to 300 nM phenoxybenzamine for 10 min resulted in a 39{\%} decreased specific 125IBE binding sites, which did not agree with the 93{\%} decrease expected from the calculated q values. Treatment of vas deferens with a dose of phenoxybenzamine (10 μM for 15 min) that completely abolished the contractile response to alpha1-adrenergic agonists caused an 82{\%} decrease in the density of 125IBE binding sites. Tissues exposed to 300 nM phenoxybenzamine in the presence of 100 μM phentolamine of 3 μM prazosin showed no change in the dose-response curves for agonist-induced contraction or in the density of 125IBE binding sites when compared with controls. This suggests that phenoxybenzamine functionally inactivates alpha1-adrenergic receptors at or near the receptor binding site. These experiments suggest that the potencies of agonists in activating alpha1-adrenergic receptors in rat vas deferens agree well with their potencies in binding to the receptors. The greater potency of agonists in causing contraction may be due to spare receptors in this tissue. The data also demonstrate that phenoxybenzamine irreversibly inactivates alpha1-adrenergic receptors in rat vas deferens, but that the decrease in receptor density is much smaller than that predicted from receptor theory.",
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