title: Characterization of alpha-adrenoceptors in myometrium of preparturient rats.
abstract: We describe three methods for the quantitative analysis of the alpha-adrenoceptor subtypes in preparturient rat myometrial membrane fractions. A non-subtype-selective antagonist radioligand. [3H]dihydroergocryptine ([3H]DHE), was used to label all of the alpha-receptors. [3H]DHE bound to both alpha 1- and alpha 2-receptors with indistinguishable affinity. Computer modelling of competition curves of unlabeled selective antagonists or agonists was then required in order to determine reliably alpha 1 and alpha 2 affinities and proportions: the alpha 1-receptors represent 45% and the alpha 2-receptors 55% of the entire alpha-receptor population in rat uterus. The second approach involved the administration of phenoxybenzamine (POB) that irreversibly blocks the alpha 1-adrenoceptors. Myometrial membranes obtained from rats 1 h after the administration of varying amounts of POB showed a dose-dependent reduction in specific [3H]DHE binding. This reduction was accompanied by a progressive increase of the value of the dissociation constant. Our data indicate that a dose of 1 mg of POB left the alpha 2-receptors intact while entirely blocking the alpha 1-receptors in rat myometrium. The third approach utilized the selective radioligand antagonists [3H]prazosin ([3H]PRAZ) and [3H]rauwolscine ([3H]RAUW). The results obtained with these radioligands confirmed our observations on the alpha-adrenoceptor subtypes in experiments with [3H]DHE. The results obtained with the 3 methods are in good agreement. Each approach appears valid and applicable to the characterization of alpha 1- and alpha 2-adrenoceptor subtypes in rat uterus, but the method using [3H]PRAZ and [3H]RAUW demonstrates more directly the presence of the two receptor subtypes.
mesh_terms: D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D004088:Dihydroergotoxine / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D007700:Kinetics; D008566:Membranes / Q000378:metabolism; D009215:Myometrium / Q000378:metabolism*; D010643:Phenoxybenzamine / Q000494:pharmacology; D011224:Prazosin / Q000494:pharmacology; D011247:Pregnancy; D051381:Rats; D011919:Rats, Inbred Strains; D011942:Receptors, Adrenergic, alpha / Q000378:metabolism*; D015016:Yohimbine / Q000494:pharmacology
chemical_list: D011942:Receptors, Adrenergic, alpha; D010643:Phenoxybenzamine; D004088:Dihydroergotoxine; D015016:Yohimbine; D011224:Prazosin