Identification of adenylate cyclase-coupled beta-adrenergic receptors in the developing mammalian palate.
A direct radioligand binding technique utilizing a beta-adrenergic antagonist [3H]Dihydroalprenolol [( 3H]DHA) was employed in the identification and characterization of fetal palatal beta-adrenergic receptors. [3H]DHA binding was saturable (Bmax 16 fmol/mg protein) with high affinity and an apparent equilibrium dissociation constant (KD) of 1.5 nM. Binding of [3H]DHA was displaced by the competitive beta-adrenergic antagonist propranolol in a concentration-dependent manner. Dissociation kinetic studies demonstrated almost complete reversibility of radioligand binding within 60 min. The functionality of these beta-adrenergic receptors was demonstrated by showing that fetal palatal mesenchymal cells responded to catecholamine agonists with dose-dependent accumulations of intracellular cAMP. This effect could be entirely blocked by the beta-antagonist, propranolol. The relative potency order of catecholamines in eliciting an elevation of cellular cAMP was characteristic of a beta 2-adrenergic receptor-mediated response: (-) isoproterenol greater than (-) epinephrine greater than (-) norepinephrine. In addition, this response was found to be stereospecific with (-) isoproterenol being significantly more potent than (+) isoproterenol. Both the [3H]DHA binding characteristics and the catecholamine sensitivity of fetal palatal tissue support the presence of adenylate cyclase-coupled beta-adrenergic receptors in the developing mammalian secondary palate.
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002395:Catecholamines / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004082:Dihydroalprenolol / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D051379:Mice; D010159:Palate / Q000196:embryology* / Q000378:metabolism; D011433:Propranolol / Q000494:pharmacology; D011943:Receptors, Adrenergic, beta / Q000187:drug effects / Q000378:metabolism*
D002395:Catecholamines; D011943:Receptors, Adrenergic, beta; D004082:Dihydroalprenolol; D011433:Propranolol; D000242:Cyclic AMP; D000262:Adenylyl Cyclases