Recovery of β-adrenoceptors and cyclic AMP response after long term treatment of intact heart cells with β-blockers

1 We have studied the recovery of receptor binding and of isoprenaline-stimulated cyclic AMP responses after chronic (2–5 days) exposure of tissue-cultured living rat heart cells to several β-adrenoceptor antagonists. Most experiments were performed with [3H]- (±)-carazolol and [3H]-(±)-CGP 12177, as prototypes of high affinity lipophilic and hydrophilic ligands respectively. 2 Chronic antagonist treatment did not alter the total number of receptors nor did it cause intracellular accumulation of the ligands. 3 At the end of the treatment, radiolabeled antagonists were displaced either by ‘infinite’ dilution of the incubation medium or by competitive displacement with the non-labelled ligand (-)-timolol. In dilution assays dissociation of Carazolol from specific sites was biphasic with t1/2 values of 41 ± 14 and 219 ± 15 min. Dissociation of CGP 12177 was monophasic with t1/2 of 102 ± 2 min. 4 Timolol enhanced the dissociation rates of both radioligands and suppressed the slow phase of Carazolol dissociation. 5 Isoprenaline-stimulated cyclic AMP formation did not recover in parallel with the release of the two antagonists from receptor binding sites. To reach about 80% of control values for receptor availability or cyclic AMP response required 3 h and 24 h washout periods, respectively, after Carazolol (0.2 nm) treatment, or 1.5 and 12 h washout periods after CGP 12177 (4 nm) treatment. Such a ‘decoupling’ effect was not observed during recovery from chronic exposure to the antagonists, timolol and propranolol. 6 We conclude that some antagonists cause a novel form of desensitization that is not linked to their partial agonistic potency. Moreover, Carazolol-type drugs seem to induce an additional isomeric form of the β-receptor that is not recognized by other antagonists. These observations could explain the well known discrepancy between long duration of action and rapid removal from the circulation of several antagonists in current therapeutic use.