Role of muscarinic M2 receptors in regulating beta-adrenergic responsiveness in maturing rabbit airway smooth muscle.

Abstract

Muscarinic M2 and M3 receptor subtypes have been pharmacologically distinguished in airway smooth muscle. Whereas M3 receptors have been associated with smooth muscle contraction, M2 receptors have been implicated in Gi protein-coupled inhibition of adenylyl cyclase. To determine whether the role of M2 receptors varies with age in tracheal smooth muscle (TSM), dose-dependent relaxation responses to isoproterenol were compared in TSM isolated from 3-day-old and adult rabbits precontracted with acetylcholine (ACh) in the absence (control) and presence of an M2 receptor antagonist (gallamine or methoctramine). From sustained half-maximal ACh contractions, adult TSM were 5.6-fold less sensitive than 3-day-old tissues to isoproterenol-induced relaxation. Furthermore, the magnitude of muscarinic functional antagonism of isoproterenol-mediated TSM relaxation, assessed by varying the initial degree of ACh-induced contraction, significantly increased with age. In gallamine- and methoctramine-treated tissues, the relaxation-response curves to isoproterenol were shifted to the left in both 3-day-old and adult TSM. In contrast, pretreatment with either M2 receptor antagonist had no significant effect on the magnitude of muscarinic functional antagonism at either age. Moreover, Western blot analysis of G alpha i common and specific subunit expression in TSM membranes demonstrated qualitatively similar levels in 3-day-old and adult TSM. Collectively, these findings provide new evidence that 1) there exist inherent age-dependent differences in both the airway relaxant responsiveness to beta-adrenoceptor stimulation and muscarinic functional antagonism of beta-adrenergic relaxation, and 2) the latter are attributed to mechanisms other than ontogenetic alteration in M2 receptor function or Gi protein expression in maturing rabbit TSM.