Regulation of the beta-Adrenergic Receptor …
Hoe, L. E. S., et al. (2019). "Regulation of the beta-Adrenergic Receptor Signaling Pathway in Sustained Ligand-Activated Preconditioning." Journal of Pharmacology and Experimental Therapeutics 369(1): 37-46.
Sustained ligand-activated preconditioning (SLP), induced with chronic opioid receptor (OR) agonism, enhances tolerance to ischemia/reperfusion injury in young and aged hearts. Underlying mechanisms remain ill-defined, although early data implicate phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) during the induction phase, and beta(2)-adrenoceptor (beta(2)-AR), G(s) alpha subunit (G alpha(s)), and protein kinase A (PKA) involvement in subsequent cardioprotection. Here, we tested for induction of a protective beta(2)-AR/G alpha(s)/PKA signaling axis with SLP to ascertain whether signaling changes were PI3K-dependent (by sustained cotreatment with wortmannin), and whether the downstream PKA target Rho kinase (ROCK) participates in subsequent cardioprotection (by acute treatment with fasudil). A protected phenotype was evident after 5 days of OR agonism (using morphine) in association with increased membrane versus reduced cytosolic levels of total and phosphorylated beta(2)-ARs; increased membrane and cytosolic expression of 52 and 46 kDa G alpha(s) isoforms, respectively; and increased phosphorylation of PKA and Akt. Nonetheless, functional sensitivities of beta(2)-ARs and adenylyl cyclase were unchanged based on concentration-response analyses for formoterol, fenoterol, and 6-[3-(dimethylamino)propionyl]-forskolin. Protection with SLP was not modified by ROCK inhibition, and changes in beta(2)-AR, G alpha(s), and PKA expression appeared insensitive to PI3K inhibition, although 5 days of wortmannin alone exerted unexpected effects on signaling (also increasing membrane beta(2)-AR and PKA expression/phosphorylation and G alpha(s) levels). In summary, sustained OR agonism upregulates cardiac membrane beta(2)-AR expression and phosphorylation in association with increased G alpha(s) subtype levels and PKA phosphorylation. While Akt phosphorylation was evident, PI3K activity appears nonessential to OR upregulation of the beta(2)-AR signal axis. This opioidergic remodeling of beta(2)-AR signaling may explain beta(2)-AR, G alpha(s), and PKA dependence of SLP protection.