Knockout of adenylyl cyclase isoform 5 or 6 differentially modifies the β₁-adrenoceptor-mediated inotropic response

Although only β₂-adrenergic receptors (βAR) dually couple with stimulatory G protein (G_s) and inhibitory G protein (G_i), inactivation of G_i enhances both β₁AR and β₂AR responsiveness. We hypothesize that G_i restrains spontaneous adenylyl cyclase (AC) activity independent of receptor activation. Subcellular localization of the AC5/6 subtypes varies contributing to the compartmentation of βAR signaling. The primary objectives were to determine: (1) if β₁AR-mediated inotropic responses were dependent upon either AC5 or AC6; (2) if intrinsic G_i inhibition is AC subtype selective and (3) the role of phosphodiesterases (PDE) 3/4 to regulate β₁AR responsiveness. β₁AR-mediated increases in contractile force and cAMP accumulation in cardiomyocytes were measured from wild type, AC5 and AC6 knockout (KO) mice, with or without pertussis toxin (PTX) pretreatment to inactivate G_i and/or after selective inhibition of PDEs 3/4. Noradrenaline potency at β₁ARs was increased in AC6 KO. PDE4 inhibition increased noradrenaline potency in wild type and AC5 KO, but not AC6 KO. PTX increased noradrenaline potency only in wild type but increased the maximal β₁AR response in all mouse strains. PDE3 inhibition increased noradrenaline potency only in AC5 KO that was treated prior with PTX. β₁AR-evoked cAMP accumulation was increased more by PDE4 inhibition than PDE3 inhibition in wild type and AC5 KO that was amplified by G_i inhibition. These data indicate that β₁AR-mediated inotropic responses are not dependent upon either AC5 or AC6 alone. Inactivation of G_i enhanced β₁AR-mediated inotropic responses despite not coupling to G_i, consistent with G_i exerting a tonic receptor independent inhibition upon AC5/6. PDE4 seems the primary regulator of β₁AR signaling through AC6 in wild type. AC6 KO results in a reorganization of β₁AR compartmentation characterized by signaling through AC5 regulated by G_i, PDE3 and PDE4 that maintains normal contractile function.