Physiological evidence for β3-adrenoceptor in frog (Rana esculenta) heart

β3-Adrenergic receptors (ARs) have been recently identified in mammalian hearts where, unlike β1- and β2-ARs, induce cardio-suppressive effects. The aim of this study was to describe β3-AR role in the frog (Rana esculenta) heart and to examine its signal transduction pathway. The presence of β3-AR, by using Western blotting analysis, has been also identified. BRL37344, a selective β3-AR agonist, induced a dose-dependent negative inotropic effect at concentrations from 10−12 to 10−6 M. This effect was not modified by nadolol (β1/β2-AR antagonist) and by phentolamine (α-AR antagonist), but it was suppressed by the β3-AR-specific antagonist SR59230 and by exposure to the Gi/o proteins inhibitor Pertussis Toxin. In addition, the involvement of EE-NOS-cGMP-PKG/PDE2 pathway in the negative inotropism of BRL37344 has been assessed. BRL37344 treatment induced eNOS and Akt phosphorylation as well as an increase of cGMP levels. β3-ARs activation induce a non-competitive antagonism against ISO stimulation which disappeared in presence of PKG and PDE2 inhibition. Taken together our findings provide, for the first time in the frog, a role for β3-ARs in the cardiac performance modulation which involves Gi/o protein and occurs via an EE-NO-cGMP-PKG/PDE2 cascade.

Research highlights
► β3-Adrenoceptor (β3-AR) induces a cardio-suppressive action in frog.
► β3-AR expression in the frog ventricle has been determined.
► The decrease of cardiac contractility occurs via the Gi-eNOS-cGMP-PDE2/PKG pathway.
► β3-AR activation induces a non-competitive antagonism against ISO stimulation.