PHOSPHODIESTERASE INHIBITOR-DEPENDENT INVERSE AGONISM OF AGOUTI-RELATED PROTEIN (AGRP) ON MELANOCORTIN 4 RECEPTOR IN SEA BASS (Dicentrarchus labrax)

The melanocortin 4 receptor (MC4R) is a G-protein coupled receptor mainly expressed in the central nervous system of vertebrates. Activation of the MC4R leads to a decrease in food intake, while inactivating mutations are a genetic cause of obesity. The binding of agoutirelated protein (AGRP) reduces agonist-stimulated cAMP production (competitive antagonist) but also the basal activity of the receptor, as an inverse agonist. Transgenic zebrafish overexpresing AGRP display increased food intake and linear growth, indicative of a physiological role for the melanocortin system in the control of the energy balance in fish. We report on the cloning, pharmacological characterization, tissue distribution and detailed brain mapping of a sea bass (Dicentrarchus labrax) MC4R orthologue. Sea bass MC4R is profusely expressed within food-intake controlling pathways of the fish brain. However, the activity of the melanocortin system during progressive fasting does not depend on the hypothalamic/pituitary proopiomelanocortin (POMC) and MC4R expression, which suggests that sea bass MC4R is constitutively activated and regulated by AGRP binding. We demonstrate that AGRP acts as competitive antagonist and reduces MTII-induced cAMP production. AGRP also decreases the basal activity of the receptor as an inverse agonist. This observation suggests that MC4R is constitutively active and supports the evolutionary conservation of the AGRP/MC4R interactions. The inverse agonism, but not the competitive antagonism, depends on the presence of a phosphodiesterase inhibitor (IBMX). This suggests that inverse agonism and competitive antagonism operate through different intracellular signalling pathways, a view that open up new targets for the treatment of melanocortininduced metabolic syndrome.