ACTIVATION OF MITOGEN-ACTIVATED PROTEIN-KINASES BY STIMULATION OF THECENTRAL CANNABINOID RECEPTOR CB1

The G-protein-coupled central cannabinoid receptor (CB1) has been show n to be functionally associated with several biological responses incl uding inhibition of adenylate cyclase, modulation of ion channels and induction of the immediate-early gene Krox-24. Using stably transfecte d Chinese Hamster Ovary cells expressing human CB1 we show here that c annabinoid treatment induces both phosphorylation and activation of mi togen-activated protein (MAP) kinases, and that these effects are inhi bited by SR 141716A, a selective CB1 antagonist. The two p42 and p44 k Da MAP kinases are activated in a time- and dose-dependent manner. The rank order of potency for the activation of MAP kinases with various cannabinoid agonists is CP-55940 > Delta(9)-tetrahydrocannabinol > WIN 55212.2, in agreement with the pharmacological profile of CB1. The ac tivation of MAP kinases is blocked by pertussis toxin but not by treat ment with hydrolysis-resistant cyclic AMP analogues. This suggests tha t the signal transduction pathway between CB1 and MAP kinases involves a pertussis-toxin-sensitive GTP-binding protein and is independent of cyclic AMP metabolism. This coupling of CB1 subtype and mitogenic sig nal pathway, also observed in the human astrocytoma cell line U373 MG, may explain the mechanism of action underlying cannabinoid-induced Kr ox-24 induction.