Functional coupling of cerebral ?-aminobutyric acid (GABA)(B) receptor with adenylate cyclase system: effect of phaclofen.

Using synaptic membrane from bovine cerebral cortex, effects of ?-aminobutyric acid (GABA), (?)-baclofen, and phaclofen on the cyclic AMP formation mediated by adenylate cyclase were studied. In addition, the binding affinity of phaclofen, a GABA(B) antagonist, to synaptic membrane was compared with those of GABA and (?)-baclofen. GABA and (?)-baclofen, GABA(B) receptor agonists, induced significant inhibitions on the basal and forskolin-stimulated adenylate cyclase activities. Treatment of synaptic membrane with the islet-activating protein, pertussis toxin, completely eliminated the inhibitory effects of GABA and (?)-baclofen on the forskolin-stimulated adenylate cyclase activity. In solubilized fraction of synaptic membrane, the forskolin-stimulated adenylate cyclase was no longer affected by the additions of GABA and (?)-baclofen. Phaclofen displaced 50% of the bound [(3)H](?)-baclofen from synaptic membrane at the concentration of 10(?3) M, and also completely abolished inhibitory effects of GABA and (?)-baclofen on the forskolin-stimulated adenylate cyclase activity. These results suggest that GABA(B) receptor in synaptic membrane of the bovine cerebral cortex may be functionally coupled with adenylate cyclase system via Ni and/or No proteins. The present results also suggest that phaclofen may have selective affinity to the same binding sites as those of GABA(B) receptor agonists such as (?)-baclofen, and induce a suppressive effect on GABA(B) receptor mediated inhibition of adenylate cyclase.