Relationship of Mu opioid receptor binding to activation of G-proteins in specific rat brain regions

This study investigated the relationship between mu receptor binding and mu agonist activation of G-proteins in the rat brain. To directly compare ago nist potencies in receptor binding (K-i values) and G-protein activation (K -s values), both agonist-stimulated [S-35]guanosine-5'-O-(gamma-thio)-triph osphat ([S-35]GTP gamma S) and [H-3]naloxone binding assays were conducted under identical conditions, using the full mu agonist [d-Ala(2), N-Me-4, Gl y(5)-ol]-enkephalin (DAMGO). DAMGO exhibited biphasic competition of [H-3]n aloxone binding and stimulation of [S-35]GTP gamma S binding in most region s. Whereas the high affinity component represented a large percentage (50-8 0%) of total receptor sites, the high-affinity component of DAMGO-stimulate d [S-35]GTP gamma S binding was much lower, <30% of the total, and in most regions significant stimulation of [S-35]GTP gamma S binding did not occur until the high-affinity binding sites were completely occupied. Moreover, t he low-affinity potencies for DAMGO in receptor binding and G-protein activ ation were the same across different regions. Receptor-transducer amplifica tion factors were calculated by the ratio of the apparent B-max of net agon ist-stimulated [S-35]GTP gamma S binding to the B-max of receptor binding. Amplification factors for the nine regions examined were relatively high an d varied significantly across regions, from a ratio of 8 in the thalamus to 38 in the cortex, suggesting that the efficiency of mu opioid receptor cou pling to G-proteins varies across brain regions. BIOCHEM PHARMACOL 59;11:13 95-1401, 2000. (C) 2000 Elsevier Science Inc.