Comparison of the interaction of agmatine and crude methanolic extracts of bovine lung and brain with α2‐adrenoceptor binding sites

1. In the present study we have evaluated whether alpha 2-adrenoceptor binding sites on bovine cerebral cortex membranes labelled by [3H]-clonidine, [3H]-idazoxan and [3H]-RX-821002 can distinguish between known agonists and antagonists. This model has then been used to compare the binding profiles of the putative non-catecholamine, clonidine-displacing substance (CDS), agmatine and crude methanolic extracts of bovine lung and brain. 2. Saturation studies carried out in the presence and absence of noradrenaline, 10 mumol 1(-1), revealed that the maximum number of binding sites on bovine cerebral cortex membranes for [3H]-idazoxan and [3H]-RX-821002 were approximately 60-80% greater than those for [3H]-clonidine (62.6 fmol mg-1 protein). Rauwolscine, the selective alpha 2-adrenoceptor antagonist, was approximately 100 fold more potent against each of the ligands than the selective alpha 1-adrenoceptor diastereoisomer, corynanthine. Also, the pKi value for the selective alpha 1-adrenoceptor prazosin against each ligand was less than 6. 3. Adrenaline, UK-14034, rauwolscine, corynanthine, RX-811059 and prazosin produced concentration-dependent inhibition of binding of all three 3H-ligands. The agonists, adrenaline and UK-14304, were approximately 5 and 10 fold less potent against [3H]-idazoxan and [3H]-RX-821002, respectively, than against [3H]-clonidine. In marked contrast, the antagonists, rauwolscine, corynanthine, RX-811059 and prazosin exhibited a different profile, being approximately 2-3 fold more potent against sites labelled by [3H]-RX-821002 and [3H]-idazoxan compared to sites labelled by [3H]-clonidine. 4. Agmatine and histamine produced a concentration-dependent displacement of [3H]-clonidine, [3H]-idazoxan and [3H]-RX-821002 binding to bovine cerebral cortex membranes. The pKi values for agmatine and histamine were independent of the 3H-ligand employed, approximately 4.8 and 4.5,respectively.5. Crude methanolic extracts of bovine brain and lung produced a concentration-dependent inhibition of [3H]-clonidine binding to bovine cerebral cortex membranes (>90%). Based on the volume of the extract that caused 50% inhibition of [3H]-clonidine binding, bovine lung contains 3 fold more CDS than bovine brain. Both extracts were at least 5 fold more potent against a2-adrenoceptor sites labelled by[3H]-clonidine than those labelled by [3H]-idazoxan and [3H]-RX-821002.6. All three 3H-ligands label the same population of alpha2-adrenoceptor binding sites on bovine cerebral cortex membranes, but [3H]-clonidine appears to label selectively the 'agonist' state of the sites: for which known agonists, adrenaline and UK-14304, exhibit a higher affinity. Our results indicate that neither agmatine nor histamine can account for the CDS activity present in crude extracts of bovine brain and lung. Moreover, these extracts appear to possess a binding profile similar to that of adrenaline and UK-14304, suggesting that they may possess agonist activity.