Biochemical characteristics of imidazoline specific binding sites from the human brainstem were further investigated using [3H]idazoxan as radiolabeled ligand. The study of the interaction of [3H]idazoxan binding sites with heparin and lectins (soybean and lentil lectin) confirm the heterogeneity of these sites in the human brain. In fact, about 10-15% of [3H]idazoxan binding sites were retained by each of the three supports used, leading to the hypothesis that two populations of sites, with different biochemical characteristics, coexist in this tissue. A small proportion of [3H]idazoxan binding sites was retained on an affinity chromatography support consisting of a clonidine-derived Pharmalink column. The binding activity of these clonidine-eluted sites was markedly and dose-dependently improved by the addition of 'treated fall-through' fraction from the same column. On the other hand, this 'treated fall-through' fraction inhibited the binding activity detected in the solubilized human brainstem membranes. These results also suggest the existence of heterogeneous imidazoline specific binding sites in the human brainstem and the existence of endogenous factors able to discriminate between them.