There are several inconsistencies in the literature as regards the characteristics of benzamide and butyrophenone binding to dopamine D2-like receptors. The variations observed in Bmax, Kd and Ki values have led to hypotheses, such as the existence of a specific "benzamide binding site' and that dopamine D2 receptors exist in a monomer-dimer equilibrium, where benzamides are supposed to bind receptor monomers and butyrophenones receptor dimers. We have previously suggested that the discrepant results may instead be related to methodological difficulties associated with the use of very high-affinity radioligands (e.g. ligand depletion and failure to achieve equilibrium). The present study was designed to reinvestigate and critically reevaluate the binding characteristics of [3H]spiperone, [3H]nemonapride, [125I](S)-3-iodo-N-[(1-ethyl-2-pyrrolidinyl)methyl]-5,6- dimethylsalicylamide ([125I]NCQ-298) and [3H]raclopride to cloned human dopamine D2A and rat striatal dopamine D2 receptors in order to establish whether they label the same receptor population. We found that the Kd values of [3H]spiperone, [125I]NCQ-298 and [3H]nemonapride were about 20 pM and that of [3H]raclopride about 1 nM. We did not find any significant differences between the Bmax values determined with the various radioligands. Furthermore, the Ki values of spiperone and NCQ-298 (derived from cross-competition studies) for dopamine D2 receptors labelled with either [3H]spiperone or [125I]NCQ-298 were in good agreement with the corresponding Kd values. In conclusion, our results clearly demonstrate that when studied under correct experimental conditions, all four radioligands label an identical receptor population.