Dopamine (DA) D2 receptor supersensitivity has been linked to an increase in the density of the D2 high-affinity state as measured in vitro. The two- affinity-state model of the D2 receptor predicts that the ex vivo specific binding of [11C]-(+)-PHNO, an agonist radiotracer thought to bind selectively to the high-affinity state in vivo, should be increased in animal models that display in vitro increases in the proportion of receptors in the D2 high-affinity state. Here, we test this hypotheses by comparing the ex vivo SBR of [11C]-(+)-PHNO with that of the antagonist radiotracer [3H]-raclopride in three dopaminergically supersensitive rat models-AMPH-sensitized rats, rats withdrawn from chronic ethanol, and unilaterally 6-OHDA-lesioned rats-using ex vivo dual-radiotracer biodistribution studies. We find that in AMPH-sensitized rats and rats withdrawn from chronic ethanol treatment, models that exhibited approximately 4-fold increases in the D2 high-affinity state in vitro, the SBRs of [11C]-(+)-PHNO and [3H]-raclopride are unchanged relative to control rats. In unilaterally 6-OHDA-lesioned rats, we find that the increase in [11C]-(+)-PHNO SBR is no different than that observed for the antagonist radiotracer [3H]-raclopride (54% +/- 16% and 52% +/- 14%, respectively). In addition, the effect of acute AMPH pretreatment (4 mg/kg, i.v.) on the SBRs of [11C]-(+)-PHNO and [3H]-raclopride is equivalent in AMPH-sensitized (-38% +/- 12% and -36% +/- 8%, respectively) and in control rats (-40% +/- 11% and -38% +/- 7%). These data emphasize a significant discrepancy between in vitro and in vivo measures of D2 agonist binding, indicating that the two-affinity-state model of the D2 receptor may not apply veridically to living systems. The potential implications of this discrepancy are discussed.