The dose- and time-dependent changes of in vivo radioligand binding to the neuronal membrane dopamine transporter (DAT) and vesicular monoamine transporter type 2 (VMAT2) were examined in mouse brain after MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) administrations. Regional brain distribution studies were done in male C57BL/6 mice using simultaneous injections of d - threo -[ 3 H]methylphenidate (DAT) and (+)-α-[ 11 C]dihydrotetrabenazine (VMAT2). Single (55 mg/kg i.p.) or multiple (4 × 10 mg/kg i.p., 1-hour intervals) administration of MPTP caused significant reductions in [ 3 H]methylphenidate and [ 11 C]dihydrotetrabenazine specific striatal binding, measured 14 days later. The single high dose of MPTP produced greater losses of [ 11 C]dihydrotetrabenazine binding than did the multiple MPTP dosing regimen. Using the single high dose of MPTP, changes of in vivo binding of the two radioligands were determined at 1, 3, and 14 days after neurotoxin injection. At 1 day, there are large losses of [ 3 H]methylphenidate binding (DAT) but no changes in [ 11 C]dihydrotetrabenazine binding to the VMAT2 site in the striatum. At 3 and 14 days, there were >50% losses of binding of both bot radioligands, but significantly ( P < 0.001) greater losses of VMAT2 binding of [ 11 C]dihydrotetrabenazine. These studies indicate that the losses of the neuronal membrane and vesicular transporters are not always equal, and do not occur in the same time frame, after administration of the neurotoxin MPTP. Synapse 35:250–255, 2000. © 2000 Wiley-Liss, Inc.