Abstract Purpose To investigate the role of cation transporters (OCTs, MATEs) in the renal and hepatic disposition of the radiolabeled antiemetic drug [¹¹C]metoclopramide in mice with PET. Methods PET was performed in wild-type mice after administration of an intravenous microdose (<1 μg) of [¹¹C]metoclopramide without and with co-administration of either unlabeled metoclopramide (5 or 10 mg/kg) or the prototypical cation transporter inhibitors cimetidine (150 mg/kg) or sulpiride (25 mg/kg). [¹¹C]Metoclopramide PET was also performed in wild-type and Slc22a1/2^(−/−) mice. Radiolabeled metabolites were measured at 15 min after radiotracer injection and PET data were corrected for radiolabeled metabolites. Results [¹¹C]Metoclopramide was highly metabolized and [¹¹C]metoclopramide-derived radioactivity was excreted into the urine. The different investigated treatments decreased (~2.5-fold) the uptake of [¹¹C]metoclopramide from plasma into the kidney and liver, inhibited metabolism and decreased (up to 3.8-fold) urinary excretion, which resulted in increased plasma concentrations of [¹¹C]metoclopramide. Kidney and liver uptake were moderately (~1.3-fold) reduced in Slc22a1/2^(−/−) mice. Conclusions Our results suggest a contribution of OCT1/2 to the kidney and liver uptake and of MATEs to the urinary excretion of [¹¹C]metoclopramide in mice. Cation transporters may contribute, next to variability in the activity of metabolizing enzymes, to variability in metoclopramide pharmacokinetics and side effects.