In mammals, P-glycoprotein immunostaining at the blood-brain barrier has implicated the multidrug pump in the restricted movement of many cytotoxic agents into the central nervous system (CNS). Since many insects require a sophisticated blood-brain barrier system to protect their CNS from plant-derived neurotoxins, we have investigated the possibility that a P-glycoprotein homolog constitutes a component of the insect blood-brain barrier. We have used the nicotine-resistant tobacco hornworm (Manduca sexta) to address this issue. Manduca has been previously shown, in physiological studies, to have an alkaloid (nicotine/morphine/atropine) pump at its excretory malpighian tubules. We show (1) that the tubules are P-glycoprotein immunopositive, (2) that Manduca has a metabolic blood-brain barrier for nicotine, (3) that the barrier co-localizes with P-glycoprotein immunostaining, and (4) that detoxifying enzymes as well as the nicotine pump are likely to account for the metabolic blood-brain barrier to nicotine. These findings may provide insights on two major fronts, the troublesome problem of multi-insecticide resistance, a phenomenon that parallels multidrug resistance in tumor cells, and the problem of tolerance to addictive neuroactive drugs like nicotine or morphine.