AbstractPollination is important for both agriculture and biodiversity. For a significant number of plants, this process is highly and sometimes exclusively, dependent on the pollination activity of honeybees. The large numbers of honeybee colony losses reported in recent years have been attributed to colony collapse disorder. Various hypotheses, including pesticide overuse, have been suggested to explain the disorder. Using theXenopusoocytes expression system and two microelectrode voltage-clamp, we report the functional expression and the molecular, biophysical and pharmacological characterization of the western honeybee’s sodium channel (Apis MelliferaNaV1). The NaV1 channel is the primary target for pyrethroid insecticides in insect pests. We further report that the honeybee’s channel is also sensitive to permethrin and fenvalerate, respectively type I and type II pyrethroid insecticides. Molecular docking of these insecticides revealed a binding site that is similar to sites previously identified in other insects. We describein vitroandin silicotools that can be used to test chemical compounds. Our findings could be used to assess the risks that current and next generation pesticides pose to honeybee populations.