Acetylcholinesterase (AChE) is a key enzyme of the cholinergic nerve system. Of the two forms found in insects, the predominant one is active in the synapses and is the target of organophosphate and carbamate insecticides, while the role of the second is currently unknown. Two acetylcholinesterase cDNAs from the basal hexapod Orchesella villosa have been characterized and compared with others reported form insects. One form conforms well to the typical structure, while the other is characterized by an unusual 3′ region. No amino acid mutation could be directly associated with known resistance mutations in other insect species or to a clear signal of selection in the distribution of alleles, although the action of some population process is suggested.