Allethrin is a widely used pyrethroid insecticide with an alkenylmethylcyclopentenolone group in its structure. We have analyzed its interaction with model and native membranes using DPH and its polar derivative TMA-DPH fluorescence polarization. Allethrin modified the bilayer order in the temperature range of the phase transition when incorporated into liposomes made with dimyristoyl-(DMPC), dipalmitoyl-(DPPC) and distearoyl-(DSPC) phosphatidylcholine. In DMPC: allethrin mixtures the pyrethroid decreased the bilayer order in the gel phase, without altering the liquid-crystalline one. In native membranes, DPH and TMA-DPH fluorescence polarization remained unchanged after incubation with allethrin. The release of hemoglobin was notably facilitated by the incorporation of allethrin into human erythrocytes. The results are discussed in terms of a possible aggregation of the insecticide in the lipid bilayer to create special domains with a consequent increase in membrane instability.