Molecular response of Drosophila melanogaster tyramine receptor cascade to plant essential oils

Abstract

This paper reports the role of the tyramine (TA) receptor cascade in the insecticidal activity of plant essential oils. A TA receptor cDNA encoding a putative seven transmembrane domain G-protein coupled receptor was amplified from Drosophila melanogaster head cDNA phage library. The encoded protein contains 601 amino acids and has a sequence similar to other biogenic amine receptors. This protein was expressed in Drosophila S2 cells for radioligand binding studies with the ligand 3H-TA. Competitive binding studies comparing biogenic amines that could potentially function as endogenous ligands have demonstrated that this receptor had the highest affinity for TA ( K i = 1.27 μ M ) followed by DL-octopamine, dopamine, serotonin and histamine. TA decreased the forskolin-increased cAMP levels (IC 50=5.802 μM) and increased [Ca 2+] i through the receptor expressed in S2 cells. The toxicity rank order of the tested plant essential oils against wild type D. melanogaster fly demonstrated a pattern similar to their effect on receptor binding activity and changes in cAMP level and [Ca 2+] i . The toxicity of two of these chemicals was eliminated when tested against the TA receptor mutant (TyrR neo30) Drosophila strain. Therefore, the data indicates a correlation between cellular changes and insecticidal activity of tested plant essential oils, and suggests that the toxicity of at least two of these chemicals is mediated through the TA receptor.