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Finetti, Luca; Ferrari, Federica; Caló, Girolamo; Cassanelli, Stefano; De Bastiani, Morena; Civolani, Stefano; Bernacchia, Giovanni
Pesticide biochemistry and physiology, 20/May , Letnik: 165Journal Article
This study proposes a biochemical and molecular model for the interaction between the Drosophila suzukii type 1 tyramine receptor (DsTAR1) and monoterpenes. A preliminary molecular and functional characterization of DsTAR1 cDNA revealed that a 1.8 kb long ORF codes for a 600 amino acid polypeptide featuring seven transmembrane domains, as expected for a GPCR. A stable HEK 293 cell line expressing DsTAR1 was tested for responsiveness to tyramine (TA) and octopamine (OA). In intracellular calcium mobilization studies, TA led to a concentration-dependent increase in Ca2+i (pEC50 ~ 6.40), completely abolished by pre-incubation with the antagonist yohimbine 1 μM. Besides, in dynamic mass redistribution (DMR) studies, TA evoked a positive DMR signal in a concentration-dependent manner (pEC50 ~ 6.80). The recombinant cell line was then used to test three monoterpenes (thymol, carvacrol and α-terpineol) as putative ligands for DsTAR1. The terpenoids showed no agonist effects in both DMR and calcium mobilization assays, but they increased the potency of the endogenous ligand, TA, acting as positive allosteric modulators. Moreover, expression analysis on adults D. suzukii, exposed for 24, 72 or 120 h to a sublethal concentration of the three monoterpenes, showed a downregulation of DsTAR1. This evidence has led to hypothesize that the downregulation of DsTAR1 might be a compensatory mechanism in response to the positive allosteric modulation of the receptor induced by monoterpenes. Therefore, these findings might be useful for the development of a new generation of biopesticides against Drosophila suzukii, targeting TAR1. Display omitted •DsTAR1 is a GPCR expressed at high levels in the head of male and female flies.•DsTAR1 is dose-dependently activated by TA and selectively blocked by yohimbine.•Monoterpenes modulate DsTAR1 enhancing the TA response•Monoterpenes exposure induce a downregulation of DsTAR1 in D. suzukii•The monoterpene-enhanced TA response affect mRNA levels in a compensatory fashion.
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