Corticotropin-releasing factor-like diuretic hormone 44 and five corresponding GPCRs in Drosophila suzukii: Structural and functional characterization

Diuretic hormones (DHs) activate corresponding G protein-coupled receptors (GPCRs), mediating the water and ion homeostasis in arthropods. There are two different DHs known to be expressed in insects, calcitonin (CT)-like DH31 and corticotropin-releasing factor (CRF)-like DH44. In this study, we identified and characterized a DH44 and five GPCR variants, DH44-R1 and DH44-R2a/b/c/d, in Drosophila suzukii (spotted-wing drosophila), causing detrimental damage to fresh and soft-skinned fruits. Among the five DH44 receptors, DH44-R1 was the longest GPCR and most strongly responded to DH44, and the other DH44-R2 splicing variants were relatively shorter and over 90 % similar to each other. Some DH44-Rs including DH44-R1 utilized both cAMP and Ca²⁺ as second messengers. Interestingly, DH44-R1 was dominantly expressed in the brain, whereas DH44-R2 variants were dominant in the digestive organs, particularly the Malpighian tubules (MTs) by their gene expressions. The results suggest that DH44 may have multiple physiological functions, including the regulation of the sleep-wake cycle and diuretic activity. Injection of DH44 stimulated fluid secretion in adults, and the rate of the excretion increased in a dose-dependent manner. Moreover, when the flies were injected with a mixture of DH31 and DH44, a high mortality rate was observed. Here, we demonstrate the gene structures, expressions, characterization of DH44 and five GPCRs, their second messengers, and the effects of DH peptides on the fly. These investigations offer molecular insights into the physiological roles of the DH system and may assist in the fundamental aspects of developing D. suzukii management in the field.