Isolation functional characterization of allatotropin receptor from the cotton bollworm, Helicoverpa armigera

Insect allatotropin (AT) plays multi-functions including regulation of juvenile hormone synthesis, growth, development and reproduction. In the present study, the full-length cDNA encoding the AT receptor was cloned from the brain of Helicoverpa armigera (Helar-ATR). The ORF of Helar-ATR exhibited the characteristic seven transmembrane domains of the G protein-coupled receptor (GPCR) and was close to the ATR of Manduca sexta in the phylogenetic tree. The Helar-ATR expressed in vertebrate cell lines can be activated by Helar-AT and each Helar-ATL in a dose-responsive manner, in the following order: Helar-ATLI > Helar-ATLII > Helar-AT > Helar-ATLIII. Helar-ATLI and Helar-ATLII represented the functional ligands to Helar-ATR in vitro, while Helar-AT and Helar-ATLIII behaved as partial agonists. The in vitro functional analysis suggested that the Helar-ATR signal was mainly coupled with elevated levels of Ca²⁺ and independent of cAMP levels. Helar-ATR mRNA in larvae showed the highest level in the brain, followed by the thorax ganglion, abdomen ganglion, fat body and midgut. Helar-ATR mRNA levels in the complex of the brain-thoracic-abdomen ganglion on the 2nd day of the larval stage and during later pupal stages were observed to be relatively higher than in the wandering and early pupal stages.