In the hemipteran Rhodnius prolixus, successful post-prandial diuresis is accomplished through the synergistic actions of the peptidergic diuretic hormone RhoprCRF/DH and the biogenic amine 5-hydroxytryptamine (5-HT), and by an antidiuretic hormone RhoprCAPA-2 that terminates diuresis by inhibiting this synergy. Lateral neurosecretory cells (NSCs) in the mesothoracic ganglionic mass release RhoprCRF/DH, while midline NSCs release RhoprCAPA-2 during blood feeding. These NSCs co-express GPA2/GPB5, a conserved glycoprotein hormone involved in various physiological processes across bilaterians. This study investigates the influence of GPA2/GPB5 signaling on Malpighian tubule (MT) fluid secretion in R. prolixus. GPB5-like immunoreactivity in lateral and midline NSCs decreases following a blood meal, suggesting release and a role in diuresis. Downregulating the GPA2/GPB5 receptor LGR1 via RNA interference results in an increased basal fluid secretion rate in MTs, which is inhibited by the antidiuretic hormone RhoprCAPA-2. dsLGR1 treatment reduces the effects of RhoprCRF/DH and 5-HT on MT secretion and eliminates their synergism. RT-qPCR reveals that the transcript expression of the diuretic and antidiuretic hormone receptors are decreased in MTs of dsLGR1 injected insects, indicating that GPA2/GPB5 influences the expression of these other receptor transcripts. Downregulating LGR1 results in a smaller blood meal size and disrupts the normal time-course of diuresis. As LGR1 is the most abundantly expressed G protein-coupled receptor transcript in R. prolixus MTs, our results suggest that GPA2/GPB5 signaling has a critical role in regulating the timing and success of water retention in the unfed state, and in the complex processes associated with feeding and diuresis in R. prolixus.
