Identification and functional characterization of FGLamide-related allatostatin receptor in Rhodnius prolixus

Zandawala M, Orchard I

Insect Biochem. Mol. Biol. 2015 Feb;57:1-10

PMID: 25500190

Abstract

FGLamide-related ASTs (FGLa/ASTs) are a family of brain/gut peptides with numerous physiological roles, including inhibition of juvenile hormone (JH) biosynthesis by the corpora allata and inhibition of visceral muscle contraction. FGLa/ASTs mediate their effects by binding to a rhodopsin-like G-protein coupled receptor that is evolutionarily related to the vertebrate galanin receptor. Here we determine the cDNA sequence encoding FGLa/AST receptor (FGLa/AST-R) from the Chagas disease vector, Rhodnius prolixus (Rhopr-FGLa/AST-R), determine its spatial expression pattern using quantitative PCR and functionally characterize the receptor using a heterologous assay. Our expression analysis indicates that Rhopr-FGLa/AST-R is highly expressed in the central nervous system. The receptor is also expressed in various peripheral tissues including the dorsal vessel, midgut, hindgut and reproductive tissues of both males and females, suggesting a role in processes associated with feeding and reproduction. The possible involvement of Rhopr-FGLa/ASTs in the inhibition of JH biosynthesis is also implicated due to presence of the receptor transcript in the R. prolixus corpora cardiaca/corpora allata complex. The functional assay showed that various Rhopr-FGLa/ASTs activate the receptor, with EC50 values for the response in the nanomolar range. Moreover, Rhopr-FGLa/AST-R can couple with Gq alpha subunits and cause an increase in intracellular calcium concentration. Lastly, we tested various FGLa/AST analogs in our heterologous assay. These compounds also activated the receptor and thus have the potential to serve as insect growth regulators and aid in pest control.

Structure-activity relationships of two Rhodnius prolixus calcitonin-like diuretic hormone analogs

Zandawala M, Poulos C, Orchard I

Peptides 2014 Apr;

PMID: 24703964

Abstract

The calcitonin-like diuretic hormone (CT/DH) in Rhodnius prolixus influences various tissues associated with feeding-related physiological events. The receptors for this peptide have also been identified and shown to be expressed in these tissues. In the present study, we have investigated the effects of two R. prolixus CT/DH analogs (full-length form and N-terminal truncated form) on hindgut contractions and in a heterologous receptor expression system. The analogs contained the amino acid methyl-homoserine in place of methionine in order to prevent them from being oxidized and thus increase their stability. The full-length form of the analog retained all of its activity in our assays when compared to the endogenous peptide. Truncated analog displayed no activity in our assays.

Molecular characterization and possible biological roles of allatotropin in Rhodnius prolixus

Masood M, Orchard I

Peptides 2014 Mar;53:159-71

PMID: 24177575

Abstract

Allatotropins (ATs) are a family of neuropeptides that have been shown to stimulate the biosynthesis of juvenile hormone in certain insect species, and to have stimulatory activity on some visceral muscles. Here, we have examined the AT in Rhodnius prolixus. Molecular analysis revealed a cDNA fragment of 973 bp encoding one mature amidated AT (Rhopr-AT) with transcript levels observed in the central nervous system (CNS) and pool of fat body, trachea and associated peripheral nerves. AT-like immunoreactive neurons were found throughout the CNS and AT-like immunoreactive processes were present on some peripheral tissues. Bioassays based upon changes in hindgut and dorsal vessel contractions failed to demonstrate any myotropic effects of Rhopr-AT on these tissues; however Rhopr-AT stimulated contractions of muscles surrounding the salivary glands and secretion of saliva, as judged by the reduction in content of the cherry red saliva from the salivary glands. Serotonin stimulated an increase in peristaltic contractions of the gland though no secretion was observed. Co-application of Rhopr-AT and serotonin resulted in a more rapid secretion than either chemical alone.