Identification of a tachykinin receptor and its implication in carbohydrate and lipid homeostasis in Rhodnius prolixus, a chagas disease vector.

Neuropeptides and their receptors are fundamentally important in regulating many physiological and behavioural processes in insects. In this work, we have identified, cloned, and sequenced the tachykinin receptor (Rhopr-TKR) from Rhodnius prolixus, a vector of Chagas disease. The receptor is a G protein-coupled receptor belonging to the Rhodopsin Family A. The total length of the open reading frame of the Rhopr-TKR transcript is 1110 bp, which translates into a receptor of 338 amino acids. Fluorescent in-situ RNA-hybridization (FISH) for the Rhopr-TKR transcript shows a signal in a group of six bilaterally paired neurons in the protocerebrum of the brain, localized in a similar region as the insulin producing cells. To examine the role of tachykinin signaling in lipid and carbohydrate homeostasis we used RNA interference. Downregulation of the Rhopr-TKR transcript led to a decrease in the size of blood meal consumed and a significant increase in circulating carbohydrate and lipid levels. Further investigation revealed a close relationship between tachykinin and insulin signaling since the downregulation of the Rhopr-TKR transcript negatively affected the transcript expression for insulin-like peptide 1 (Rhopr-ILP1), insulin-like growth factor (Rhopr-IGF) and insulin receptor 1 (Rhopr-InR1) in both the central nervous system and fat body. Taken together, these findings suggest that tachykinin signaling regulates lipid and carbohydrate homeostasis via the insulin signaling pathway.

Expression and functional characterization of tachykinin-related peptides in the blood-feeding bug, Rhodnius prolixus.

Tachykinins (tachykinin-related peptides, TRPs) are multifunctional neuropeptides that have widespread distribution in the central nervous system (CNS) and in the gastrointestinal tract of many insects, and most have been shown to stimulate contractions of visceral muscles. Invertebrate TRPs carry a characteristic conserved C-terminal pentapeptide (FXGXR-amide) and most of them share some amino acid sequence similarities (approx. 45%) with the vertebrate and mammalian tachykinin family. We have functionally characterized the tachykinins in R. prolixus (Rhopr-TKs) and partially cloned the transcript that encodes for the peptide precursor. The transcript encodes 8 Rhopr-TKs, 7 of which are unique with Rhopr-TK 5 having 2 copies. The spatial distribution analysis of the Rhopr-TK transcript indicates that the highest expression levels are in the CNS, but transcript expression is also associated with salivary glands, fat body, dorsal vessel, and the various gut compartments. Rhopr-TK 1, 2 and 5 significantly increase the frequency and amplitude of peristaltic contractions of the salivary glands. Hindgut muscle also displayed a dose-dependent increase in basal tonus in response to Rhopr-TK1, 2 and 5. TK-like immunoreactivity was seen in a small group of processes that are situated on the lateral margins of the hindgut. Interestingly, kinin-like immunoreactivity is seen in immunoreactive processes on the lateral margin of the hindgut as well as fine processes covering the entire hindgut. Co-localization studies show that TK-like staining is always co-localized with kinin-like immunoreactivity, whereas kinin-like staining is seen in the fine processes that are devoid of TK-like immunoreactivity indicating that TKs are most likely released together with kinins to act on the hindgut. Rhopr-Kinin 2 is a potent stimulator of hindgut muscle contraction in R. prolixus. Addition of Rhopr-Kinin 2 and Rhopr-TK 2 to the hindgut leads to a contraction that was additive of the effects of Rhopr-Kinin 2 and Rhopr-TK 2 alone.