Cloning and expression of long neuropeptide F and the role of FMRFamide-like peptides in regulating egg production in the Chagas vector, Rhodnius prolixus

• Long NPF has been cloned in Rhodnius (RhoprNPF) and is conserved across arthropods.
• RhoprNPF is predominantly expressed in the CNS and gut of instars and adults.
• 12 paired cells express RhoprNPF transcript in the brain of instars and adults.
• Adult female lateral oviducts contain cells that express RhoprNPF.
• RhoprNPF and FM/IRFamides increase egg production whereas MS and AST inhibit.

Long neuropeptide F (NPF) is a neuropeptide implicated in the control of feeding, digestion and reproduction in various insect species. Here we have isolated the cDNA sequence encoding NPF in Rhodnius prolixus (RhoprNPF). The RhoprNPF gene is composed of 3 exons and 2 introns, one of which is present in the peptide coding region. RhoprNPF is 42 amino acids long and has the characteristic RFamide C-terminus, which is common of FMRFamide-like peptides (FLPs). Quantitative PCR (qPCR) shows that RhoprNPF mRNA is present in higher amounts in fifth instars than in adults, implying that it may play a role in growth and development. In situ hybridization shows that the RhoprNPF transcript is present in median neurosecretory cells (MNSCs) in the brain, cells in the fifth instar hindgut and cells along the longitudinal muscle fibers of the adult female lateral oviducts. Injection of the last 8 amino acids of RhoprNPF (truncated RhoprNPF, AVAGRPRFa), which is considered to be the active core sequence for biological activity, into mated, fed, female adult R. prolixus decreased the number of eggs found in the ovaries as well as increased the number of eggs laid. This suggests that RhoprNPF may play a role in accelerating the process of ovulation from the ovary of the female R. prolixus. An increase in oogenesis was observed following the injection of other FLPs such as RhoprShortNPF, GNDNFMRFamide and AKDNFIRFamide, whereas the FLP, RhoprMS, and the allatostatin, RhoprAST-2, inhibited egg production.