The involvement of insulin/ToR signaling pathway in reproductive performance of Rhodnius prolixus.

Insulins are peptide hormones widely studied for their important regulatory roles in metabolism, growth and development. In insects, insulin signaling along with the target of rapamycin (ToR) are involved in detecting and interpreting nutrient levels. Recently, by transcriptome analysis we reported an up-regulation of transcripts involved in insulin/ToR signaling in unfed Rhodnius prolixus; however, this signaling pathway is only activated in fed insects. Here, continuing with the blood-gorging triatomine R. prolixus as a model, we report the direct effect of insulin/ToR signaling on reproductive performance. By immunofluorescence we identified cells in the brain with positive signal to the R. prolixus ILP (Rhopr-ILP1) and show that the insulin receptor and protein effectors downstream of insulin/ToR signaling activation, are differentially expressed in ovarian follicles dependent on their developmental stage. Using qPCR we find that the expression of transcripts involved in insulin signaling in the central nervous system (CNS), fat body and ovaries increase as the state of starvation progresses, promoting a more highly sensitized state to respond rapidly to ILP/IGF levels. In addition, using dsRNA injection and in vivo and ex vivo assays to promote signaling activation we demonstrate a direct participation of insulin/ToR signaling in coordinating the synthesis of the main yolk protein precursor, vitellogenin, thereby influencing the numbers of eggs laid per female. We thereby show a mechanism by which nutritional signaling regulates reproductive performance in a vector of Chagas disease. As reproduction is responsible for propagation of insect populations, this work is important for the development of innovative biocontrol methods.