Functional characterization of vitellogenin unveils novel roles in RHBP uptake and lifespan regulation in the insect vector Rhodnius prolixus

In insects, vitellogenesis plays a critical role in providing the energy reserves needed for embryonic development as it ensures the accumulation of yolk in the oocytes. Vitellogenin (Vg), the precursor to vitellin (Vt), is primarily synthesized in the fat body of females and transported to the oocytes via receptor-mediated endocytosis. In Rhodnius prolixus, a key vector of Chagas disease, two Vg genes, Vg1 and Vg2, were characterized. These genes share 65 % amino acid identity and present the conserved Vitellogenin_N, DUF1943, and VWD domains typical of Vg proteins across various species. We found that Vg1 is expressed at significantly higher levels than Vg2 in adult females. Still, the expression of both isoforms was also detected in organs such as the flight muscle, midgut, and ovary, as well as in males and nymphs. RNAi-mediated knockdown of Vg1 and Vg2 in adult females resulted in the production of yolk-depleted eggs with drastically reduced levels of Vg and RHBP, the second most import yolk protein in this species. Despite regular oviposition rates, most of these eggs were inviable, highlighting the essential role of Vg and RHBP in embryo development. Although Vg expression was detected in adult males, the mating of Vg-knockdown males with wild-type females did not impact oviposition or egg viability, indicating that male Vg is not crucial for oogenesis in this species. Interestingly, Vg knockdown increased lifespan for both males and females, suggesting additional physiological functions beyond reproduction. These findings reveal the importance of Vg in oogenesis and embryonic development in R. prolixus while also suggesting potential non-reproductive roles of Vg in adult insect physiology.