Oocyte membrane localization of vitellogenin receptor coincides with queen flying age, and receptor silencing by RNAi disrupts egg formation in fire ant virgin queens

In ant species in which mating flights are a strategic life-history trait for dispersal and reproduction, maturation of virgin queens occurs. However, the specific molecular mechanisms that mark this transition and the effectors that control pre-mating ovarian growth are unknown. The vitellogenin receptor (VgR) is responsible for vitellogenin uptake during egg formation in insects. In the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae), virgin queens have more abundant VgR transcripts than newly mated queens, but limited egg formation. To elucidate whether the transition to egg production involved changes in VgR expression, we investigated both virgin and mated queens. In both queens, western blot analysis showed an ovary-specific VgR band (about 202 kDa), and immunofluorescence analysis of ovaries detected differential VgR localization in early- and late-stage oocytes. However, the VgR signal was much lower in virgin queens ready to fly than in mated queens 8 h post mating flight. In virgin queens, the receptor signal was first observed at the oocyte membrane beginning at day 12 post emergence, coinciding with the 2 weeks of maturation required before a mating flight. Thus, the membrane localization of VgR appears to be a potential marker for queen mating readiness. Silencing of the receptor in virgin queens through RNA interference abolished egg formation, demonstrating that VgR is involved in fire ant ovary development pre mating. To our knowledge, this is the first report of RNA interference in any ant species and the first report of silencing of a hymenopteran VgR.