Determining the Basic Biology Underlying the Growth of Wolbachia Inside Infected Female Mosquitoes’ Ovaries
Wolbachia is a commonly found bacterium in arthropods and nematodes and can induce Cytoplasmic Incompatibility (CI) in mosquitoes' embryos. With the recent outbreak of Zika and malaria, CI is being reinvestigated as a tool to control the transmission of mosquito-transmitted diseases (Hoffmann et al., 2011; Walker et al., 2011). With the time when Wolbachia peaks in mosquitoes’ ovaries remains unknown, we hypothesized that unfed mosquitoes’ ovaries have the lowest Wolbachia number compared to those fed with bloodmeal. First, we extract the DNA from dissected ovaries from fed mosquitoes and measure Wolbachia abundance by PCR. This established a baseline for the next experiments. Then, we treated our infected mosquito larvae with tetracycline, which has already been shown to eliminate all Wolbachia in infected mosquitoes and determine Wolbachia levels in mosquito ovaries after each generation of treatment (Beckmann and Fallon, 2012). We also looked at the proteins in the ovary at each generation using SDS-PAGE to see if some bands disappear. Our protein gel results showed that there are some proteins in normal ovaries that did not show up in Wolbachia-infected ovaries. Our next step is to identify the proteins using mass spectrometry and IEF-PAGE.