20E-dependent tyrosine phosphorylation of phospholipase C gamma underpins egg development in the malaria vector Anopheles gambiae
The data set describes investigations using western blotting, immunofluorescence, RNA interference and the injection of pharmacological inhibitors to define the signalling events that underpin egg development in the malaria vector Anopheles gambiae following a blood meal. These events are of interest since a better understanding of how mosquitoes develop eggs may offer novel opportunities to intervene to reduce their reproductive output and thus malaria transmission. The data show that blood feeding is associated with a robust and tissue-specific increase in tyrosine phosphorylation in the reproductive tract and specifically in epithelial cells of the developing ovarian follicles. This tyrosine phosphorylation is inhibited by a tyrosine kinase inhibitor while also reducing the number of eggs that develop. We go on to show that one of the proteins phosphorylated after blood feeding is phospholipase C gamma and that this phosphorylation is also required for optimal egg development. We further link this phosphorylation event to the ecdysteroid hormone 20E which is produced by the female in response to blood feeding. We show that while 20E is necessary for PLC gamma phosphorylation it is not alone sufficient, suggesting the involvement of other blood feeding regulated factors such as insulin. Finally using both RNAi and pharmacological inhibitors we show that optimal phosphorylation of PLC gamma is dependent on signalling by Src family tyrosine kinases. Depletion or selective inhibition of these kinases reduces both phosphorylation of PLC gamma and the development of eggs. The bulk of the data deposited in this repository represent the original western blot and immunofluorescence images that were used as representative examples in the figures as well as those used to demonstrate the reproducibility of the observations and build the graphs presented.