Aedes aegypti mosquito eggshell proteins in response to RNAi-EOF1

Aedes aegypti mosquito ovarian follicles develop synchronously after taking a blood meal. A single layer of follicular epithelial cells surrounding the oocyte is responsible for secreting a majority of eggshell structural components between 18 and 54 hours post blood meal. We previously identified a protein called eggshell organizing factor 1 (EOF1). When we knocked down EOF1 with RNA interference in female adult mosquitoes, they laid eggs that were not properly melanized, and fragile eggs did not contain viable embryos. Here, we aimed to determine putative downstream eggshell proteins in RNAi-EOF1 female mosquitoes. Our eggshell proteomics identified 220 proteins, suggesting that mosquito extracellular eggshells are composed of a complex mixture of proteins. These proteins are involved in the formation of an intact eggshell that protects the embryonic development and larva from the environment for long periods of time.