An system for investigation of invasion of the salivary gland of mosquitoes.

sporozoites associated with the midgut and in the hemolymph of mosquitoes differ from sporozoites in the secretory cavities and ducts of the insects' salivary glands in their transcriptome, proteome, motility, and infectivity. Using an salivary gland culture system incorporating simple microfluidics and transgenic with the fluorescent protein gene under the transcriptional control of the promoter whose expression served as a proxy for parasite maturation, we observed rapid parasite maturation in the absence of salivary gland invasion. While expression was only detectable in sporozoites within the salivary glands (mature parasites) as expected, the simple exposure of sporozoites to dissected salivary glands led to rapid parasite maturation as indicated by mCherry expression. These results suggest that previous efforts to develop and systems for investigating sporozoite interactions with mosquito salivary glands have likely been unsuccessful in part because the maturation of sporozoites leads to a loss in the ability to invade salivary glands.