Transcriptome analysis of Aedes albopictus midguts infected by dengue virus identifies a gene network module highly associated with temperature.

BACKGROUND

Dengue is prevalent worldwide and is transmitted by Aedes mosquitoes. Temperature is a strong driver of dengue transmission. However, little is known about the underlying mechanisms.

METHODS

Aedes albopictus mosquitoes exposed or not exposed to dengue virus serotype 2 (DENV-2) were reared at 23 °C, 28 °C and 32 °C, and midguts and residual tissues were evaluated at 7 days after infection. RNA sequencing of midgut pools from the control group, midgut breakthrough group and midgut nonbreakthrough group at different temperatures was performed. The transcriptomic profiles were analyzed using the R package, followed by weighted gene correlation network analysis (WGCNA) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis to identify the important molecular mechanisms regulated by temperature.

RESULTS

The midgut infection rate and midgut breakthrough rate at 28 °C and 32 °C were significantly higher than those at 23 °C, which indicates that high temperature facilitates DENV-2 breakthrough in the Ae. albopictus midgut. Transcriptome sequencing was performed to investigate the antiviral mechanism in the midgut. The midgut gene expression datasets clustered with respect to temperature, blood-feeding and midgut breakthrough. Over 1500 differentially expressed genes were identified by pairwise comparisons of midguts at different temperatures. To assess key molecules regulated by temperature, we used WGCNA, which identified 28 modules of coexpressed genes; the ME3 module correlated with temperature. KEGG analysis indicated that RNA degradation, Toll and immunodeficiency factor signaling and other pathways are regulated by temperature.

CONCLUSIONS

Temperature affects the infection and breakthrough of Ae. albopictus midguts invaded by DENV-2, and Ae. albopictus midgut transcriptomes change with temperature. The candidate genes and key pathways regulated by temperature provide targets for the prevention and control of dengue.