Whole-mount in situ hybridization for analysis of gene expression during Aedes aegypti development

Abstract

Blood-feeding mosquitoes, including the dengue and yellow fever vector Aedes aegypti, transmit many of the world's deadliest diseases. Such diseases have resurged in developing countries and pose clear threats for epidemic outbreaks in developed countries. Recent mosquito genome projects have stimulated interest in the potential for arthropod-borne disease control by genetic manipulation of vector insects, and genes that regulate development are of particular interest. This protocol for whole-mount in situ hybridization can be used to analyze gene expression in Ae. aegypti embryos and larvae, a critical aspect of understanding developmental gene function in this vector mosquito.

Fig. 1. Gene expression in Ae. aegypti embryos

(A) fra and casein kinase (B, C) are expressed ventrally in the developing nerve cord at 55 hrs. of development. Lateral views of whole-mount embryos stained with the accompanying protocol are shown in A and B (anterior is oriented left). A filleted nerve cord is shown in C (anterior is oriented up).