Hybridization Chain Reaction RNA Whole-Mount Fluorescence In situ Hybridization of Chemosensory Genes in Mosquito Olfactory Appendages

Our group is interested in the chemo-sensory system of mosquitoes. We hope that by targeting a mosquito sense of taste and smell, we can stop it from finding us and biting us. Hybridization chain reaction fluorescent in situ hybridization is a recent development.

This technology is highly sensitive, robust, and capable of multiplexing several gene targets such that we can visualize the expression and localization of these genes simultaneously within their native environment. Special transcriptomics is a cutting edge method used in the field of spatial biology. This technology is capable of multiplexing several thousands of genes, and you can visualize the expression of these genes simultaneously.

Our protocol allows us to directly visualize which neurons in the antenna express any of the 50 or so possible ionotropic receptors. This provides us with a foundational roadmap to guide our understanding of how this classical factor receptors might mediate behaviors. Olfactory neurons within olfactory organs of an insect like the antenna, are typically buried inside a protective chitinous shell.

This makes visualizing olfactory receptor expression in those neurons very challenging. Our protocol uses a combination of chitinase treatments and a state-of-the-art fluorescence and see-through protocols to overcome these challenges.