Understanding the Molecular Mechanisms of Thermoreceptors - PROJECT SUMMARY/ABSTRACT Temperature significantly influences physiological processes and behaviors, impacting chemical reactions, biomolecule activity, and species distribution. Mosquitoes, vectors of diseases like malaria and dengue, are responsive to temperature cues, shaping survival, reproduction, and host-seeking behaviors. Fruit flies and mosquitoes utilize the same receptors for innocuous thermosensation, which can be studied in fruit flies due to the abundance of available genetic tools. Unraveling the cellular, molecular, and neural basis can offer insights into controlling vector-borne disease transmission. Drosophila melanogaster (fruit flies) is a model for studying insect thermosensory processes. Its thermosensory systems rely on manipulable sensory neurons and a balance between high- and low-temperature activated neurons. Specific neurons, including aristal heating and cooling cells, guide avoidance responses to temperature shifts. The warm receptor GR28B(D), which is essential for avoiding high temperatures, is not well understood. This proposal will uncover the precise molecular mechanism of GR28B(D) in sensing temperature changes. I will utilize the CRISPR-Cas9 system to determine critical protein/amino acid domains necessary for temperature sensing. Employing a top-down strategy, I will systematically identify the critical domains/amino acids associated with the receptor. The successful execution of this project will reveal significant insight into the molecular intricacies of temperature sensing in fruit flies. Our approach integrates genetic manipulations, electrophysiology, advanced imaging, and behavioral analysis to uncover specific roles of Exon 1 and smaller protein domains in GR28B(D), advancing our understanding of sensory processes in Drosophila melanogaster and related insects. The approaches employed in this proposal offer a breadth of innovative in vitro and in vivo techniques to the PI. Training for the expertise of these techniques will take place in the sponsor’s laboratory. Throughout the fellowship, career professional development training will occur through journal clubs, seminars, and associations at Virginia Tech, as well as at national and regional conferences to prepare the PI for a career as an independent researcher.
