The primary goal of this project is to increase the diversity of animal species that can be used to advance neuroethological research by developing and applying modern neurogenetic tools for observing and manipulating neuronal activity in behaving animals. As a proof-of-principle, state-of-the-art tools will be developed for the European honey bee Apis mellifera, the Common eastern bumble bee Bombus impatiens, and the American bird grasshopper Schistocerca americana, which would have an immediate impact on basic and applied neuroscience research with these specific model organisms. The longer-term vision of the research team is to disseminate protocols, approaches, and molecular genetic tools to the broad research community to enable the application of advanced tools for studies of the neuronal basis to behavior in diverse species and various biological research areas.
Research Resources Being Developed and Disseminated
The initial goals of the project are to develop transgenic lines of insects for studying neuronal activity by using a two-steps process. First, Cas9/CRISPR-dependent genome editing will be used to replace the non-essential gene white with a DNA cassette that includes an eye-specific red fluorescent protein (RFP) flanked by two directional FC31-integrase attP sites, enabling rapid screening of both white eye-color and RFP expression as markers of successful germline transformation. Second, the efficient FC31-Integrasereaction will be used to replace the RFP cassette with a transgene of choice. As a proof-of-principle, transgenic lines that express the Ca2+reporter GCaMP6 in defined neuronal populations in the honey bee, the bumblebee, and the American grasshopper will be generated and tested for feasibility. By enabling the use of modern genetic tools to enhance neuroscience research in these two economically important insect species, which also serve as important models for basic organismal biological research, the proposed project is likely to have broad impact relevant to diverse research fields, including agriculture, neuroethology, animal behavior, pest ecology, and behavioral neuroscience.
Reaching Out to the Community
Research outcomes and assisted tool development will be communicated and implemented via a dedicated website and online tools, as well as via a short summer course.