ABSTRACT

Neuroscience is one of the most prominent areas of modern biomedical research, but its impact on other fields, particularly those related to environmental and organismal biology, has been limited. This is because an increasing proportion of neuroscience research has been concentrated on just a few "model" animal species, such as the mouse and fruit fly, for which powerful genetic tools have been developed to manipulate their genomes. This focus stands in stark contrast to the early days of neuroscience research, in which a broader sampling of biodiversity led to important discoveries. Prominent examples include the discovery of how neurons become activated in the squid and how long-term memory is established in a marine slug. The primary goal of this proposal is to increase the diversity of animal species that can be used to advance neuroscience research by developing and applying modern neurogenetic tools for observing and manipulating neuronal activity in any animal species. As a proof-of-principle, state-of-the-art tools are developed for the honey bee and the American grasshopper. In addition to having immediate impact on basic neuroscience, the project has impact on applied research, as these two selected species are important pests and pollinators, respectively. The new tools and research findings from the proposed work is disseminated to the research community via NSF-funded initiatives to increase species diversity in genetic studies, and national and international workshops. The dissemination efforts include the development of an annual intensive summer course at Washington University that entails both lectures and hands-on experiences for organismal biologists who are interested in incorporating modern neuroscience and genetic tools into their research programs. Additional educational and training opportunities served by the project include public neuroscience outreach efforts in the St. Louis region, training of postdoctoral fellows, as well as mentoring of graduate, undergraduate, and high school students in hypothesis-driven neuroscience research.

The lack of species diversity in modern neuroscience research restricts the applicability and interpretations of general neurobiological principles in the context of organismal, ecological, and evolutionary questions. Therefore, the primary goal of this proposal is to increase animal species diversity in systems and behavioral neuroscience research by enabling easy adoption of universal genetic and transgenic tools for monitoring and manipulating neuronal activity in any animal species, with a specific emphasis on insects. The proposed approach is comprised of two steps. First, Cas9/CRISPR-dependent genome editing is 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 phiC31-integrase attP sites, enabling rapid screening of both white eye-color and RFP expression as markers of successful germline transformation. Second, the efficient phiC31-Integrase reaction is 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 Apis mellifera and the American grasshopper Schistocerca americana are 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. This NeuroTechnology Hub award is part of the BRAIN Initiative and NSF's Understanding the Brain activities.

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