Novel System for Long-Term Rearing and High-Throughput Assay of Zebrafish

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DESCRIPTION (provided by applicant): The overall goal of this proposed research is to develop a multi-well insert apparatus that will facilitate rapid automated detection of reporter genes in living fish from embryonic to juvenile stages of development. This system will be relevant for any zebrafish-based assay which utilizes fluorescent or luminescent reporter genes as a means of evaluating a biological question of interest. Zebrafish are becoming an important model for understanding human disease and for discovery of new therapeutic compounds. In particular, zebrafish are well-suited to large-scale screening procedures, providing an in vivo validation system that approaches high-throughput capacities. Full realization of this potential will require the development of automated assay platforms, such as reporter based systems utilizing fluorescent or luminescent read outs, and the inclusion of fish at more mature stages of development. During Phase I, components of a system will be developed that will enable embryos to be maintained in individual chambers for up to 1 month of development; and enable a high-throughput primary screen. During Phase II a second component of the system based on the PSI SideView imaging technology will be developed that will enable a more rigorous, image-based secondary screen. PUBLIC HEALTH RELEVANCE: This research will develop technologies that will enable the use of zebrafish models of human disease in high-throughput screens for active pharmaceutical compounds. One key area that this technology will benefit is regenerative medicine with a particular focus in this application on regeneration of insulin-producing pancreatic cells for diabetes treatment.