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Contributed by Nancy Pierce on 12/06/2013
The BBB Research Scholarship is to support her research project, “In vivo tracking of CART neuropeptide dynamics in Danio rerio”. The results of the research will be presented at a future convention and may be published in BIOS. Research conducted under the supervision of Ian Woods.
CART (cocaine- and amphetamine-regulated transcript) is a neuropeptide with functions in behaviors such as food intake, body weight maintenance, and reward. The aim of this research project is to elucidate the in vivo dynamics of the expression, release, transport and receptor localization of the CART peptide. We plan to accomplish this through four main steps, using Split GFP as a reporter of CART localization. First, we will generate plasmid reagents that will enable an inducible cart-gfp11 (hs-gfp-11) and a ubiquitously-expressed gfp1-10 (actin-gfp1-10). Second, we will test the functionality of the hs-cart-gfp11 via established behavioral assays. Third, we will create transgenic zebrafish for the hs-cart-gfp11 and actin-gfp1-10 construct,confirm their fluorescence in transiently-transgenic larvae, and generate stable lines for these transgenes. Finally, we will generate reagents for genome editing via CRISPR or TALEN technologies to introduce the gfp11 label to the CART gene on its endogenous locus. Taken together, these experiments will elucidate the dynamics of CART signaling in the context of a living, functioning brain.