Generation and analysis of transgenic zebrafish and mice for the study of Dlx function in the forebrain

Abstract

The vertebrate Dlx homeobox genes are organized as three convergently transcribed bigene clusters and show overlapping expression patterns in the forebrain, pharyngeal arches, sensory placodes and limb/fin buds. Little is known about how the Dlx genes are targeted to their sites of expression, or what particular roles individual genes play in development. Cis-acting regulatory elements have been identified in the intergenic and upstream regions of paired Dlx genes and are thought to play a major role in the Dlx regulation in the forebrain. My study was focused on two of the enhancer elements from the Dlxl/Dlx2 locus, I12b and URE2. We used these two elements to target reporter transgene expression to the zebrafish forebrain and analyzed their activity. We also successfully demonstrated the use of Fluorescent Activated Cell Sorting to isolate Dlx-Green Florescent Protein-expressing cells from the zebrafish forebrain. These results contribute to our understanding of Dlx regulation, function and evolution.